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Alam, Nur, I. Made Budiarsa, and Dewi Tureni. "Prediksi Struktur Tiga Dimensi Protein β-NGF (Nerve Growth Factor) Burung Merpati (Columba livia)." JURNAL ILMIAH SAINS 20, no. 2 (September 13, 2020): 106. http://dx.doi.org/10.35799/jis.20.2.2020.28857.
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Protein β-NGF berperan dalam kelangsungan hidup sel neuron dan diferensiasi embrio aves yang diekspresikan oleh gen NGF. Informasi protein β-NGF telah diidentifikasi pada ayam dan masih terbatas dilaporkan pada merpati. Tujuan penelitian adalah memprediksi struktur tiga dimensi protein β-NGF merpati untuk menambah informasi karakter proteinnya. Protein target diperoleh dari server UniProt dengan kode akses B4ZE95. Prediksi struktur tiga dimensi dan karakterisasi protein β-NGF menggunakan server SWISS-MODEL dan program Chimera. Hasil analisis menunjukkan bahwa protein β-NGF merpati dan template memiliki nilai identity sebesar 89,91%, QMEAN 0,51 serta GMQE sebesar 0,44. Struktur tiga dimensi protein B4ZE95 memiliki asam amino hidrofobik (Ile, Leu, Trp, Phe, Val, Ala, Gly, Met, Cys); karboksil (Glu, Ser, Asp); amina (Lys, Thr) dengan 83 ikatan hidrogen.Kata kunci: columba livia, chimera, protein β-NGF, swiss-model Three Dimensional Protein Structure Prediction of β-NGF(Nerve Growth Factor) on Pigeon (Columba livia)ABSTRACTThe β-NGF protein plays a role in the survival of neuron cells and differentiation of avian embryo which is expressed by NGF gene. Information of β -NGF protein was identified in chicken and it is still limited reported in pigeon. The purpose of this study is to predict the three-dimensional structure of pigeon β-NGF protein to add information about its protein character. The target protein is obtained from UniProt server with access code B4ZE95. The prediction of three-dimensional structure and characterization of β-NGF protein were carried out using SWISS-MODEL server and Chimera program. The analysis showed that pigeon β-NGF protein and template had identity of 89,91%, QMEAN 0,51 and GMQE of 0,44. The three-dimensional structure of the B4ZE95 protein has hydrophobic amino acids namely (Ile, Leu, Trp, Phe, Val, Ala, Gly, Met, Cys); carboxyl namely (Glu, Ser, Asp); amine namely (Lys, Thr) with 83 hydrogen bonds.Keywords: columba livia, chimera, protein β-NGF, swiss model
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Zhao, Guang-Hua, Ping Yu, Xiao-Song Hu, and Lei Zhao. "Effect of Zn(II) on the Structure and Biological Activity of Natural β-NGF." Acta Biochimica et Biophysica Sinica 36, no. 2 (February 1, 2004): 99–104. http://dx.doi.org/10.1093/abbs/36.2.99.
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Abstract Only β-NGF, the subunit of the 7S NGF complex, exhibits NGF activity, but the function of the zinc ion in native β-NGF has received little attention. Flameless atomic absorption spectroscopy (FAAS) measurements reveal that native β-NGF contains Zn(II) with a Zn(II)/β-NGF stoichiometry of 1:14.6. The presence of Zn(II) in the native molecule results in significant changes of the secondary structure and local tertiary structure around Trp(s) with respect to those of apo β-NGF, as suggested by spectra of fluorescence and circular dichrosim. Stopped-flow studies show that there are at least two steps during the interaction of Zn(II) with the apo form. In comparison with its apo form, the native β-NGF shows a higher ability to trigger the proliferation of TF1 cells and mediate the survival of PC12. Thus it is most likely that the structural changes caused by the presence of Zn(II) directly lead to the increase in the biological activity of β-NGF. All results indicate that Zn(II) in native β-NGF plays an important role in the structure and the biological activity of the protein.
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Montagnoli, Claudia, Roberto Tiribuzi, Lucia Crispoltoni, Alessandra Pistilli, Anna Maria Stabile, Francesco Manfreda, Giacomo Placella, Mario Rende, and Giuliano G. Cerulli. "β-NGF and β-NGF receptor upregulation in blood and synovial fluid in osteoarthritis." Biological Chemistry 398, no. 9 (August 28, 2017): 1045–54. http://dx.doi.org/10.1515/hsz-2016-0280.
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Abstract Osteoarthritis (OA) of the knee is the most common form of non-traumatic joint disease. Previous studies have shown the involvement of β-NGF and its receptors TrKA and p75NTR in OA-related pain, but their role in its pathogenesis is still unclear. The aim of our study was to investigate the amount of β-NGF and the expression levels of its receptors on cells isolated from synovial fluid and blood from OA patients who had undergone total knee arthroplasty, in order to check any possible correlation with the disease staging. Our results show a progressive stage-related increase of β-NGF and its receptors both in serum and synovial fluid. Furthermore, with respect to control subjects, OA patients show an increased amount of inflammatory monocytes along with an increased expression of β-NGF, TrKA and p75NTR. In conclusion, our study suggests a stage-related modulation of β-NGF and its receptors in the inflammatory process of OA.
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Stuart, C. C., J. L. Vaughan, C. M. Kershaw-Young, J. Wilkinson, R. Bathgate, and S. P. de Graaf. "Effects of varying doses of β-nerve growth factor on the timing of ovulation, plasma progesterone concentration and corpus luteum size in female alpacas (Vicugna pacos)." Reproduction, Fertility and Development 27, no. 8 (2015): 1181. http://dx.doi.org/10.1071/rd14037.
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Ovulation in camelids is induced by the seminal plasma protein ovulation-inducing factor (OIF), recently identified as β-nerve growth factor (β-NGF). The present study measured the total protein concentration in alpaca seminal plasma using a bicinchoninic acid (BCA) protein quantification assay and found it to be 22.2 ± 2.0 mg mL–1. To measure the effects of varying doses of β-NGF on the incidence and timing of ovulation, corpus luteum (CL) size and plasma progesterone concentration, 24 female alpacas were synchronised and treated with either: (1) 1 mL 0.9% saline (n = 5); (2) 4 µg buserelin (n = 5); (3) 1 mg β-NGF protein (n = 5); (4) 0.1 mg β-NGF (n = 5); or (5) 0.01 mg β-NGF (n = 4). Females were examined by transrectal ultrasonography at 1–2-h intervals between 20 and 45 h after treatment or until ovulation occurred, as well as on Day 8 to observe the size of the CL, at which time blood was collected to measure plasma progesterone concentrations. Ovulation was detected in 0/5, 5/5, 5/5, 3/5 and 0/4 female alpacas treated with saline, buserelin, 1, 0.1 and 0.01 mg β-NGF, respectively. Mean ovulation interval (P = 0.76), CL diameter (P = 0.96) and plasma progesterone concentration (P = 0.96) did not differ between treatments. Mean ovulation interval overall was 26.2 ± 1.0 h. In conclusion, buserelin and 1 mg β-NGF are equally effective at inducing ovulation in female alpacas, but at doses ≤0.1 mg, β-NGF is not a reliable method for the induction of ovulation.
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Goins, William F., Kevin A. Lee, James D. Cavalcoli, Mark E. O’Malley, Steven T. DeKosky, David J. Fink, and Joseph C. Glorioso. "Herpes Simplex Virus Type 1 Vector-Mediated Expression of Nerve Growth Factor Protects Dorsal Root Ganglion Neurons from Peroxide Toxicity." Journal of Virology 73, no. 1 (January 1, 1999): 519–32. http://dx.doi.org/10.1128/jvi.73.1.519-532.1999.
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ABSTRACT Nerve growth factor β subunit (β-NGF) transgene delivery and expression by herpes simplex virus type 1 (HSV-1) vectors was examined in a cell culture model of neuroprotection from hydrogen peroxide toxicity. Replication-competent (tk− K mutant background) and replication-defective (ICP4−;tk− S mutant background) vectors were engineered to contain the murine β-NGF cDNA under transcriptional control of either the human cytomegalovirus immediate-early gene promoter (HCMV IEp) (e.g., KHN and SHN) or the latency-active promoter 2 (LAP2) (e.g., KLN and SLN) within the viral thymidine kinase (tk) locus. Infection of rat B103 and mouse N2A neuronal cell lines, 9L rat glioma cells, and Vero cells with the KHN or SHN vectors resulted in the production of β-NGF-specific transcripts and β-NGF protein reaching a maximum at 3 days postinfection (p.i.). NGF protein was released into the culture media in amounts ranging from 10.83 to 352.86 ng/ml, with the highest levels being achieved in B103 cells, and was capable of inducing neurite sprouting of PC-12 cells. The same vectors produced high levels of NGF in primary dorsal root ganglion (DRG) cultures at 3 days. In contrast to HCMV IEp-mediated expression, the LAP2-NGF vectors showed robust expression in primary DRG neurons at 14 days. The neuroprotective effect of vector produced NGF was assessed by its ability to inhibit hydrogen peroxide-induced neuron toxicity in primary DRG cultures. Consistent with the kinetics of vector-mediated NGF expression, HCMV-NGF vectors were effective in abrogating the toxic effects of peroxide at 3 but not 14 days p.i. whereas LAP2-NGF vector transduction inhibited apoptosis in DRG neurons at 14 days p.i. but was ineffective at 3 days p.i. Similar kinetics of NGF expression were observed with the KHN and KLN vectors in latently infected mouse trigeminal ganglia, where high levels of β-NGF protein expression were detected at 4 wks p.i. only from the LAP2; HCMV-NGF-driven expression peaked at 3 days but could not be detected during HSV latency at 4 weeks. Together, these results indicate that (i) NGF vector-infected cells produce and secrete mature, biologically active β-NGF; (ii) vector-synthesized NGF was capable of blocking peroxide-induced apoptosis in primary DRG cultures; and (iii) the HCMV-IEp functioned to produce high levels of NGF for several days; but (iv) only the native LAP2 was capable of long-term expression of a therapeutic gene product in latently infected neurons in vivo.
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Dünker, Nicole, Norbert Schuster, and Kerstin Krieglstein. "TGF-β modulates programmed cell death in the retina of the developing chick embryo." Development 128, no. 11 (June 1, 2001): 1933–42. http://dx.doi.org/10.1242/dev.128.11.1933.
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Programmed cell death (PCD) is a key phenomenon in the regulation of cell number in multicellular organisms. We have shown that reduction of endogenous transforming growth factor β (TGF-β) prevents apoptotic PCD of neurons in the developing peripheral and central nervous system, suggesting that TGF-β is an important mediator of ontogenetic neuron death. Previous studies suggested that there are other pro-apoptotic molecules, nerve growth factor (NGF) and brain-derived neurotrophic factor, that induce cell death in the nervous system. In the developing chick retina, NGF induces PCD by activation of the p75 receptor. We have studied the role of TGF-β and its putative interdependence with NGF-mediated PCD in the chick retina. We found that TGF-β is present in the developing chick retina during the period of PCD and is essentially required to regulate PCD of retinal cells. TGF-β2, TGF-β3 and the ligand-binding TGF-β receptor can be detected immunocytochemically in the central retina, a region where apoptosis is most prominent during the early period of PCD. Application of a TGF-β-neutralizing antibody to chick embryos in ovo resulted in a decrease in the number of TUNEL-positive cells and a reduction of free nucleosome levels. In terms of magnitude, reduction of PCD caused by the neutralization of endogenous TGF-β was equivalent to that seen after anti-NGF application. Neutralization of both factors did not result in a further decrease in apoptosis, indicating that NGF and TGF-β may act on the same cell population. Furthermore, neutralization of TGF-β did not affect the expression of NGF or the p75-receptor. Our results suggest that TGF-β and NGF are both required to regulate cell death in the chick retina in vivo.
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Wang, Fei, Kun Yuan, Xiaogang Zhou, Dawei Xu, Yuyu Sun, Feihu Chen, and Peiji Wang. "Nerve Growth Factors Promotes Osteogenic Differentiation Through TGF-β and BMP-9 Signaling Pathways in Bone Mesenchymal Stem Cells." Journal of Biomaterials and Tissue Engineering 10, no. 1 (January 1, 2020): 46–52. http://dx.doi.org/10.1166/jbt.2020.2199.
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Bone mesenchymal stem cells (BMSCs) are related to fracture healing. In this paper, we study the effect of Nerve growth factors (NGF) on osteogenic differentiation of BMSCs and related mechanisms. The results indicated that NGF has a dose-dependent promotion effect on osteogenic differentiation of BMSCs, ALP activity, and protein expression of OPN, Osterix, Runx2, COL1A1. Further investigation showed that NGF enhanced the expression of TGF-β and BMP9, and increased phosphorylation of Smad and p38 MAPK. Moreover, block of BMP and TGF-β pathway reduced the promoted effect of NGF on osteogenic differentiation. The data revealed that NGF increased osteogenic differentiation through increased phosphorylation of Smad and MAPK p38 by regulating TGF-β and BMP pathway in BMSCs. Thus, NGF may be used in bone growth and fracture healing by improving osteogenic differentiation of BMSCs.
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Koga, Takeru, Takaiku Sakamoto, Eiji Sakuradani, and Akihiro Tai. "Neurite Outgrowth-Promoting Activity of Compounds in PC12 Cells from Sunflower Seeds." Molecules 25, no. 20 (October 16, 2020): 4748. http://dx.doi.org/10.3390/molecules25204748.
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In the current super-aging society, the establishment of methods for prevention and treatment of Alzheimer’s disease (AD) is an urgent task. One of the causes of AD is thought to be a decrease in the revel of nerve growth factor (NGF) in the brain. Compounds showing NGF-mimicking activity and NGF-enhancing activity have been examined as possible agents for improving symptoms. In the present study, sunflower seed extract was found to have neurite outgrowth-promoting activity, which is an NGF-enhancing activity, in PC12 cells. To investigate neurite outgrowth-promoting compounds from sunflower seed extract, bioassay-guided purification was carried out. The purified active fraction was obtained by liquid-liquid partition followed by some column chromatographies. Proton nuclear magnetic resonance and gas chromatography-mass spectrometry analyses of the purified active fraction indicated that the fraction was a mixture of β-sitosterol, stigmasterol and campesterol, with β-sitosterol being the main component. Neurite outgrowth-promoting activities of β-sitosterol, stigmasterol, campesterol and cholesterol were evaluated in PC12 cells. β-Sitosterol and stigmasterol showed the strongest activity of the four sterol compounds (β-sitosterol ≈ stigmasterol > campesterol > cholesterol), and cholesterol did not show any activity. The results indicated that β-sitosterol was the major component responsible for the neurite outgrowth-promoting activity of sunflower seeds. Results of immunostaining also showed that promotion by β-sitosterol of neurite formation induced by NGF was accompanied by neurofilament expression. β-Sitosterol, which showed NGF-enhancing activity, might be a candidate ingredient in food for prevention of AD.
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Selvaraju, Vaithinathan, Jeganathan R. Babu, and Thangiah Geetha. "Salivary Neurotrophins Brain-Derived Neurotrophic Factor and Nerve Growth Factor Associated with Childhood Obesity: A Multiplex Magnetic Luminescence Analysis." Diagnostics 12, no. 5 (May 3, 2022): 1130. http://dx.doi.org/10.3390/diagnostics12051130.
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Obesity is linked with higher inflammatory markers and is characterized by chronic low-grade inflammation. Neurotrophins brain-derived neurotrophic factor (BDNF) and β-nerve growth factor (β-NGF), in addition to their neuronal functions, act on several immune cells and have been recently designated as metabokines due to their regulatory role in energy homeostasis and food intake. The current study evaluates the salivary BDNF and β-NGF and their association with anthropometric measurement, blood pressure, and salivary insulin in children. Anthropometric measurements and saliva samples were obtained from 76 children, aged 6–10 years. Multiplex analysis was carried out for the salivary analysis of BDNF, NGF, and insulin by human magnetic Luminex performance assay. Statistical analysis was performed to analyze the best fit diagnostic value for biomarkers and the relationship of the neurotrophic levels of BDNF and NGF with obesity measures and blood pressure. Salivary BDNF and β-NGF showed a significantly higher concentration in obese children than normal-weight children. Both neurotrophins are positively associated with obesity anthropometric measures, blood pressure, and salivary insulin. Multinominal regression analysis reported a significant association between salivary BDNF, β-NGF, insulin, and systolic pressure adjusted for age, gender, income, and maternal education. The salivary concentration of BDNF and NGF was higher in obese children, and it is positively associated with anthropometric measures, suggesting that neurotrophins can be used as a non-invasive predictor of obesity-related complications in children.
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Masdeu, M., R. M. Garcia-Garcia, R. Cardinali, P. Millan, M. Arias Alvarez, C. Castellini, P. L. Lorenzo, and P. G. Rebollar. "271 INDUCTION OF OVULATION IN RABBIT DOES USING PURIFIED NERVE GROWTH FACTOR AND CAMEL SEMINAL PLASMA." Reproduction, Fertility and Development 27, no. 1 (2015): 224. http://dx.doi.org/10.1071/rdv27n1ab271.
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The presence of an ovulation-inducing factor (OIF) in the seminal plasma (SP) of several species with spontaneous and induced ovulation, including the rabbit, has been documented. Recent studies have demonstrated that the OIF in the SP of camels (SPCAM) is a nerve growth factor (β-NGF). The aim of this study was to determine if purified β-NGF from mouse submandibular glands or SPCAM could provoke ovulation induction in the rabbit doe. A total of 35 females were synchronized with 25 IU of equine chorionic gonadotropin (Serigan, Laboratorios Ovejero, Spain) and allocated into 4 groups. Forty-eight hours later (Day 0), does were given a single dose (IM) of 1 mL of saline solution (SS; n = 8); 1 mL of gonadorelin (GnRH; Inducel, Laboratorios Ovejero, Spain; n = 9); 24 µg of β-NGF (2.5S-NGF; Promega, USA; n = 10); or 1 mL of centrifuged raw camel SP (SPCAM; 127 pg mL–1 NGF; n = 8). After treatment, an empty catheter was introduced through the vagina to simulate the nervous/mechanical stimulus of coitus (4 animals per group). Plasma LH concentrations were determined in blood samples taken 30 min before treatment and at 0, 30, 60, 90, and 120 min after injection. Progesterone concentrations were assessed at 0 and 120 min and every 2 days until Day 6 after treatment. Concentrations of β-NGF in camel SP and hormone determinations were made by enzyme immunoassay. Ovulation rate (OR) was determined after euthanasia on Day 7. Statistical analyses using CATMOD and MIXED procedures of the SAS program to compare OR data and hormone concentrations, respectively, were performed. Ovulation occurred in 100% of GnRH (9/9), 33% (3/10) of NGF, 25% (2/8) of SS, and 0% (0/8) of SPCAM groups. Both NGF and SS ovulated females had significantly lower LH concentration than GnRH group throughout all preovulatory surge (P < 0.001). When does were not stimulated with catheter introduction, only those from the GnRH (5/5) and NGF (1/6) groups ovulated. Total number of corpora lutea in ovulated does was similar (15.9 ± 1.9, 17.0 ± 4.2, and 14.3 ± 3.1 CL in GnRH, SS, and NGF groups, respectively). Plasma P4 concentrations were normally increased from Day 2 to 6 in ovulated rabbits of all groups, but were lower at 120 min (P < 0.001) for the NGF and SS does, reaching similar levels than GnRH does at 6 days post-treatment. In the present study, β-NGF from mouse submandibular glands, but not from raw camel SP, induced ovulation in rabbit females, independently of nervous stimulus. Nonetheless, the possible low dose of β-NGF used and the origin could have been responsible for the lack of a more acute effect.We acknowledge CM, FSE, and AGL2011-23822 for funding.
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Borsani, G., A. Pizzuti, E. I. Rugarli, A. Falini, V. Silani, A. Sidoli, G. Scarlato, and F. E. Baralle. "cDNA sequence of human β-NGF." Nucleic Acids Research 18, no. 13 (1990): 4020. http://dx.doi.org/10.1093/nar/18.13.4020.
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Che, Xin, Zhenjie Wang, Yining Liu, Yanhui Sun, and Hongzhuo Liu. "Sustained release of nerve growth factor from highly homogenous cubosomes stabilized by β-casein with enhanced bioactivity and bioavailability." RSC Advances 6, no. 115 (2016): 114676–84. http://dx.doi.org/10.1039/c6ra19887b.
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Drummond, Heather A., Marise M. Furtado, Samuel Myers, Samira Grifoni, Kimberly A. Parker, Angela Hoover, and David E. Stec. "ENaC proteins are required for NGF-induced neurite growth." American Journal of Physiology-Cell Physiology 290, no. 2 (February 2006): C404—C410. http://dx.doi.org/10.1152/ajpcell.00210.2005.
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Neurite growth is required for nervous system development and repair. Multiple signals, including neurotrophic factors and intact mechanosensing mechanisms, interact to regulate neurite growth. Degenerin/epithelial Na+ channel (DEG/ENaC) proteins have been identified as putative mechanosensors in sensory neurons. Recently, others have shown that the neurotrophic factor NGF stimulates expression of acid-sensing ion channel molecules, which are members of the DEG/ENaC family. However, it is unknown whether NGF regulates ENaC expression or whether ENaC expression is required for neurite formation. Therefore, the aims of the present study were to determine whether ENaC expression is 1) regulated by NGF and 2) required for NGF-induced neurite growth in pheochromocytoma PC-12 cells. We found NGF-induced expression of β- and γ-subunits of ENaC, but not α-ENaC. Tyrosine kinase A (TrkA) receptor blockade abolished NGF-induced β- and γ-ENaC expression and neurite formation. NGF-induced neurite formation was inhibited by disruption of ENaC expression using 1) pharmacological blockade with benzamil, a specific ENaC inhibitor; 2) small interfering RNA; and 3) dominant-negative ENaC molecules. These data indicate NGF-TrkA regulation of ENaC expression may be required for neurite growth and may suggest a novel role for DEG/ENaC proteins in neuronal remodeling and differentiation.
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Sanchez-Rodriguez, A., M. Arias-Alvarez, P. G. Rebollar, P. L. Lorenzo, and R. M. Garcia-Garcia. "148 Inmunolocalization of β-Nerve Growth Factor (NGF) in Male Reproductive Tract and NGF Levels in Serum and Seminal Plasma at Puberty and Adulthood in Rabbit." Reproduction, Fertility and Development 30, no. 1 (2018): 214. http://dx.doi.org/10.1071/rdv30n1ab148.
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β-Nerve growth factor (β-NGF) is a neurotrophin with different roles in reproduction that could be regulated by sexual hormone concentrations. The objective of the present study was to characterise β-NGF in the accessory glands and epididymis of male rabbit and to determine whether its expression differentially changed near puberty (week 22) and adulthood (week 37) according to serum testosterone levels and NGF levels in both serum and seminal plasma (SP). Semen and blood were collected every week from 6 males during 3 months and analysed at 3 time points (weeks 22, 30, and 37) by ELISA. The SP was separated from sperm cells by centrifugation (3,000 × g for 15 min at 4°C) and stored at –20°C; blood was collected in EDTA tubes, centrifuged 15 min at 700 × g at 4°C and stored at –20°C too. Reproductive tissues (prostate, bulbouretral gland, and caput and cauda of epididymis) were collected at the beginning (week 22; n = 4) and at the end (week 37; n = 4) of the experiment. For tissue recovery, males were killed and glands and epididymis were dissected, fixed in modified Bouin’s fluid, and mounted in paraffin. The ELISA for β-NGF (abx259154, Rabbit NGF ELISA kit, Abbexa, Cambridge, UK) and testosterone (DE1559, Demeditec Diagnostics, Kiel, Germany) were performed following the kit protocols. For immunohistochemistry (IHC), goat polyclonal anti-NGF antibody was used in a dilution 1:100 (N8773, Sigma Aldrich, St. Louis, MO, USA). The avidin-biotin-peroxidase complex (ABC) method was performed with the Vectastain ABC kit (Vector Laboratories, Burlingame, CA, USA) and then slides were contrasted with hematoxylin. Results of serum testosterone levels revealed a significant decrease (P < 0.05) at week 37 (0.57 ± 0.18 ng mL−1) from week 30 (4.68 ± 0.86 ng mL−1) and week 22 (3.38 ± 0.99 ng mL−1). However, β-NGF levels at week 37 in serum (299.32 ± 60.22 pg mL−1) and in SP (947.29 ± 249.45 pg mL−1) also decreased but there were no significant differences. Also, we found no correlation between β-NGF in serum and SP and testosterone levels. β-Nerve growth factor was immunolocated in prostate of both ages where epithelial cells were highly stained. At 22 weeks old, stain was located mostly in the apical zone of the cytoplasm, whereas at 37 weeks old, the protein was localised in the entire cytoplasm of the cell. Furthermore, the interstitial tissue in adult males had a moderate stain in contrast with younger males, which did not show signal in that tissue. The content in the prostate lumen was stained too. Bulbourethral glands had a low signal in interstitial tissue that seems to be greater at week 37. Caput and cauda of epididymis of both ages were not stained. These results suggest that β-NGF has different immunolocation in the rabbit male accessory glands depending on the age of animal that could be related to changes in serum testosterone levels. However, these differences were not correlated with β-NGF levels neither in blood serum nor in SP. Research funded by AGL2015-65572-C2-2-R Grant and Predoctoral Contract UCM-Santander.
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Frantsiyants, Elena M., Irina V. Kaplieva, Valeria A. Bandovkina, Ekaterina I. Surikova, Irina V. Neskubina, Lidia K. Trepitaki, Natalia D. Cheryarina, et al. "Neurotrophins in the brain of C57BL/6-Plautm1.1BugThisPlauGFDhu/GFDhu mice with B16/F10 melanoma growing with comorbid pathology." Journal of Clinical Oncology 39, no. 15_suppl (May 20, 2021): e21557-e21557. http://dx.doi.org/10.1200/jco.2021.39.15_suppl.e21557.
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e21557 Background: 10% of cancer patients have comorbidities accompanied by chronic pain. Neurotrophins and fibrinolytic system are involved in carcinogenesis and pain pathogenesis. The purpose of the study was to measure levels of neurotrophins in white matter of the brain of urokinase-deficient (uPA–) mice with B16/F10 melanoma growing in presence of chronic neuropathic pain (CNP). Methods: The study included female mice С57ВL/6 (normal genome uPA+, n = 40) and C57BL/6-Plautm1.1BugThisPlauGFDhu/GFDhu (urokinase gene-knockout uPA–, n = 28) with B16/F10 melanoma (M) implanted subcutaneously 2 weeks after bilateral sciatic nerve ligation (CNP model). Intact mice (I) were controls. Levels of brain-derived neurotrophic factor BDNF, nerve growth factor NGF-β and neurotrophins 3 and 4 (NT3, NT4) were measured by ELISA in white matter of the brain after 3 weeks of tumor growth in presence of CNP. Results: Tumor volume in (uPA–) females by week 3 of carcinogenesis was 0.04 cm3, which was 70 times smaller than in (uPA+) females. Tumor volume in (uPA–) females with CNP was 5.76 cm3, which was 144 times larger than in (uPA–) females without CNP, and in (uPA+) females – 2.5 cm3. The brain of I (uPA–) showed higher levels of NT3 (by 1.3 times, p < 0.05), NT4 (by 2.6 times) and NGF-β (by 1.9 times, p < 0.05) and lower BDNF (by 1.7 times, p < 0.05), compared to I (uPA+). Both strains of mice with M or CNP demonstrated decreased levels of NGF-β, more pronounced in animals with a combination of these factors. (uPA–) females with CNP+M showed a decrease of NT3 and BDNF by 2 times, with NGF-β 2.2 times higher than in (uPA+) mice. Conclusions: The study revealed underlying differences in levels of neurotrophins in the brain of (uPA–) females which could contribute to the creation of conditions for the inhibition of tumor growth. Changes in the levels of NGF-β in mice with melanoma or CNP were nonspecific. Changes in the BDNF, NT3 and NGF-β balance in the brain of (uPA–) mice may be part of the mechanism of greater stimulation of melanoma growth under the influence of CNP.
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Teng, Jian, Zun-Yi Wang, and Dale E. Bjorling. "Estrogen-induced proliferation of urothelial cells is modulated by nerve growth factor." American Journal of Physiology-Renal Physiology 282, no. 6 (June 1, 2002): F1075—F1083. http://dx.doi.org/10.1152/ajprenal.00215.2001.
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Both nerve growth factor (NGF) and estrogen have been shown to stimulate proliferation of various cell types. Human urothelial cells (HUC) express the α- and β-subtypes of the estrogen receptor (ERαand ERβ) as well as tyrosine kinase A (trkA), the high-affinity receptor for NGF. We investigated interactions between estrogen and NGF relative to cell proliferation using primary cultures of HUC. 17β-estradiol (E2) stimulated NGF synthesis by HUC, and E2 (50 nM), the ERαagonist 16α-iodo-17β-estradiol (10 nM), or the ERβagonist genistein (50 nM) each stimulated HUC proliferation, an effect that was abolished by the estrogen antagonist ICI-182,780 (100 nM). NGF (1–100 ng/ml) stimulated HUC proliferation, and this was abolished by NGF antiserum (0.1 μl/ml) or the trkA antagonist K252a (100 nM). HUC proliferation stimulated by E2 was also abolished by NGF antiserum or K252a. Finally, we observed that treatment of HUC with NGF (50 ng/ml) or E2 (50 nM) stimulated trkA phosphorylation, and this was abolished by K252a (100 nM) or NGF antiserum (0.1 μl/ml). These data indicate that the effects of ER activation on HUC proliferation at least partly involve activation of trkA by NGF.
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Garrido, Maritza P., Iván Hurtado, Manuel Valenzuela-Valderrama, Renato Salvatierra, Andrea Hernández, Margarita Vega, Alberto Selman, Andrew F. G. Quest, and Carmen Romero. "NGF-Enhanced Vasculogenic Properties of Epithelial Ovarian Cancer Cells Is Reduced by Inhibition of the COX-2/PGE2 Signaling Axis." Cancers 11, no. 12 (December 7, 2019): 1970. http://dx.doi.org/10.3390/cancers11121970.
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Epithelial ovarian cancer (EOC) is a lethal gynecological neoplasia characterized by extensive angiogenesis and overexpression of nerve growth factor (NGF). Here, we investigated the mechanism by which NGF increases vascular endothelial growth factor (VEGF) expression and the vasculogenic potential of EOC cells, as well as the contribution of the cyclooxygenase 2/prostaglandin E2 (COX-2/PGE2) signaling axis to these events. EOC biopsies and ovarian cell lines were used to determine COX-2 and PGE2 levels, as well as those of the potentially pro-angiogenic proteins c-MYC (a member of the Myc transcription factors family), survivin, and β-catenin. We observed that COX-2 and survivin protein levels increased during EOC progression. In the EOC cell lines, NGF increased the COX-2 and PGE2 levels. In addition, NGF increased survivin, c-MYC, and VEGF protein levels, as well as the transcriptional activity of c-MYC and β-catenin/T-cell factor/lymphoid enhancer-binding factor (TCF-Lef) in a Tropomyosin receptor kinase A (TRKA)-dependent manner. Also, COX-2 inhibition prevented the NGF-induced increases in these proteins and reduced the angiogenic score of endothelial cells stimulated with conditioned media from EOC cells. In summary, we show here that the pro-angiogenic effect of NGF in EOC depends on the COX-2/PGE2 signaling axis. Thus, inhibition COX-2/PGE2 signaling will likely be beneficial in the treatment of EOC.
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Bergfelt, Don R., Jason L. Blum, Jill R. Ratner, Marcelo H. Ratto, Justin K. O'Brien, and Todd R. Robeck. "Preliminary Evaluation of Seminal Plasma Proteins and Immunoreactivity of Nerve Growth Factor as Indicative of an Ovulation Inducing Factor in Odontocetes." Journal of Zoo Biology 2, no. 1 (December 22, 2019): 21–29. http://dx.doi.org/10.33687/zoobiol.002.01.2608.
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In the seminal plasma of terrestrial mammalian species known as induced (e.g., camels) and spontaneous (e.g., cattle) ovulators, an ovulation-inducing factor (OIF) with a protein structure similar to beta-nerve growth factor (β-NGF) has been identified. Detection of an OIF/NGF in the seminal plasma of cetaceans would have both basic and applied implications in reproductive biology and conservation management programs. A preliminary evaluation was conducted to characterize the distribution and abundance of seminal plasma proteins in aquarium-based belugas and a Pacific white-sided and bottlenose dolphin. Initially, SDS-PAGE was used with 50 µg of total protein for separation; thereafter, Western immunoblot was used with anti-NGF. In addition to odontocete seminal plasma, a purified fraction of llama seminal plasma (100 ng protein) and an extract of mouse brain (20 µg total protein) were included as positive controls for NGF. Within the two belugas, visual inspection of the protein bands indicated similar distribution and intensity. However, among the belugas and Pacific white-sided and bottlenose dolphins there was more diversity than similarity in the distribution and abundance of seminal plasma proteins. While immunoreactivity of NGF was distinctly evident in the llama and mouse positive controls, there was no visual reactivity in any of the odontocete samples. These preliminary results provide novel information indicating more homogeneity within and heterogeneity among seminal plasma proteins of ondentocetes. Although NGF was not immunologically detected, future studies are required to address the apparent limitations of immuno-detection of NGF, especially if the post-translational form of β-NGF is in low abundance in the seminal plasma of belugas and Pacific white-sided and bottlenose dolphins.
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Kershaw-Young, C. M., X. Druart, J. Vaughan, and W. M. C. Maxwell. "β-Nerve growth factor is a major component of alpaca seminal plasma and induces ovulation in female alpacas." Reproduction, Fertility and Development 24, no. 8 (2012): 1093. http://dx.doi.org/10.1071/rd12039.
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Ovulation in camelids is induced by an unidentified protein in the seminal plasma of the male termed ‘ovulation-inducing factor’. This protein has been reported to be a 14-kDa protein under reducing conditions, which, when purified from seminal plasma, induces ovulation in llamas. The identification of this protein and investigation of its potential to induce ovulation in camelids may aid the development of protocols for the induction of ovulation. In the present study, alpaca seminal plasma proteins were separated using one-dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis and the most abundant protein of 14 kDa was identified as β-nerve growth factor (β-NGF) by liquid chromatography mass spectrometry. Female alpacas (n = 5 per group) were given intramuscular injections of: (1) 1 mL of 0.9% saline; (2) 4 µg buserelin, a gonadotrophin-releasing hormone agonist; (3) 2 mL alpaca seminal plasma; or (4) 1 mg human β-NGF. Ovulation was detected by transrectal ultrasonography 8 days after treatment and confirmed by plasma progesterone concentrations. Ovulation occurred in 0%, 80%, 80% and 80% of animals treated with saline, buserelin, seminal plasma and β-NGF, respectively. Treatment type did not affect the diameter of the corpus luteum, but plasma progesterone concentrations were lower in saline-treated animals than in the other treatment groups owing to the lack of a corpus luteum. The present study is the first to identify the ovulation-inducing factor protein in alpacas. β-NGF successfully induces ovulation in alpacas and this finding may lead to new methods for the induction of ovulation in camelids.
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Tomlinson, Ryan E., Zhi Li, Zhu Li, Liliana Minichiello, Ryan C. Riddle, Arun Venkatesan, and Thomas L. Clemens. "NGF-TrkA signaling in sensory nerves is required for skeletal adaptation to mechanical loads in mice." Proceedings of the National Academy of Sciences 114, no. 18 (April 17, 2017): E3632—E3641. http://dx.doi.org/10.1073/pnas.1701054114.
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Sensory nerves emanating from the dorsal root extensively innervate the surfaces of mammalian bone, a privileged location for the regulation of biomechanical signaling. Here, we show that NGF-TrkA signaling in skeletal sensory nerves is an early response to mechanical loading of bone and is required to achieve maximal load-induced bone formation. First, the elimination of TrkA signaling in mice harboring mutant TrkAF592A alleles was found to greatly attenuate load-induced bone formation induced by axial forelimb compression. Next, both in vivo mechanical loading and in vitro mechanical stretch were shown to induce the profound up-regulation of NGF in osteoblasts within 1 h of loading. Furthermore, inhibition of TrkA signaling following axial forelimb compression was observed to reduce measures of Wnt/β-catenin activity in osteocytes in the loaded bone. Finally, the administration of exogenous NGF to wild-type mice was found to significantly increase load-induced bone formation and Wnt/β-catenin activity in osteocytes. In summary, these findings demonstrate that communication between osteoblasts and sensory nerves through NGF-TrkA signaling is essential for load-induced bone formation in mice.
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Miyamoto, A., K. Okuda, F. J. Schweigert, and D. Schams. "Effects of basic fibroblast growth factor, transforming growth factor-β and nerve growth factor on the secretory function of the bovine corpus luteum in vitro." Journal of Endocrinology 135, no. 1 (October 1992): 103–14. http://dx.doi.org/10.1677/joe.0.1350103.
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ABSTRACT The effects were investigated of basic fibroblast growth factor (bFGF), transforming growth factor-β (TGF-β) and nerve growth factor (NGF) on the release of progesterone and oxytocin from the bovine corpus luteum (CL) at different stages of the oestrous cycle. A microdialysis system (MDS) of CL and a cell culture system with a reduced number of endothelial cells were used. In the MDS of CL from the mid-luteal stage (days 8–12 of the oestrous cycle), infusion with bFGF (0·1, 1, 10 and 100 ng/ml), TGF-β (0·1, 1 and 10 ng/ml) and NGF (0·1, 1, 10 and 100 ng/ml) for 30 min induced significant acute effects on the release of progesterone. Both bFGF and NGF stimulated the release of progesterone during peptide infusion, TGF-β and also bFGF in the period thereafter. This stimulation was dose-dependent during and after the infusion only for bFGF. This response pattern was observed at all luteal stages for the three growth factors, but bFGF was more stimulatory at the early (days 5–7) and mid-luteal stages during and after peptide infusion. The release of oxytocin was stimulated by bFGF in a dose-dependent manner. At the highest dose, bFGF, TGF-β and NGF stimulated the release of oxytocin throughout all three luteal stages. When luteal cells were cultured with growth factors, only TGF-β showed a dose-dependent inhibition of both basal and LH-stimulated progesterone as well as oxytocin release (measured between 48 and 52 h of culture). NGF had an inhibitory effect only on the basal release of oxytocin. bFGF had no effect on the release of either hormone under continuous stimulation in cell culture. The results indicate that bFGF, TGF-β and NGF act directly and acutely on the secretory function of bovine CL in the MDS but also have long-term effects as shown in cell culture. bFGF appears to be an important autocrine/paracrine regulator of CL function, since local expression of its mRNA, peptide synthesis and its mitogenic and non-mitogenic actions have now been confirmed. Endothelial cells from the CL have been identified as target cells for bFGF. Differences observed between the two systems might thus be attributed to the presence or absence of cellto-cell contact and a reduced number of endothelial cells, as well as to the duration of peptide stimulation and medium changes every 24 h compared with the flow-through conditions in the MDS. Journal of Endocrinology (1992) 135, 103–114
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Ellrich, J., and A. Makowska. "Nerve Growth Factor and ATP Excite Different Neck Muscle Nociceptors in Anaesthetized Mice." Cephalalgia 27, no. 11 (November 2007): 1226–35. http://dx.doi.org/10.1111/j.1468-2982.2007.01431.x.
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Neck muscle nociception probably plays a major role in the pathophysiology of tension-type headache. Recent studies have demonstrated sustained facilitation of brainstem nociception due to noxious neck muscle input evoked by nerve growth factor (NGF) or α,β-methylene ATP (ATP) in mice. Hypothesized different afferent pathways in NGF and ATP models were addressed by local application of tetrodotoxin (TTX) in neck muscles. Brainstem nociception was monitored in 55 anaesthetized mice by the jaw-opening reflex elicited by electrical tongue stimulation. Sole administration of 100nmol/l ATP or 0.8 μmol/l NGF evoked sustained reflex facilitation for at least 95 min. Preceding TTX administration prevented ATP-induced facilitation, but was without effect on NGF. Subsequent administration of 100 nmol/l TTX reversed ATP-evoked facilitation, but was ineffective on NGF. Divergent effects of TTX suggest preferential excitation of group III muscle afferents by ATP and group IV by NGF. Thus, both models address different pathways in pericranial pain.
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Gonçalves, Lucas, Muller C. Martins, Natalia Arle, Rafaela T. Torres, Luisa Migilo, and Marcelo Beletti. "PSX-A-23 Late-Breaking: Supplementation of Nerve Growth Factor (β-NGF) in the maturation medium and efficiency of in vitro embryo production in cattle." Journal of Animal Science 99, Supplement_3 (October 8, 2021): 365. http://dx.doi.org/10.1093/jas/skab235.669.
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Abstract The aim of this study was to evaluate the supplementation of Nerve Growth Factor (β-NGF) in the maturation medium in in vitro embryo production routines. Antral follicles were aspirated from ovaries of cows obtained from slaughterhouses and then oocytes were selected for quality (grades I and II) for in vitro maturation and subjected to 4 successive in vitro embryo production routines (IVEP). Supplementation of 100 ng of β-NGF was performed in the oocyte maturation medium 22 hours before in vitro fertilization. 48 hours after fertilization of the oocytes, an analysis was made of their cleavage rate by counting blastomeres with the aid of a stereoscopic microscope (cleavage rate = number of embryos / number of initial oocytes). Seven days after fertilization, the blastocyst rate was determined through the relation to the number of oocytes that started cleavage and reached this stage of development (blastocyst rate = number of blastocyst / number of oocytes that started cleavage). To verify the existence of a difference between the supplemented and the non-supplemented groups, the paired T test was applied, using the Excel / Action software (Microsoft). In vitro embryo production routines supplemented with β-NGF in the maturation medium had, on average, a higher cleavage rate (P = 0.0072) and a higher blastocyst rate (P = 0.0033) compared to non-supplemented routines with β-NGF. In this study was demonstrated that Nerve Growth Factor supplementation in the maturation medium improves the efficiency of in vitro embryo production in cattle, and this protein has a probable action in the oocyte maturation process.
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Van Kanegan, Michael J., and Stefan Strack. "The Protein Phosphatase 2A Regulatory Subunits B′β and B′δ Mediate Sustained TrkA Neurotrophin Receptor Autophosphorylation and Neuronal Differentiation." Molecular and Cellular Biology 29, no. 3 (November 24, 2008): 662–74. http://dx.doi.org/10.1128/mcb.01242-08.
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ABSTRACT Nerve growth factor (NGF) is critical for the differentiation and maintenance of neurons in the peripheral and central nervous system. Sustained autophosphorylation of the TrkA receptor tyrosine kinase and long-lasting activation of downstream kinase cascades are hallmarks of NGF signaling, yet our knowledge of the molecular mechanisms underlying prolonged TrkA activity is incomplete. Protein phosphatase 2A (PP2A) is a heterotrimeric Ser/Thr phosphatase composed of a scaffolding, catalytic, and regulatory subunit (B, B′, and B" gene families). Here, we employ a combination of pharmacological inhibitors, regulatory subunit overexpression, PP2A scaffold subunit exchange, and RNA interference to show that PP2A containing B′ family regulatory subunits participates in sustained NGF signaling in PC12 cells. Specifically, two neuron-enriched regulatory subunits, B′β and B′δ, recruit PP2A into a complex with TrkA to dephosphorylate the NGF receptor on Ser/Thr residues and to potentiate its intrinsic Tyr kinase activity. Acting at the receptor level, PP2A/ B′β and B′δ enhance NGF (but not epidermal growth factor or fibroblast growth factor) signaling through the Akt and Ras-mitogen-activated protein kinase cascades and promote neuritogenesis and differentiation of PC12 cells. Thus, select PP2A heterotrimers oppose desensitization of the TrkA receptor tyrosine kinase, perhaps through dephosphorylation of inhibitory Ser/Thr phosphorylation sites on the receptor itself, to maintain neurotrophin-mediated developmental and survival signaling.
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Rogdakis, Thanasis, Despoina Charou, Alessia Latorrata, Eleni Papadimitriou, Alexandros Tsengenes, Christina Athanasiou, Marianna Papadopoulou, et al. "Development and Biological Characterization of a Novel Selective TrkA Agonist with Neuroprotective Properties against Amyloid Toxicity." Biomedicines 10, no. 3 (March 6, 2022): 614. http://dx.doi.org/10.3390/biomedicines10030614.
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Neurotrophins are growth factors that exert important neuroprotective effects by preventing neuronal death and synaptic loss. Nerve Growth Factor (NGF) acts through the activation of its high-affinity, pro-survival TrkA and low-affinity, pro-apoptotic p75NTR receptors. NGF has been shown to slow or prevent neurodegenerative signals in Alzheimer’s Disease (AD) progression. However, its low bioavailability and its blood–brain-barrier impermeability limit the use of NGF as a potential therapeutic agent against AD. Based on our previous findings on synthetic dehydroepiandrosterone derivatives, we identified a novel NGF mimetic, named ENT-A013, which selectively activates TrkA and exerts neuroprotective, anti-amyloid-β actions. We now report the chemical synthesis, in silico modelling, metabolic stability, CYP-mediated reaction phenotyping and biological characterization of ENT-A013 under physiological and neurodegenerative conditions. We show that ENT-A013 selectively activates the TrkA receptor and its downstream kinases Akt and Erk1/2 in PC12 cells, protecting these cells from serum deprivation-induced cell death. Moreover, ENT-A013 promotes survival of primary Dorsal Root Ganglion (DRG) neurons upon NGF withdrawal and protects hippocampal neurons against Amyloid β-induced apoptosis and synaptic loss. Furthermore, this neurotrophin mimetic partially restores LTP impairment. In conclusion, ENT-A013 represents a promising new lead molecule for developing therapeutics against neurodegenerative disorders, such as Alzheimer’s Disease, selectively targeting TrkA-mediated pro-survival signals.
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Jung, Eun Joo, Anjugam Paramanantham, Hye Jung Kim, Sung Chul Shin, Gon Sup Kim, Jin-Myung Jung, Soon Chan Hong, Ky Hyun Chung, Choong Won Kim, and Won Sup Lee. "Identification of Growth Factors, Cytokines and Mediators Regulated by Artemisia annua L. Polyphenols (pKAL) in HCT116 Colorectal Cancer Cells: TGF-β1 and NGF-β Attenuate pKAL-Induced Anticancer Effects via NF-κB p65 Upregulation." International Journal of Molecular Sciences 23, no. 3 (January 29, 2022): 1598. http://dx.doi.org/10.3390/ijms23031598.
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The anticancer effects of natural phytochemicals are relevant to the modulation of cytokine signaling pathways in various cancer cells with stem-like properties as well as immune cells. The aim of this study was to elucidate a novel anticancer mechanism of Artemisia annua L. polyphenols (pKAL) involved in the regulation of growth factors, cytokines and mediators in stem-like HCT116 colorectal cancer cells. Through RayBiotech human L-1000 antibody array and bioinformatics analysis, we show here that pKAL-induced anticancer effects are associated with downregulation of growth factor and cytokine signaling proteins including TGFA, FGF16, PDGFC, CCL28, CXCR3, IRF6 and SMAD1. Notably, we found that TGF-β signaling proteins such as GDF10, ENG and TGFBR2 and well-known survival proteins such as NGF-β, VEGFD and insulin were significantly upregulated by pKAL. Moreover, the results of hematoxylin staining, cell viability assay and Western blot analysis demonstrated that TGF-β1 and NGF-β attenuated pKAL-induced anticancer effects by inhibiting pKAL-induced downregulation of caspase-8, NF-κB p65 and cyclin D1. These results suggest that certain survival mediators may be activated by pKAL through the TGF-β1 and NGF-β signaling pathways during pKAL-induced cell death and thus, strategies to inhibit the survival signaling are inevitably required for more effective anticancer effects of pKAL.
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Liu, Jiahao, Jialiu David Li, Jian Lu, Jihong Xing, and Jianhua Li. "Contribution of nerve growth factor to upregulation of P2X3 expression in DRG neurons of rats with femoral artery occlusion." American Journal of Physiology-Heart and Circulatory Physiology 301, no. 3 (September 2011): H1070—H1079. http://dx.doi.org/10.1152/ajpheart.00188.2011.
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Femoral artery occlusion augments the sympathetic nerve and pressor responses to muscle contraction and muscle metabolites injected into the arterial blood supply of the hindlimb muscles in rats. The underlying mechanism by which these reflex responses are enhanced after muscle vascular insufficiency is unclear. Purinergic P2X3 receptor has been reported to contribute to the metabolic component of the exercise pressor reflex. Thus the purpose of this study was to examine if chronic femoral occlusion would alter the expression of P2X3 in dorsal root ganglion (DRG) neurons of rats. Also, P2X3-mediated sympathetic responsiveness was examined after femoral occlusion. In addition, the role played by nerve growth factor (NGF) in regulating the expression and response of P2X3 was examined. Western blot analysis showed that 24 h of femoral ligation increased the levels of P2X3 (optical density: 0.93 ± 0.07 in control and 1.37 ± 0.10 after occlusion; P < 0.05 vs. control). The fluorescence immunohistochemistry further demonstrated that the occlusion elevated P2X3 expression in DRG neurons (percentage of P2X3-positive cells: 33 ± 3% in control and 51 ± 3% in occlusion; P < 0.05 vs. control). Furthermore, the results showed that responses of renal sympathetic nerve activity and blood pressure to stimulation of P2X were greater in occluded rats than responses in control rats by injection of α,β-methylene ATP into the arterial blood supply of the hindlimb muscle. Finally, infusion of NGF in the hindlimb muscles of healthy rats increased P2X3 (optical density: 0.98 ± 0.12 in control and 1.37 ± 0.16 with NGF; P < 0.05 vs. control). The pressor response to injection of α,β-methylene ATP was increased in the rats with NGF infusion. Likewise, blocking NGF attenuated exaggeration of the reflex response induced by α,β-methylene ATP in occluded rats. The findings of this study suggest that the levels of P2X3 in primary afferent neurons are upregulated as the blood supply to the hindlimb is deficient under ischemic conditions, leading to augmentation of the muscle reflex. NGF is closely related to increases in P2X3 receptor expression and response.
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Garrido, Maritza P., Renato Salvatierra, Manuel Valenzuela-Valderrama, Christopher Vallejos, Nicole Bruneau, Andrea Hernández, Margarita Vega, Alberto Selman, Andrew F. G. Quest, and Carmen Romero. "Metformin Reduces NGF-Induced Tumour Promoter Effects in Epithelial Ovarian Cancer Cells." Pharmaceuticals 13, no. 10 (October 16, 2020): 315. http://dx.doi.org/10.3390/ph13100315.
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Epithelial ovarian cancer (EOC) is a lethal gynaecological neoplasm characterized by rapid growth and angiogenesis. Nerve growth factor (NGF) and its high affinity receptor tropomyosin receptor kinase A (TRKA) contribute to EOC progression by increasing the expression of c-MYC, survivin and vascular endothelial growth factor (VEGF) along with a decrease in microRNAs (miR) 23b and 145. We previously reported that metformin prevents NGF-induced proliferation and angiogenic potential of EOC cells. In this study, we sought to obtain a better understanding of the mechanism(s) by which metformin blocks these NGF-induced effects in EOC cells. Human ovarian surface epithelial (HOSE) and EOC (A2780/SKOV3) cells were stimulated with NGF and/or metformin to assess the expression of c-MYC, β-catenin, survivin and VEGF and the abundance of the tumor suppressor miRs 23b and 145. Metformin decreased the NGF-induced transcriptional activity of MYC and β-catenin/T-cell factor/lymphoid enhancer-binding factor (TCF-Lef), as well as the expression of c-MYC, survivin and VEGF in EOC cells, while it increased miR-23b and miR-145 levels. The preliminary analysis of ovarian biopsies from women users or non-users of metformin was consistent with these in vitro results. Our observations shed light on the mechanisms by which metformin may suppress tumour growth in EOC and suggest that metformin should be considered as a possible complementary therapy in EOC treatment.
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Sisman, Ali Rıza, Muge Kiray, Ulas Mehmet Camsari, Merve Evren, Mehmet Ates, Basak Baykara, Ilkay Aksu, Guven Guvendi, and Nazan Uysal. "Potential Novel Biomarkers for Diabetic Testicular Damage in Streptozotocin-Induced Diabetic Rats: Nerve Growth Factor Beta and Vascular Endothelial Growth Factor." Disease Markers 2014 (2014): 1–7. http://dx.doi.org/10.1155/2014/108106.
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Background. It is well known that diabetes mellitus may cause testicular damage. Vascular endothelial growth factor (VEGF) and nerve growth factor beta (NGF-β) are important neurotrophic factors for male reproductive system.Objective. We aimed to investigate the correlation between testicular damage and testicular VEGF and NGF-βlevels in diabetic rats.Methods. Diabetes was induced by streptozotocin (STZ, 45 mg/kg/i.p.) in adult rats. Five weeks later testicular tissue was removed; testicular VEGF and NGF-βlevels were measured by ELISA. Testicular damage was detected by using hematoxylin and eosin staining and periodic acid-Schiff staining, and apoptosis was identified by terminal-deoxynucleotidyl-transferase-mediated dUTP nick end labeling (TUNEL). Seminiferous tubular sperm formation was evaluated using Johnsen’s score.Results. In diabetic rats, seminiferous tubule diameter was found to be decreased; basement membrane was found to be thickened in seminiferous tubules and degenerated germ cells. Additionally, TUNEL-positive cells were increased in number of VEGF+ cells and levels of VEGF and NGF-βwere decreased in diabetic testes. Correlation between VEGF and NGF-βlevels was strong.Conclusion. These results suggest that the decrease of VEGF and NGF-βlevels is associated with the increase of the apoptosis and testicular damage in diabetic rats. Testis VEGF and NGF-βlevels could be potential novel biomarkers for diabetes induced testicular damage.
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Gezginci-Oktayoglu, Selda, and Sehnaz Bolkent. "4-Methlycatechol prevents NGF/p75NTR-mediated apoptosis via NGF/TrkA system in pancreatic β cells." Neuropeptides 45, no. 2 (April 2011): 143–50. http://dx.doi.org/10.1016/j.npep.2011.01.001.
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van Dongen, Robin M., Ronald Zielman, Marek Noga, Olaf M. Dekkers, Thomas Hankemeier, Arn MJM van den Maagdenberg, Gisela M. Terwindt, and Michel D. Ferrari. "Migraine biomarkers in cerebrospinal fluid: A systematic review and meta-analysis." Cephalalgia 37, no. 1 (September 30, 2016): 49–63. http://dx.doi.org/10.1177/0333102415625614.
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Objective To perform a meta-analysis of migraine biomarkers in cerebrospinal fluid (CSF) and of corresponding blood concentrations. Methods We conducted a systematic search for studies that measured biochemical compounds in CSF of chronic or episodic migraineurs and non-headache controls. Subsequent searches retrieved studies with blood measurements of selected CSF biomarkers. If a compound was assessed in three or more studies, results were pooled in a meta-analysis with standardised mean differences (SMD) as effect measures. Results Sixty-two compounds were measured in 40 CSF studies. Most important results include: increased glutamate (five studies, SMD 2.22, 95% CI: 1.30, 3.13), calcitonin gene-related peptide (CGRP) (three studies, SMD: 3.80, 95% CI: 3.19, 4.41) and nerve growth factor (NGF) (three studies, SMD: 6.47, 95% CI: 5.55, 7.39) in chronic migraine patients and decreased β-endorphin (β-EP) in both chronic (four studies, SMD: –1.37, 95% CI: –1.80, –0.94) and interictal episodic migraine patients (three studies, SMD: –1.12, 95% CI: –1.65, –0.58). In blood, glutamate (interictal) and CGRP (chronic, interictal and ictal) were increased and β-EP (chronic, interictal and ictal) was decreased. Conclusions Glutamate, β-EP, CGRP and NGF concentrations are altered in CSF and, except for NGF, also in blood of migraineurs. Future research should focus on the pathophysiological roles of these compounds in migraine.
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Berland, Marco A., Cesar Ulloa-Leal, Miguel Barría, Hollis Wright, Gregory A. Dissen, Mauricio E. Silva, Sergio R. Ojeda, and Marcelo H. Ratto. "Seminal Plasma Induces Ovulation in Llamas in the Absence of a Copulatory Stimulus: Role of Nerve Growth Factor as an Ovulation-Inducing Factor." Endocrinology 157, no. 8 (June 29, 2016): 3224–32. http://dx.doi.org/10.1210/en.2016-1310.
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Llamas are considered to be reflex ovulators. However, semen from these animals is reported to be rich in ovulation-inducing factor(s), one of which has been identified as nerve growth factor (NGF). These findings suggest that ovulation in llamas may be elicited by chemical signals contained in semen instead of being mediated by neural signals. The present study examines this notion. Llamas displaying a preovulatory follicle were assigned to four groups: group 1 received an intrauterine infusion (IUI) of PBS; group 2 received an IUI of seminal plasma; group 3 was mated to a male whose urethra had been surgically diverted (urethrostomized male); and group 4 was mated to an intact male. Ovulation (detected by ultrasonography) occurred only in llamas mated to an intact male or given an IUI of seminal plasma and was preceded by a surge in plasma LH levels initiated within an hour after coitus or IUI. In both ovulatory groups, circulating β-NGF levels increased within 15 minutes after treatment, reaching values that were greater and more sustained in llamas mated with an intact male. These results demonstrate that llamas can be induced to ovulate by seminal plasma in the absence of copulation and that copulation alone cannot elicit ovulation in the absence of seminal plasma. In addition, our results implicate β-NGF as an important mediator of seminal plasma-induced ovulation in llamas because ovulation does not occur if β-NGF levels do not increase in the bloodstream, a change that occurs promptly after copulation with an intact male or IUI of seminal plasma.
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Stewart, J. L., V. R. G. Mercadante, I. F. Canisso, and F. S. Lima. "13 EFFECTS OF NERVE GROWTH FACTOR-β, PURIFIED FROM BULL SEMINAL PLASMA, ON CORPUS LUTEUM FUNCTION AND CONCEPTUS DEVELOPMENT IN COWS." Reproduction, Fertility and Development 29, no. 1 (2017): 114. http://dx.doi.org/10.1071/rdv29n1ab13.
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Nerve growth factor-β (NGF) has been identified in the seminal plasma of livestock and is required for the induction of ovulation in camelids. Until recently, it was thought to play a negligible role in species with spontaneous ovulation. On the contrary, recent studies have shown that systemic administration of NGF, purified from llama seminal plasma, was associated with larger corpus luteum (CL) diameter and higher concentrations of progesterone (P) following ovulation. The objective of the current project was to determine if systemic administration of NGF, purified from bovine seminal plasma, would improve CL formation and enhance embryonic development. Our hypothesis was that systemic administration of NGF at the time of artificial insemination in cows would lead to increased CL volume, increased P secretion, and improved expression of markers of conceptus development and maternal recognition of pregnancy. Seminal plasma was harvested from semen collected by electroejaculation in Angus cross bulls. Purification of NGF was performed using a combination of anion- and cation-exchange chromatography and gradient elution. Beef cows were randomly assigned to CONT (n = 30) or NGF (n = 30) groups and synchronized using a 7-day Co-Synch + CIDR program. At time of insemination (day 0), NGF cows received 296 µg of purified NGF, reconstituted in 12 mL of PBS, and CONT cows received 12 mL of PBS intramuscularly. Blood samples were collected from the coccygeal vein of each cow at days 0, 3, 7, 10, 14, 19, 21, 28, 31, 38, 45, and 66. Ultrasound was performed at each time point for determination of ovarian structures (day 0), corpus luteum volume (all time points), and fetus detection (day 28). Statistical analysis was performed using analysis of variance with repeated-measures in R (R version 3.2.2; https://www.r-project.org/). At day 28, 17/30 (57%) CONT cows and 21/30 (70%) NGF cows were diagnosed as pregnant (P = 0.15). At day 0, cows that later became pregnant had a larger follicular diameter (1.46 cm) than those diagnosed as open (1.16 cm; P < 0.01). Follicular diameter at day 0 did not differ significantly between CONT and NGF groups for either open (P = 0.35) or pregnant (P = 0.90) cows. CL volume in open cows was affected by day (P < 0.001) with no treatment (P = 0.84) or treatment by day (P = 0.42) interaction. CL volume in pregnant cows was affected by both day (P < 0.001) and treatment (P < 0.001), with CONT cows having a higher CL volume than NGF cows. Though NGF appeared to have an inhibitory effect on CL volume in pregnant cows, results are still pending for quantification of P, insulin-like growth factor 1, pregnancy-specific protein B, and interferon-stimulated genes, which will more accurately assess the effects that NGF may have on conceptus development. The results of this study will help us to better understand the role of the seminal plasma protein, NGF, at ovulation and determine if it can be utilised to enhance insemination programs in cattle.
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Ford, Christopher P., Kenneth V. Wong, Van B. Lu, Elena Posse de Chaves, and Peter A. Smith. "Differential Neurotrophic Regulation of Sodium and Calcium Channels in an Adult Sympathetic Neuron." Journal of Neurophysiology 99, no. 3 (March 2008): 1319–32. http://dx.doi.org/10.1152/jn.00966.2007.
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Adult neuronal phenotype is maintained, at least in part, by the sensitivity of individual neurons to a specific selection of neurotrophic factors and the availability of such factors in the neurons' environment. Nerve growth factor (NGF) increases the functional expression of Na+ channel currents ( INa) and both N- and L-type Ca2+ currents ( ICa,N and ICa,L) in adult bullfrog sympathetic ganglion (BFSG) B-neurons. The effects of NGF on ICa involve the mitogen-activated protein kinase (MAPK) pathway. Prolonged exposure to the ganglionic neurotransmitter luteinizing hormone releasing hormone (LHRH) also increases ICa,N but the transduction mechanism remains to be elucidated as does the transduction mechanism for NGF regulation of Na+ channels. We therefore exposed cultured BFSG B-neurons to chicken II LHRH (0.45 μM; 6–9 days) or to NGF (200 ng/ml; 9–10 days) and used whole cell recording, immunoblot analysis, and ras or rap-1 pulldown assays to study effects of various inhibitors and activators of transduction pathways. We found that 1) LHRH signals via ras-MAPK to increase ICa,N, 2) this effect is mediated via protein kinase C-β (PKC-β-ΙΙ), 3) protein kinase A (PKA) is necessary but not sufficient to effect transduction, 4) NGF signals via phosphatidylinositol 3-kinase (PI3K) to increase INa, and 5) long-term exposure to LHRH fails to affect INa. Thus downstream signaling from LHRH has access to the ras-MAPK pathway but not to the PI3K pathway. This allows for differential retrograde and anterograde neurotrophic regulation of sodium and calcium channels in an adult sympathetic neuron.
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Silani, V., A. Pizzuti, A. Falini, G. Borsani, E. I. Rugarli, C. A. Melo, A. Sidoli, F. Villani, F. Baralle, and G. Scarlato. "β-Nerve growth factor (β-NGF) mRNA expression in the Parkinsonian adrenal gland." Experimental Neurology 113, no. 2 (August 1991): 166–70. http://dx.doi.org/10.1016/0014-4886(91)90172-9.
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Mitra, Sumonto, Ruchi Gera, Julia Sundheimer, Marine Lemee, Lars U. Wahlberg, Bengt Linderoth, Maria Eriksdotter, and Homira Behbahani. "Microglia Impairs Proliferation and Induces Senescence In-Vitro in NGF Releasing Cells Used in Encapsulated Cell Biodelivery for Alzheimer’s Disease Therapy." International Journal of Molecular Sciences 23, no. 16 (August 12, 2022): 9011. http://dx.doi.org/10.3390/ijms23169011.
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There is no cure yet available for Alzheimer’s disease (AD). We recently optimized encapsulated cell biodelivery (ECB) devices releasing human mature nerve growth factor (hmNGF), termed ECB-NGF, to the basal forebrain of AD patients. The ECB-NGF delivery resulted in increased CSF cholinergic markers, improved glucose metabolism, and positive effects on cognition in AD patients. However, some ECB-NGF implants showed altered hmNGF release post-explantation. To optimize the ECB-NGF platform for future therapeutic purposes, we initiated in-vitro optimization studies by exposing ECB-NGF devices to physiological factors present within the AD brain. We report here that microglia cells can impair hmNGF release from ECB-NGF devices in-vitro, which can be reversed by transferring the devices to fresh culture medium. Further, we exposed the hmNGF secreting human ARPE-19 cell line (NGC0211) to microglia (HMC3) conditioned medium (MCM; untreated or treated with IL-1β/IFNγ/Aβ40/Aβ42), and evaluated biochemical stress markers (ROS, GSH, ΔΨm, and Alamar Blue assay), cell death indicators (Annexin-V/PI), cell proliferation (CFSE retention and Ki67) and senescence markers (SA-β-gal) in NGC0211 cells. MCMs from activated microglia reduced cell proliferation and induced cell senescence in NGC0211 cells, which otherwise resist biochemical alterations and cell death. These data indicate a critical but reversible impact of activated microglia on NGC0211 cells.
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Frantsiyants, E. M., I. V. Kaplieva, V. A. Bandovkina, E. I. Surikova, I. V. Neskubina, N. D. Cheryarina, L. K. Trepitaki, et al. "The level of neurotrophins in the brain of mice with urokinase gene knockout in experimental melanoma and comorbid pathology." Scientific Notes of the Pavlov University 28, no. 3 (October 25, 2021): 9–16. http://dx.doi.org/10.24884/1607-4181-2021-28-3-9-16.
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The objective was to evaluate the levels of neurotrophins in the brain of mice with urokinase (uPA) gene knockout, carriers of B16/F10 melanoma developing in presence of comorbid pathology – chronic neurogenic pain (CNP).Methods and materials. The study included female mice of two strains: С57ВL/6 (n=40) and C57BL/6-PlautmI.IBug-ThisPlau6FDhu/GFDhu (n=28). In the main groups, CNP was created by the bilateral sciatic nerve ligation, with В16/F10 melanoma transplanted under the skin of the back 2 weeks after. The comparison groups included sham operated animals with melanoma transplantation, the control groups – sham operated animals and animals with CNP. Mice were decapitated on day 21 of the tumor growth, and the brain levels of brain neurotrophic factor (BDNF); nerve growth factor (NGF), neurotrophins 3 (NT3) and 4 (NT4) were studied by ELISA.Results. The brain of mice with uPA gene knockout demonstrated higher levels of NT3 (by 1.3 times (p=0.0146)), NT4 (by 2.6 times) and NGF-β (by 1.9 times (p=0.0021)) and lower BDNF (by 1.7 times (p=0.0203)), compared to mice without knockout. Cerebral reduction of NGF-β was a nonspecific brain response to CNP and neoplastic growth in female mice, enhanced in the combination of the pathological factors. Greater stimulation of subcutaneous melanoma growth in female mice with uPA knockout under the influence of CNP combined with a 2-fold decrease in levels of NT3 and BDNF in the brain, along with 2.2 times higher cerebral levels of NGF-β, compared to female mice without knockout.Conclusions. In female mice with uPA gene knockout compared to mice without knockout, we revealed background differences and other dynamics of neurotrophin levels in the brain at melanoma growth both alone and in combination with comorbid pathology – CNP.
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Berdún, Sergio, Jakub Rychter, and Patri Vergara. "Effects of nerve growth factor antagonist K252a on peritoneal mast cell degranulation: implications for rat postoperative ileus." American Journal of Physiology-Gastrointestinal and Liver Physiology 309, no. 10 (November 15, 2015): G801—G806. http://dx.doi.org/10.1152/ajpgi.00152.2015.
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Stabilization of mast cell (MC) degranulation has been proposed to prevent postoperative ileus (POI). Nerve growth factor (NGF) mediates MC degranulation. The aim of the study was to evaluate whether NGF receptor antagonist K252a acts as a MC stabilizer in vitro and in vivo model of POI. Peritoneal mast cells (PMCs) were obtained from Sprague-Dawley rats and were incubated with K252a and exposed to NGF or Compound 48/80 (C48/80). MC degranulation was assessed by β-hexosaminidase assay. POI was induced in rats by intestinal manipulation (IM). Rats were pretreated with K252a (100 μg/kg sc) 20 min prior to POI induction. At 20 min after IM, release of rat mast cell protease 6 (RMCP-6) was evaluated in peritoneal lavage. At 24 h, intestinal transit (IT) and gastric emptying (GE) were evaluated. Ileal inflammation was assessed by myeloperoxidase (MPO) activity, expression of IL-6, NGF, TrkA, RMCP-2 and 6, and MC density within the full-thickness ileum. C48/80 and NGF evoked degranulation of PMCs in a dose-dependent manner. K252a prevented NGF-evoked, but not C48/80-evoked, MC degranulation. IM evoked the release of peritoneal RMCP-6 and subsequently delayed IT and GE. IM increased MPO activity and expression of IL-6. In IM rats, K252a prevented upregulation of IL-6 expression and reduced TrkA. IT, GE, and inflammation were not affected by K252a. K252a inhibited NGF-evoked degranulation of PMCs in vitro. In vivo, K252a decreased IL-6 and PMC degranulation. This may be of relevance for the development of new therapeutic targets for POI.
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Prautsch, Katharina M., Lucas Degrugillier, Dirk J. Schaefer, Raphael Guzman, Daniel F. Kalbermatten, and Srinivas Madduri. "Ex-Vivo Stimulation of Adipose Stem Cells by Growth Factors and Fibrin-Hydrogel Assisted Delivery Strategies for Treating Nerve Gap-Injuries." Bioengineering 7, no. 2 (May 5, 2020): 42. http://dx.doi.org/10.3390/bioengineering7020042.
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Peripheral nerve injuries often result in lifelong disabilities despite advanced surgical interventions, indicating the urgent clinical need for effective therapies. In order to improve the potency of adipose-derived stem cells (ASC) for nerve regeneration, the present study focused primarily on ex-vivo stimulation of ASC by using growth factors, i.e., nerve growth factor (NGF) or vascular endothelial growth factor (VEGF) and secondly on fibrin-hydrogel nerve conduits (FNC) assisted ASC delivery strategies, i.e., intramural vs. intraluminal loading. ASC were stimulated by NGF or VEGF for 3 days and the resulting secretome was subsequently evaluated in an in vitro axonal outgrowth assay. For the animal study, a 10 mm sciatic nerve gap-injury was created in rats and reconstructed using FNC loaded with ASC. Secretome derived from NGF-stimulated ASC promoted significant axonal outgrowth from the DRG-explants in comparison to all other conditions. Thus, NGF-stimulated ASC were further investigated in animals and found to enhance early nerve regeneration as evidenced by the increased number of β-Tubulin III+ axons. Notably, FNC assisted intramural delivery enabled the improvement of ASC’s therapeutic efficacy in comparison to the intraluminal delivery system. Thus, ex-vivo stimulation of ASC by NGF and FNC assisted intramural delivery may offer new options for developing effective therapies.
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Song, Yizhi, Zunshu Du, Xinyue Chen, Wanning Zhang, Guitao Zhang, Hui Li, Lirong Chang, and Yan Wu. "Astrocytic N-Methyl-D-Aspartate Receptors Protect the Hippocampal Neurons Against Amyloid-β142-Induced Synaptotoxicity by Regulating Nerve Growth Factor." Journal of Alzheimer's Disease 85, no. 1 (January 4, 2022): 167–78. http://dx.doi.org/10.3233/jad-210730.
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Background: Soluble oligomeric amyloid-β (Aβ)-induced synaptic dysfunction is an early event in Alzheimer’s disease (AD) pathogenesis. Mounting evidence has suggested N-methyl-D-aspartate receptors (NMDARs) play an important role in Aβ-induced synaptotoxicity. Originally NMDARs were believed to be expressed exclusively in neurons; however, recent two decades studies have demonstrated functional NMDARs present on astrocytes. Neuronal NMDARs are modulators of neurodegeneration, while our previous initial study found that astrocytic NMDARs mediated synaptoprotection and identified nerve growth factor (NGF) secreted by astrocytes, as a likely mediator, but how astrocytic NMDARs protect neurons against Aβ-induced synaptotoxicity through regulating NGF remains unclear. Objective: To achieve further insight into the mechanism of astrocytic NMDARs oppose Aβ-induced synaptotoxicity through regulating NGF. Methods: With the primary hippocampal neuronal and astrocytic co-cultures, astrocytes were pretreated with agonist or antagonist of NMDARs before Aβ142 oligomers application to neuron-astrocyte co-cultures. Western blot, RT-PCR, etc., were used for the related proteins evaluation. Results: Activation of astrocytic NMDARs can significantly mitigate Aβ142-induced loss of PSD-95 and synaptophysin through increasing NGF release. Blockade of astrocytic NMDARs inhibited Aβ-induced compensatory protective NGF increase in protein and mRNA levels through modulating NF-κB of astrocytes. Astrocytic NMDARs activation can enhance Aβ-induced Furin increase, and blockade of astrocytic NMDARs inhibited Aβ-induced immunofluorescent intensity elevation of vesicle trafficking protein VAMP3 and NGF double-staining. Conclusion: Astrocytic NMDARs oppose Aβ-induced synaptotoxicity through modulating the synthesis, maturation, and secretion of NGF in astrocytes. This new information may contribute to the quest for specific targeted strategy of intervention to delay the onset of AD.
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Zhang, Paul J., Randal Weber, Ho-Hi Liang, Teresa L. Pasha, and Virginia A. LiVolsi. "Growth Factors and Receptors in Juvenile Nasopharyngeal Angiofibroma and Nasal Polyps: An Immunohistochemical Study." Archives of Pathology & Laboratory Medicine 127, no. 11 (November 1, 2003): 1480–84. http://dx.doi.org/10.5858/2003-127-1480-gfarij.
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Abstract Background.—Juvenile nasopharyngeal angiofibroma is a rare nasopharyngeal tumor that occurs exclusively in adolescent boys. It is a histologically benign but locally persistent growth of stromal and vascular tissue. Although male hormones and some growth factors, such as transforming growth factor β1 (TGF-β1), insulin-like growth factor II (IGF-II), and, lately, the proto-oncogene β-catenin, have been implicated in the histogenesis of the tumor, the biologic signaling pathways that drive this peculiar fibrovascular proliferation are still nuclear. Objective.—To evaluate immunoexpressions of β-catenin, c-Kit, p130Cas, TGF-β3, bone morphogenic protein 4, nerve growth factor (NGF), and the IGF receptor (IGF-1R) in a series of juvenile nasopharyngeal angiofibromas and to compare to that of a group of nasal polyps. Design.—A standard immunohistochemical technique was used on paraffin sections of 12 sporadic juvenile nasopharyngeal angiofibromas and 15 nasal polyps with microwave or steam antigen retrieval. Immunoreactivity was analyzed semiquantitatively in stromal cells and endothelial cells of each case. Results.—The expressions of β-catenin (nuclear), c-Kit (cytoplasmic), and NGF (cytoplasmic) were higher and more frequent in stromal cells of juvenile nasopharyngeal angiofibromas than those of nasal polyps. Both juvenile nasopharyngeal angiofibromas and nasal polyps showed similarly frequent and strong immunoreactivity for p130Cas and TGF-β3 and weak immunoreactivity for bone morphogenic protein 4 in both stromal cells and endothelial cells. No IGF-1R immunoreactivity was detected in any case of either group. Conclusions.—Our results support the role of β-catenin in juvenile nasopharyngeal angiofibromas and suggest a potential involvement of c-Kit and NGF signaling pathways in the juvenile nasopharyngeal angiofibromas. Although the biologic significance of c-Kit in juvenile nasopharyngeal angiofibromas has yet to be defined, the finding of frequent and high c-Kit expression might have therapeutic importance for patients with juvenile nasopharyngeal angiofibromas.
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Lei, Saobo, William F. Dryden, and Peter A. Smith. "Nerve Growth Factor Regulates Sodium But Not Potassium Channel Currents in Sympathetic B Neurons of Adult Bullfrogs." Journal of Neurophysiology 86, no. 2 (August 1, 2001): 641–50. http://dx.doi.org/10.1152/jn.2001.86.2.641.
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The TTX-sensitive and -resistant components of the voltage-gated Na+ current (TTX-s I Na and TTX-r I Na) are increased within 2 wk of cutting the axons of B-cells in bullfrog paravertebral sympathetic ganglia (BFSG). Axotomy also increases the noninactivating, voltage-activated K+ current (M current I M), whereas delayed rectifier K+ current ( I K) is reduced. We found that similar effects were produced when BFSG B cells were dissociated from adult bullfrogs and maintained in a defined-medium, neuron-enriched, low-density, serum-free culture. Thus the density of TTX-s I Na, TTX-r I Na, and I M were transiently increased, whereas I K density was decreased. Reduction in voltage-sensitive, Ca2+-dependent K+ current ( I C) was attributed to previously documented decreases in Ca2+ channel current ( I Ca). To test whether axotomy- or culture-induced changes in ion channel function reflect loss of retrograde influence of nerve growth factor (NGF), we examined the effect of murine β-NGF on TTX-s I Na, TTX-r I Na, I K, and I M. Culture of neurons for 15 days in the presence of NGF (200 ng/ml), more than doubled total I Na density but did not enhance neurite outgrowth. The TTX-r I Nadensity was increased about threefold and the TTX-s I Na density increased 2.4-fold. NGF did not affect the activation or inactivation kinetics of the total Na+ conductance. Effects of NGF were blocked by the transcription inhibitors, cordycepin (20 μM) and actinomycin D (0.01 μg/ml). I K and I M were unaffected by NGF, and although I C was enhanced, this likely reflected the known effect of NGF on I Ca in BFSG neurons. Na+ channel synthesis and/or expression in adult sympathetic neurons is therefore subject to selective regulation by NGF. Despite this, the increase in I Na and I M as well as the decrease in I K seen in BFSG neurons in culture or after axotomy cannot readily be explained in terms of alterations in the availability of target-derived NGF.
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Amiss, E., J. W. Stewart, V. M. Negrón-Pérez, K. Jones, H. Haines, M. L. Rhoads, F. S. Lima, and J. L. Stewart. "117 Supplementation of IVF medium with nerve growth factor improved bovine embryonic cleavage rates during summer months." Reproduction, Fertility and Development 32, no. 2 (2020): 185. http://dx.doi.org/10.1071/rdv32n2ab117.
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Nerve growth factor-β (NGF), a protein originally associated with regulation of neuron development, has been found to play a role in the reproductive system of mammals. Previous research showed that administration of NGF to cows resulted in enhanced conceptus development. Although these effects were speculated to be a result of improved corpus luteum function, whether NGF could act directly on the embryo remained undetermined. Therefore, the direct effects of NGF on fertilization and embryo development warrant investigation to see whether it can be used as a novel tool to improve cleavage and blastocyst rates when producing embryos via IVF during periods of suboptimal oocyte quality, such as with heat stress. The objective of this study was to explore how supplementation of NGF, purified from bull seminal plasma, during IVF may directly affect embryo development in oocytes harvested in the summer. Abattoir-derived bovine ovaries were used for recovery of cumulus-oocyte complexes (COCs) over eight replicates through May and June. On Day −1, COCs were collected and matured for 20h in oocyte maturation medium incubated at 38.5°C. On Day 0, matured oocytes were added to a solution of IVF-Tyrode's albumin lactate pyruvate (TALP) and either phosphate-buffered saline (PBS; control) or 100ngmL−1 NGF. Pooled frozen-thawed semen from two different bulls per replicate were added to the IVF solutions and incubated with COCs for 20h at 38.5°C in a humidified atmosphere of 5% CO2. On Day 1, zygotes were washed in HEPES-TALP, and cumulus cells were removed using 1% hyaluronidase. The zygotes were plated in synthetic oviductal fluid (SOF-BE2) culture medium and incubated at 38.5°C in a tri-gas chamber (5% CO2, 5% O2, and balanced N2). Cleavage rates were recorded at 24 and 48h, calculated by dividing the number of cleaved embryos by the total zygote count. Embryos were incubated until Day 8, when the rate of blastocysts was assessed. This study found that the treatment of IVF medium with NGF increased the cleavage rate of embryos after 48h (Control: 59%; NGF: 66%; P=0.04) and the hatched blastocyst percentage per oocyte on Day 8 (Control: 6.7%; NGF: 9.4%; P=0.01). The treatment did not affect the percentage of blastocysts per cleaved embryos (Control: 21%; NGF: 22%; P=0.16) or the hatched blastocyst rate at Day 8 (Control: 53%; NGF: 55%; P=0.67). These results show that NGF can act directly on the oocyte during fertilization to alter subsequent development, specifically through increased embryonic cleavage rates. Further studies are needed to assess different dosages of NGF in order to mitigate the detrimental effects of heat stress on oocyte competence for use in IVF. Follow-up studies using a whole-animal model are needed to understand the clinical relevance of these findings in the ability of embryos to promote maternal recognition of pregnancy.
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Lei, Saobo, William F. Dryden, and Peter A. Smith. "Regulation of N- and L-Type Ca2+ Channels in Adult Frog Sympathetic Ganglion B Cells by Nerve Growth Factor In Vitro and In Vivo." Journal of Neurophysiology 78, no. 6 (December 1, 1997): 3359–70. http://dx.doi.org/10.1152/jn.1997.78.6.3359.
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Lei, Saobo, William F. Dryden, and Peter A. Smith. Regulation of N- and L-type Ca2+ channels in adult frog sympathetic ganglion B cells by nerve growth factor in vitro and in vivo. J. Neurophysiol. 78: 3359–3370, 1997. To examine mechanisms responsible for the long-term regulation of Ca2+-channels in an adult neuron, changes in whole cell Ba2+ current ( I Ba) were examined in adult bullfrog sympathetic ganglion B cells in vitro. Cells were cultured at low density in defined, serum free medium. After 15 days, total I Ba was similar to the initial value, whereas I Ba density was reduced by ∼36%, presumably due to an increase in neuronal surface area. By contrast, I Ba density remained constant after 6–15 days in the presence of murine β-NGF (200 ng/ml), and total I Ba was almost doubled. Inclusion of cytosine arabinoside (Ara-C; 10 μM) to inhibit proliferation of nonneuronal cells, did not affect the survival of neurons in the absence of nerve growth factor (NGF) nor did it attenuate I Ba. Ara-C did not prevent the effect of NGF on I Ba. There were three independent components to the action of NGF; during 6–9 days, it increased ω-conotoxin-GVIA–sensitive N-type I Ba ( I Ba,N); increased nifedipine-sensitive L-type I Ba ( I Ba,L) and decreased inactivation of the total Ba2+ conductance ( g Ba). The latter effect involved a selective decrease in the amplitude of one of the four kinetic components that describe the inactivation process. Total I Ba was also 55.8% larger than control in the somata of B cells acutely dissociated from leopard frogs that had received prior subcutaneous injections of NGF. By contrast, injection of NGF antiserum decreased total I Ba by 29.4%. There was less inactivation of g Ba in B cells from NGF-injected animals than in cells from animals injected with NGF antiserum ( P < 0.001). These data suggest that NGF-like molecule(s) play(s) a role in the maintenance of I Ba in an adult amphibian sympathetic neuron; the presence of NGF may allow the neuron to maintain a constant relationship between cell size and current density. They also show that I Ba inactivation in an adult neuron can be modulated in a physiologically relevant way by an extracellular ligand.
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Magaz, Adrián, Mark D. Ashton, Rania M. Hathout, Xu Li, John G. Hardy, and Jonny J. Blaker. "Electroresponsive Silk-Based Biohybrid Composites for Electrochemically Controlled Growth Factor Delivery." Pharmaceutics 12, no. 8 (August 7, 2020): 742. http://dx.doi.org/10.3390/pharmaceutics12080742.
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Stimuli-responsive materials are very attractive candidates for on-demand drug delivery applications. Precise control over therapeutic agents in a local area is particularly enticing to regulate the biological repair process and promote tissue regeneration. Macromolecular therapeutics are difficult to embed for delivery, and achieving controlled release over long-term periods, which is required for tissue repair and regeneration, is challenging. Biohybrid composites incorporating natural biopolymers and electroconductive/active moieties are emerging as functional materials to be used as coatings, implants or scaffolds in regenerative medicine. Here, we report the development of electroresponsive biohybrid composites based on Bombyx mori silkworm fibroin and reduced graphene oxide that are electrostatically loaded with a high-molecular-weight therapeutic (i.e., 26 kDa nerve growth factor-β (NGF-β)). NGF-β-loaded composite films were shown to control the release of the drug over a 10-day period in a pulsatile fashion upon the on/off application of an electrical stimulus. The results shown here pave the way for personalized and biologically responsive scaffolds, coatings and implantable devices to be used in neural tissue engineering applications, and could be translated to other electrically sensitive tissues as well.
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Safieh-Garabedian, B., G. M. Mouneimne, W. El-Jouni, M. Khattar, and R. Talhouk. "The effect of endotoxin on functional parameters of mammary CID-9 cells." Reproduction 127, no. 3 (March 2004): 397–406. http://dx.doi.org/10.1530/rep.1.00135.
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The effect of endotoxin on mammary CID-9 cells, which differentiate in culture and express β-casein, was investigated. Cells in culture supplemented with lactogenic hormones and dripped with EMS-Matrix (EMS-drip), were treated daily with endotoxin (0.5–500 μg/ml). Endotoxin at concentrations of less or equal to 10 μg/ml did not affect cell growth and viability up to 5 days post endotoxin treatment. Endotoxin (0.01–10 μg/ml) was added to the culture medium, upon confluence, and functional parameters were examined within 48 h post endotoxin treatment. Nuclear factor-κB (NF-κB) (p52) increased in nuclear extracts from endotoxin-stimulated cells within 1 h of treatment, while β-casein mRNA and protein expression decreased in a concentration-dependent manner at 24 and 48 h post treatment. Zymography showed that the 72 and 92 kDa gelatinase activity increased in cells at 24 and 48 h post endotoxin treatment at 10 and 50 μg/ml. At the latter concentration, the active form of 72 kDa gelatinase was induced at 48 h. Interleukin-6 and tumor necrosis factor-α levels increased at 1–3 h post endotoxin treatment and peaked at 6 h in cells on plastic and EHS-drip. Nerve growth factor (NGF) levels increased in control and endotoxin-treated cells in a time-dependent manner, and endotoxin increased NGF levels in culture at 6 and 9 h post endotoxin treatment. This study shows that endotoxin activated NF-κB, suppressed β-casein expression and upregulated gelatinases, cytokines and NGF. This model could be used to investigate the role of mammary cells in initiating and propagating inflammation and to test candidate molecules for potential anti-inflammatory properties.
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Lei, Saobo, William F. Dryden, and Peter A. Smith. "Involvement of Ras/MAP Kinase in the Regulation of Ca2+ Channels in Adult Bullfrog Sympathetic Neurons by Nerve Growth Factor." Journal of Neurophysiology 80, no. 3 (September 1, 1998): 1352–61. http://dx.doi.org/10.1152/jn.1998.80.3.1352.
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Lei, Saobo, William F. Dryden, and Peter A. Smith. Involvement of Ras/MAP kinase in the regulation of Ca2+ channels in adult bullfrog sympathetic neurons by nerve growth factor. J. Neurophysiol. 80: 1352–1361, 1998. The cellular mechanisms that underlie nerve growth factor (NGF) induced increase in Ca2+-channel current in adult bullfrog sympathetic B-neurons were examined by whole cell recording techniques. Cells were maintained at low density in neuron-enriched, defined-medium, serum-free tissue culture for 6 days in the presence or absence of NGF (200 ng/ml). The increase in Ba2+ current ( I Ba) density induced by NGF was attenuated by the RNA synthesis inhibitor cordycepin (20 μM), by the DNA transcription inhibitor actinomycin D (0.01 μg/ml), by inhibitors of Ras isoprenylation (perillic acid 0.1–1.0 mM or α-hydroxyfarnesylphosphonic acid 10–100 μM), by tyrosine kinase inhibitors genistein (20 μM) or lavendustin A (1 μM), and by PD98059 (10–100 μM), an inhibitor of mitogen-activated protein kinase kinase. Inhibitors of the phosphatidylinositol 3-kinase (PI3K) pathway (wortmannin, 100 nM, or LY29400, 100 μM) were ineffective as were inhibitors of phospholipase Cγ (U73122 or neomycin, both 100 μM). The effect of NGF persisted in Ca2+-free medium that contained 1.8 mM Mg2+ and 2 mM ethylene glycol-bis(β-aminoethyl ether)- N, N, N′, N′-tetraacetic acid. It was mimicked by a Trk antibody that was capable of inducing neurite outgrowth in explant cultures of bullfrog sympathetic ganglion. Antibodies raised against the low-affinity p75 neurotrophin receptor were ineffective in blocking the effect of NGF on I Ba. These results suggest that NGF-induced increase in Ca2+ channel current in adult sympathetic neurons results, at least in part, from new channel synthesis after Trk activation of Ras and mitogen activated protein kinase by a mechanism that is independent of extracellular Ca2+.
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Okur, Zeynep, Oya I. Senturk, Canelif Yilmaz, Gulcihan Gulseren, Busra Mammadov, Mustafa O. Guler, and Ayse B. Tekinay. "Promotion of neurite outgrowth by rationally designed NGF-β binding peptide nanofibers." Biomaterials Science 6, no. 7 (2018): 1777–90. http://dx.doi.org/10.1039/c8bm00311d.
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Sari, Luciana M., Renato Zampini, Martin E. Argañaraz, María I. Carretero, Fernanda G. Fumuso, Daniela E. Barraza, Marcelo Ratto, and Silvana A. Apichela. "Expression of β‐NGF and high‐affinity NGF receptor (TrKA) in llama ( Lama glama ) male reproductive tract and spermatozoa." Molecular Reproduction and Development 85, no. 12 (November 14, 2018): 934–44. http://dx.doi.org/10.1002/mrd.23075.
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Sun, Wenjie, Changkai Sun, Hui Zhao, Hang Lin, Qianqian Han, Jingyu Wang, Hui Ma, Bing Chen, Zhifeng Xiao, and Jianwu Dai. "Improvement of Sciatic Nerve Regeneration Using Laminin-Binding Human NGF-β." PLoS ONE 4, no. 7 (July 9, 2009): e6180. http://dx.doi.org/10.1371/journal.pone.0006180.
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