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Academic literature on the topic 'TRP receptors'

Author: Grafiati

Published: 10 March 2023

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Journal articles on the topic "TRP receptors"

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Hu, Jinghua, Samuel G. Wittekind, and Maureen M. Barr. "STAM and Hrs Down-Regulate Ciliary TRP Receptors." Molecular Biology of the Cell 18, no. 9 (September 2007): 3277–89. http://dx.doi.org/10.1091/mbc.e07-03-0239.

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Cilia are endowed with membrane receptors, channels, and signaling components whose localization and function must be tightly controlled. In primary cilia of mammalian kidney epithelia and sensory cilia of Caenorhabditis elegans neurons, polycystin-1 (PC1) and transient receptor polycystin-2 channel (TRPP2 or PC2), function together as a mechanosensory receptor-channel complex. Despite the importance of the polycystins in sensory transduction, the mechanisms that regulate polycystin activity and localization, or ciliary membrane receptors in general, remain poorly understood. We demonstrate that signal transduction adaptor molecule STAM-1A interacts with C. elegans LOV-1 (PC1), and that STAM functions with hepatocyte growth factor–regulated tyrosine kinase substrate (Hrs) on early endosomes to direct the LOV-1-PKD-2 complex for lysosomal degradation. In a stam-1 mutant, both LOV-1 and PKD-2 improperly accumulate at the ciliary base. Conversely, overexpression of STAM or Hrs promotes the removal of PKD-2 from cilia, culminating in sensory behavioral defects. These data reveal that the STAM-Hrs complex, which down-regulates ligand-activated growth factor receptors from the cell surface of yeast and mammalian cells, also regulates the localization and signaling of a ciliary PC1 receptor-TRPP2 complex.

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Manolache, Alexandra, Teodora Stratulat, and Alexandru Babeș. "Modulation of Transient Receptor Potential (TRP) channels by tyrosine phosphorylation." Reviews in Biological and Biomedical Sciences 3, no. 1 (July 4, 2020): 77–87. http://dx.doi.org/10.31178/rbbs.2020.3.1.5.

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Transient Receptor Potential (TRP) channels are a superfamily of polymodal, non-selective receptors, expressed in the nervous system and several other tissues, where they play many physiological or pathological roles. TRP channels are sensitive to a diverse range of stimuli, such as temperature, osmolarity, oxidative stress, external compounds and intracellular signaling molecules. The activity of TRP channels can be modulated by protein phosphorylation, including tyrosine phosphorylation. In this review, we present the studies carried out so far regarding the modulation of TRP channels by tyrosine phosphorylation.

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Lezama-García, Karina, Daniel Mota-Rojas, Alfredo M. F. Pereira, Julio Martínez-Burnes, Marcelo Ghezzi, Adriana Domínguez, Jocelyn Gómez, et al. "Transient Receptor Potential (TRP) and Thermoregulation in Animals: Structural Biology and Neurophysiological Aspects." Animals 12, no. 1 (January 2, 2022): 106. http://dx.doi.org/10.3390/ani12010106.

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This review presents and analyzes recent scientific findings on the structure, physiology, and neurotransmission mechanisms of transient receptor potential (TRP) and their function in the thermoregulation of mammals. The aim is to better understand the functionality of these receptors and their role in maintaining the temperature of animals, or those susceptible to thermal stress. The majority of peripheral receptors are TRP cation channels formed from transmembrane proteins that function as transductors through changes in the membrane potential. TRP are classified into seven families and two groups. The data gathered for this review include controversial aspects because we do not fully know the mechanisms that operate the opening and closing of the TRP gates. Deductions, however, suggest the intervention of mechanisms related to G protein-coupled receptors, dephosphorylation, and ligands. Several questions emerge from the review as well. For example, the future uses of these data for controlling thermoregulatory disorders and the invitation to researchers to conduct more extensive studies to broaden our understanding of these mechanisms and achieve substantial advances in controlling fever, hyperthermia, and hypothermia.

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Zaccor, Nicholas W., Charlotte J. Sumner, and Solomon H. Snyder. "The nonselective cation channel TRPV4 inhibits angiotensin II receptors." Journal of Biological Chemistry 295, no. 29 (June 3, 2020): 9986–97. http://dx.doi.org/10.1074/jbc.ra120.014325.

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G-protein–coupled receptors (GPCRs) are a ubiquitously expressed family of receptor proteins that regulate many physiological functions and other proteins. They act through two dissociable signaling pathways: the exchange of GDP to GTP by linked G-proteins and the recruitment of β-arrestins. GPCRs modulate several members of the transient receptor potential (TRP) channel family of nonselective cation channels. How TRP channels reciprocally regulate GPCR signaling is less well-explored. Here, using an array of biochemical approaches, including immunoprecipitation and fluorescence, calcium imaging, phosphate radiolabeling, and a β-arrestin–dependent luciferase assay, we characterize a GPCR–TRP channel pair, angiotensin II receptor type 1 (AT1R), and transient receptor potential vanilloid 4 (TRPV4), in primary murine choroid plexus epithelial cells and immortalized cell lines. We found that AT1R and TRPV4 are binding partners and that activation of AT1R by angiotensin II (ANGII) elicits β-arrestin–dependent inhibition and internalization of TRPV4. Activating TRPV4 with endogenous and synthetic agonists inhibited angiotensin II–mediated G-protein–associated second messenger accumulation, AT1R receptor phosphorylation, and β-arrestin recruitment. We also noted that TRPV4 inhibits AT1R phosphorylation by activating the calcium-activated phosphatase calcineurin in a Ca2+/calmodulin–dependent manner, preventing β-arrestin recruitment and receptor internalization. These findings suggest that when TRP channels and GPCRs are co-expressed in the same tissues, many of these channels can inhibit GPCR desensitization.

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Rigano, Daniela, Carmen Formisano, and Orazio Taglialatela-Scafati. "Marine Metabolites Modulating CB Receptors and TRP Channels." Planta Medica 82, no. 09/10 (March 22, 2016): 761–66. http://dx.doi.org/10.1055/s-0042-101352.

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Maliszewska, Justyna, Milena Jankowska, Hanna Kletkiewicz, Maria Stankiewicz, and Justyna Rogalska. "Effect of Capsaicin and Other Thermo-TRP Agonists on Thermoregulatory Processes in the American Cockroach." Molecules 23, no. 12 (December 18, 2018): 3360. http://dx.doi.org/10.3390/molecules23123360.

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Capsaicin is known to activate heat receptor TRPV1 and induce changes in thermoregulatory processes of mammals. However, the mechanism by which capsaicin induces thermoregulatory responses in invertebrates is unknown. Insect thermoreceptors belong to the TRP receptors family, and are known to be activated not only by temperature, but also by other stimuli. In the following study, we evaluated the effects of different ligands that have been shown to activate (allyl isothiocyanate) or inhibit (camphor) heat receptors, as well as, activate (camphor) or inhibit (menthol and thymol) cold receptors in insects. Moreover, we decided to determine the effect of agonist (capsaicin) and antagonist (capsazepine) of mammalian heat receptor on the American cockroach’s thermoregulatory processes. We observed that capsaicin induced the decrease of the head temperature of immobilized cockroaches. Moreover, the examined ligands induced preference for colder environments, when insects were allowed to choose the ambient temperature. Camphor exposure resulted in a preference for warm environments, but the changes in body temperature were not observed. The results suggest that capsaicin acts on the heat receptor in cockroaches and that TRP receptors are involved in cockroaches’ thermosensation.

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Ojiro, Ichie, Hiromi Nishio, Toyomi Yamazaki-Ito, Shogo Nakano, Sohei Ito, Yoshikazu Toyohara, Tadahiro Hiramoto, Yuko Terada, and Keisuke Ito. "Trp-Trp acts as a multifunctional blocker for human bitter taste receptors, hTAS2R14, hTAS2R16, hTAS2R43, and hTAS2R46." Bioscience, Biotechnology, and Biochemistry 85, no. 6 (April 12, 2021): 1526–29. http://dx.doi.org/10.1093/bbb/zbab061.

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ABSTRACT Many functional food ingredients activate human bitter taste receptors (hTAS2Rs). In this study, A novel inhibitor, Trp-Trp, for hTAS2R14 was identified by searching for the agonist peptide's analogs. Trp-Trp also inhibited hTAS2R16, hTAS2R43, and hTAS2R46, which share the same agonists with hTAS2R14. The multifunctional characteristic of Trp-Trp is advantageous for use as bitterness-masking agents in functional foods.

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Thapa, Dibesh, Brentton Barrett, Fulye Argunhan, and Susan D. Brain. "Influence of Cold-TRP Receptors on Cold-Influenced Behaviour." Pharmaceuticals 15, no. 1 (December 28, 2021): 42. http://dx.doi.org/10.3390/ph15010042.

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The transient receptor potential (TRP) channels, TRPA1 and TRPM8, are thermo-receptors that detect cold and cool temperatures and play pivotal roles in mediating the cold-induced vascular response. In this study, we investigated the role of TRPA1 and TRPM8 in the thermoregulatory behavioural responses to environmental cold exposure by measuring core body temperature and locomotor activity using a telemetry device that was surgically implanted in mice. The core body temperature of mice that were cooled at 4 °C over 3 h was increased and this was accompanied by an increase in UCP-1 and TRPM8 level as detected by Western blot. We then established an effective route, by which the TRP antagonists could be administered orally with palatable food. This avoids the physical restraint of mice, which is crucial as that could influence the behavioural results. Using selective pharmacological antagonists A967079 and AMTB for TRPA1 and TRPM8 receptors, respectively, we show that TRPM8, but not TRPA1, plays a direct role in thermoregulation response to whole body cold exposure in the mouse. Additionally, we provide evidence of increased TRPM8 levels after cold exposure which could be a protective response to increase core body temperature to counter cold.

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Fernandes, E. S., S. Awal, R. Karadaghi, and S. D. Brain. "TRP Receptors in Arthritis, Gaining Knowledge for Translation from Experimental Models." Open Pain Journal 6, no. 1 (March 8, 2013): 50–61. http://dx.doi.org/10.2174/1876386301306010050.

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Arthritis is a condition characterised by mainly pain, reduced joint movement and signs of inflammation, such as swelling. The disorder has many different types, of which osteoarthritis (a degenerative joint disease) and rheumatoid arthritis (a chronic autoimmune disease) are the two most common forms. There are >6 million sufferers in the UK and both conditions have a huge potential to impair capabilities and contribute to social and economic burdens. Whilst there are a wide range of arthritic therapies available, many patients under treatment complain of poor pain relief. Thus there is a need for novel therapeutic approaches, and the transient receptor potential (TRP) family of receptor channels has been investigated. One particular area of recent research has been the ligand-gated transient receptor potential vanilloid 1 (TRPV1) channel. Findings from numerous pre-clinical models and scientific studies have shown that TRPV1 desensitisation, or the use of TRPV1 antagonists alleviates pain and some inflammatory aspects. New findings have started to unveil the potential of other TRP channels in mediating arthritic pain and inflammation. With the understanding that the currently available treatments for arthritis are limited, researchers have looked into the exciting prospect that TRP receptor antagonists may be developed into effective, specific drugs, which would potentially protect against the complications of arthritis. These antagonists are still under development, although only data from studies from pre-clinical models are currently available. This review acts to summarize knowledge of the potential influence of TRP receptors in arthritis to date.

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Shabir, Saqib, William Cross, Lisa A. Kirkwood, Joanna F. Pearson, Peter A. Appleby, Dawn Walker, Ian Eardley, and Jennifer Southgate. "Functional expression of purinergic P2 receptors and transient receptor potential channels by the human urothelium." American Journal of Physiology-Renal Physiology 305, no. 3 (August 1, 2013): F396—F406. http://dx.doi.org/10.1152/ajprenal.00127.2013.

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In addition to its role as a physical barrier, the urothelium is considered to play an active role in mechanosensation. A key mechanism is the release of transient mediators that activate purinergic P2 receptors and transient receptor potential (TRP) channels to effect changes in intracellular Ca2+. Despite the implied importance of these receptors and channels in urothelial tissue homeostasis and dysfunctional bladder disease, little is known about their functional expression by the human urothelium. To evaluate the expression and function of P2X and P2Y receptors and TRP channels, the human ureter and bladder were used to separate urothelial and stromal tissues for RNA isolation and cell culture. RT-PCR using stringently designed primer sets was used to establish which P2 and TRP species were expressed at the transcript level, and selective agonists/antagonists were used to confirm functional expression by monitoring changes in intracellular Ca2+ and in a scratch repair assay. The results confirmed the functional expression of P2Y4 receptors and excluded nonexpressed receptors/channels (P2X1, P2X3, P2X6, P2Y6, P2Y11, TRPV5, and TRPM8), while a dearth of specific agonists confounded the functional validation of expressed P2X2, P2X4, P2Y1, P2Y2, TRPV2, TRPV3, TRPV6 and TRPM7 receptors/channels. Although a conventional response was elicited in control stromal-derived cells, the urothelial cell response to well-characterized TRPV1 and TRPV4 agonists/antagonists revealed unexpected anomalies. In addition, agonists that invoked an increase in intracellular Ca2+ promoted urothelial scratch repair, presumably through the release of ATP. The study raises important questions about the ligand selectivity of receptor/channel targets expressed by the urothelium. These pathways are important in urothelial tissue homeostasis, and this opens the possibility of selective drug targeting.

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Dissertations / Theses on the topic "TRP receptors"

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Jones, Christopher M. "Expression and folding studies of the ankyrin repeat domain of the capsaicin receptor." Click here for download, 2006. http://wwwlib.umi.com/cr/villanova/fullcit?p1432833.

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FENG, LIN. "MODULATION OF SYNAPTIC TRANSMISSION AT THE NUCLEUS TRACTUS SOLITARIUS." OpenSIUC, 2014. https://opensiuc.lib.siu.edu/dissertations/816.

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The caudal nucleus tractus solitarius (cNTS) is the key recipient of the primary afferents from visceral sensory neurons and also an important site that processes and integrates gastrointestinal, cardiovascular and respiratory functions. Glutamate and gamma-aminobutyric acid are the major neurotransmitters within the NTS, but studies have suggested that nicotinic acetylcholine receptors (nAChRs) and transient receptor potential (TRP) channels can modulate excitatory/inhibitory neurotransmission. I have designed studies to understand the role of nAChRs and TRP channels in the modulation of neurotransmission in the cNTS. In the first aim, experiments were designed to test the hypothesis that the cNTS contains function-specific subsets of neurons whose responsiveness to nicotine correlates with the target of their axonal projections. cNTS neurons send axonal projections to brain regions such as parabrachial nucleus (PBN), hypothalamic paraventricular nucleus (PVN), nucleus ambiguous (NA), dorsal motor nucleus of the vagus (DMV) and the caudal ventrolateral medulla (CVLM) and are involved in integrating autonomic and neuroendocrine functions. Presynaptic/postsynaptic modulation by nAChRs differ in the axonal projections of cNTS neurons, studying of which would provide better understanding of this complex integration. In vivo fluorescent tracing combined with in vitro slice patch-clamp electrophysiological recordings from anatomically identified caudal NTS neurons were used to study the expression and function of nAChRs (mainly á3â4 containing nAChRs) in the cNTS. Results from these studies demonstrate that presynaptic and postsynaptic responsiveness of caudal NTS neurons to nicotine correlates with the areas the neurons project to in the following order of prevalence: DMV>PVN>NA>CVLM>PBN (for presynaptic responses) and DMV>CVLM>PBN>NA>PVN (for postsynaptic responses). In the second aim, experiments were designed to test the hypothesis that nociceptive TRP channels TRPV1 (vanilloid) and TRPA1 (ankyrin) modulate synaptic transmission in the NTS. As a result of this modulation, the efferent functions that control autonomic and visceral functions will be regulated and account for the changes in autonomic neuropathy as patients with diabetes develop significant alterations in blood pressure and heart rate as well as silent myocardial ischemia as a result of blunted pain carrying ability. Results obtained from these studies demonstrated that TRPV1 and TRPA1 mRNA were detected in the dorsal root ganglion (DRG), but not in the NTS. Immunofluorescence studies revealed that TRPV1 and TRPA1 were expressed in the solitary tract central sensory terminals inputs to NTS but not in NTS neurons. This suggests that TRPV1 and TRPA1 are expressed only in solitary tract. Administration of capsaicin (TRPV1 agonist) and allyl isothiocyanate (AITC, TRPA1 agonist) both increased the frequency of s/mEPSCs without affecting spontaneous and miniature inhibitory postsynaptic currents (s/mIPSCs). Next, the modulation of TRPV1- and TRPA1-induced responses by utilizing a PKC activator (PDBu) was examined. Incubation of slices with PDBu synergistically increased the mEPSC frequency following capsaicin application suggesting an increased receptor affinity; however following application of AITC there was no significant change, suggesting that activation by covalent modification does not enhance binding affinity. Finally, the specificity of TRPV1 and TRPA1 effect on synaptic transmission by ablating TRPV1 and TRPA1were tested. There was no modulation of synaptic transmission in these animals, further confirming that capsaicin- and AITC-mediated modulation of synaptic transmission are specifically mediated by TRPV1 and TRPA1, respectively. Furthermore, animals with painful diabetic peripheral neuropthy exhibited enhanced synaptic activity at the NTS, suggesting a role in nociception and other visceral functions. In summary, nAChRs, TRPV1 and TRPA1 are expressed in the NTS and activation of which modulate excitatory synaptic transmission. The results obtained from these studies and their interpretation may provide a better understanding of the central mechanism of modulation on efferent functions from NTS that regulate cardiovascular, respiratory and gastrointestinal functions.

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D'Acunto, Mariantonietta. "Total synthesis of terpenoidic unsatured dialdehydes and evaluation of their activity towards TRP receptors." Doctoral thesis, Universita degli studi di Salerno, 2012. http://hdl.handle.net/10556/1764.

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2010 - 2011
The aim of this PhD project has been to develop new synthetic strategies in enantioselective preparation of natural products. In particular my attention has been focused on preparation of some natural metabolite, containing an α,β-unsaturated dialdehyde in a polycyclic backbone, and their synthetic analogue, in order to better understand structure activity relationship towards TRP receptors ion channels. The recent discover of the new thermoreceptor TRPA1 and given that these natural metabolites show also a widespread of bioactivities, such as antiproliferative and cytotoxic activity, has increased our interest towards these target ever more. Our purpose is to assay the bioactivity of synthesized products both as TRP receptor agonists and as antiproliferative compounds . The first chapter of this work is an introduction to these terpenoidic molecules, with a wide range of described natural occurring metabolite and their classification in drimane, isocopalane, and scalarane dialdehydes. Thus, the structure of TRP receptor is described with a brief history of these ion channels, starting from the first cloned receptor , the TRPV1 vanilloid. In the chapter 2 total syntheses of polygodial derivatives, both C-1 and C-3 functionalised, are described. Polygodial and C-1 functionalised drimanes have been prepared with an approach whose key step is a Diels Alder reaction; Drimane C-3 functionalised have been prepared with a radical chemistry approach... [edited by author]
X n.s.

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Stein, Marco Robert Philip. "Optochemical control of GABA(A) receptors and TRP channels and studies toward light-dependent regulation of NaV and mGluR6." Diss., Ludwig-Maximilians-Universität München, 2014. http://nbn-resolving.de/urn:nbn:de:bvb:19-181106.

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Colton, Craig K. "TRPV3 is a polymodal receptor." Columbus, Ohio : Ohio State University, 2006. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1164046830.

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Stein, Marco Robert Philip [Verfasser], and Dirk [Akademischer Betreuer] Trauner. "Optochemical control of GABA(A) receptors and TRP channels and studies toward light-dependent regulation of NaV and mGluR6 / Marco Robert Philip Stein. Betreuer: Dirk Trauner." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2014. http://d-nb.info/1069278661/34.

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Kim, Ju Young. "M1 muscarinic acetylcholine receptor regulation of endogenous transient receptor potential-canonical, subtype 6 (TRPC6) channels." Connect to resource, 2005. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1117570788.

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Thesis (Ph. D.)--Ohio State University, 2005.
Title from first page of PDF file. Document formatted into pages; contains xviii, 178 p.; also includes graphics. Includes bibliographical references (p. 163-178). Available online via OhioLINK's ETD Center

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Cao, De-Shou. "Role of transient receptor potential (TRP) channels in nociception /." Available to subscribers only, 2009. http://proquest.umi.com/pqdweb?did=1967913291&sid=2&Fmt=2&clientId=1509&RQT=309&VName=PQD.

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Cao, Deshou. "Role of Transient Receptor Potential (TRP) Channels in Nociception." OpenSIUC, 2009. https://opensiuc.lib.siu.edu/dissertations/71.

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Transient receptor potential (TRP) channels play an important role in sensory and nonsensory functions. TRPVanilloid 1 and TRPVanilloid 4 are proposed to be involved in inflammation-induced pain. TRPV1 is extensively studied and it is specifically involved in inflammatory thermal hypersensitivity. Mechanical hypersensitivity is one of the significant components of nociception. Several receptors have been proposed to underlie mechanosensation. The molecular entities responsible for mechanosensation are not fully understood. In this study, I have characterized the properties of TRPV4, a putative mechanosensitive ion channel expressed in dorsal root ganglion (DRG) neurons and nonsensory tissues. First, I have investigated the expression and function of TRPV4 and TRPV1 in the DRG neuronal cell bodies as well as their central terminals and determined the modulation by protein kinase C (PKC). Both TRPV4 and TRPV1 are expressed in DRG and laminae I and II of the spinal dorsal horn (DH). Ca2+ fluorescence imaging and whole-cell patch-clamp experiments showed that both capsaicin-induced TRPV1 response and 4alpha-phorbol 12, 13-didecanoate (4alpha-PDD)-induced TRPV4 response were observed in a proportion of the same DRG neurons, suggesting their co-expression. Incubation of DRG neurons with phorbol 12, 13-dibutyrate (PDBu), a PKC activator, resulted in a significantly greater potentiation of TRPV4 currents than TRPV1 currents. In HEK cells heterologously expressing TRPV4, PDBu potentiated TRPV4-mediated single-channel current activity. In patch-clamped DH neurons, the application of 4alpha-PDD at the first sensory synapse increased the frequency but not the amplitude of the miniature excitatory postsynaptic currents (mEPSCs), suggesting a presynaptic locus of action. 4alpha-PDD-induced increase in the frequency of mEPSC was further facilitated by PDBu. These results suggest that TRPV4 in the central terminals modulates synaptic transmission and is regulated by PKC. Second, I have studied the mechanosensitivity of TRPV4 in cell-attached patches by applying direct mechanical force via the patch pipette. In TRPV4 expressing HEK cells, the application of negative pressure evoked single-channel current activity in a reversible manner and the channel activity was enhanced after incubation with PDBu. TRPV4 has been shown to be activated by hypotonicity. Here I show that negative pressure exaggerated hypotonicity-induced single-channel current activity. However, in similar experimental conditions, cells expressing TRPV1 did not respond to mechanical force. TRP channels are also expressed in non-sensory regions and the role of these channels is not fully understood. Both TRPV4 and TRPV1 are expressed in the hippocampus. Using whole-cell patch-clamp techniques, I have found that 4alpha-PDD increased the frequency, but not the amplitude of mEPSCs in cultured hippocampal neurons, suggesting a presynaptic site of action. Interestingly, the application of capsaicin had no effect on synaptic transmission in hippocampal neuronal cultures. Finally, I have investigated the expression and function of TRP channels in diabetes because TRP channels have been shown to be involved in peripheral neuropathy as well as vascular complications in diabetes. ROS production plays a critical role in the progress of diabetes. I propose that lower levels of ROS up-regulate the expression TRP channels in the early stages of diabetes, leading to hyperalgesia, and higher levels of ROS or chronic exposure to ROS down-regulate TRP channels in the late stages of diabetes, resulting in hypoalgesia. I have found that the expression of TRPV1 and phospho p38 (p-p38) MAPK was increased in DRG of streptozotocin (STZ)-injected diabetic and non-diabetic hyperalgesic mice. An increase in TRPV1 and p-p38 MAPK levels was induced by STZ or H2O2 treatment in stably TRPV1 expressing HEK cells, suggesting the involvement of STZ-ROS-p38MAPK pathway. TRPV4 has been reported to be involved in vasodilatation by shear stress in blood vessels. Here, I have demonstrated that TRPV4 is expressed in lymphatic endothelial cells (LECs). Treatment with low concentration of H2O2 enhanced the expression of TRPV4 at mRNA and protein levels in LECs, suggesting that mild levels of ROS up-regulate TRPV4 expression. In diabetes, beta cell dysfunction is responsible for decreased insulin release. TRPV4 is expressed in RINm5F (beta cell line), islets and pancreas. It has been shown that hypotonicity induced insulin release in beta cell lines, which was mediated by activation of stretch-activated channels, raising the possibility of the involvement of TRPV4, a mechanosensitive channel. Therefore, I have studied the functional role of TRPV4 in beta cells. Incubation with 4alpha-PDD enhanced insulin release in RINm5F cells, suggesting TRPV4 regulates insulin secretion from pancreatic beta cells. Since TRPV4 expression levels are decreased in diabetes, insulin secretion from beta cells may be impaired. In summary, TRPV1, a thermosensitive channel, and TRPV4, a mechanosensitive channel, contribute to thermal and mechanical hyperalgesia, respectively in the early stage of DPN through their up-regulation by ROS-p38 MAPK and insulin/IGF-1 pathways. Due to the mechanical sensitivity of TRPV4 channel, the up-regulation in the early stage and down-regulation in the late stage may be involved in the development of vascular complications and regulation of insulin release in diabetes.

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Che, Hui, and 車慧. "Functional transient receptor potential channels in human preadipocytes and cardiac c-kit⁺ progenitor cells." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2013. http://hdl.handle.net/10722/196436.

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Transient receptor potential (TRP) channels play important roles in cellular physiology and biology. The present PhD project investigated the functional expression of TRPV and TRPM channels in human preadipocytes and cardiac c-kit+ progenitor cells and their roles in regulating cell proliferation, adipogenic differentiation or migration. In addition, the role of store-operated Ca2+ entry (SOCE) channels in regulating cell proliferation and migration was also studied in human cardiac c-kit+ progenitor cells using multiple approaches including whole-cell patch voltage-clamp, confocal microscope, molecular biology, etc. We found that TRPV2, TRPV4 and TRPM7 channels were abundantly expressed in human preadipocytes. Activation of TRPV2 channels by probenecid caused a long-lasting intracellular Ca2+ transient, while activation of TRPV4 channels by 4-PDD induced Ca2+ oscillations. TRPM7 current was recorded with a Mg2+-free pipette solution, and inhibited by 2-aminoethyl diphenyl borate (2-APB). Silence of TRPV2 or TRPM7, but not TRPV4, with the specific shRNA, reduced cell proliferation via inhibiting cyclin D1, cyclin E, and p-ERK1/2. Individually silencing these three channels decreased adipogenic differentiation by reducing p-Akt kinase. The results indicate that TRPV2, TRPV4 and TRPM7 are involved in adipogenesis, while TRPV2 and TRPM7, but not TRPV4, regulate cell proliferation in human preadipocytes. In second part of the thesis, abundant expression of TRPV2, TRPV4, and TRPM7 channels was demonstrated in human cardiac c-kit+ progenitor cells. Similar to human preadipocytes, probenecid and 4-PDD activated Ca2+ signaling, and TRPM7 current recorded with a Mg2+-free pipette solution was inhibited by 2-APB. Silencing TRPV2 or TRPM7, but not TRPV4, inhibited cell proliferation by arresting cells at G0/G1 phase with a reduced cyclin D, cyclin E, and p-ERK1/2. Cell migration was decreased with silence of TRPV2, TRV4 or TRPM7 via inhibiting p-Akt kinase. The results show that TRPV2, TRPV4 and TRPM7 mediate cell migration, while TRPV2 and TRPM7, but not TRPV4 channels, participate in regulating cell proliferation. In third part of the thesis, we demonstrated that SOCE channels were composed of TRPC1, STIM1 and Orai1 by protein-protein interaction. Silence of TRPC1, STIM1, or Orai1 with specific siRNA reduced Ca2+ influx through SOCE channels, decreased cell proliferation by inhibiting cyclin D1 and cyclin E, and slowed down cell migration via reducing p-Akt kinase. These results suggest that TRPC1, STIM1 and Orai1 are the major components of SOCE channels in human cardiac c-kit+ cells. SOCE channels play an essential role in regulating cell proliferation and migration. Collectively, this PhD project has demonstrated for the first time that 1) TRPV2, TRPV4, and TRPM7 are abundantly expressed in human preadipocytes and cardiac c-kit+ progenitor cells. 2) These TRP channels regulate adipogenic differentiation in preadipocytes and migration in cardiac c-kit+ progenitor cells. 3) TRPV2 and TRPM7, but not TRPV4, are involved in cell proliferation of human preadipocytes and cardiac c-kit+ progenitor cells. 4) TRPC1, STIM1 and Orai1 are interacted to form SOCE channels and regulate cell proliferation and migration in human cardiac c-kit+ cells. 5) All the above physiological roles of TRPV2, TRPV4, TRPM7, and SOCE channels are mediated by cyclin D1, cyclin E, p-ERK1/2, and/or p-Akt.
published_or_final_version
Medicine
Doctoral
Doctor of Philosophy

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Books on the topic "TRP receptors"

1

TRP ion channel function in sensory transduction and cellular signaling cascades. Boca Raton, FL: CRC/Taylor & Francis, 2007.

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J, Abramowitz, Flockerzi Veit, and Nilius B, eds. Transient receptor potential (TRP) channels. Berlin: Springer, 2007.

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Flockerzi, Veit, and Bernd Nilius, eds. Transient Receptor Potential (TRP) Channels. Berlin, Heidelberg: Springer Berlin Heidelberg, 2007. http://dx.doi.org/10.1007/978-3-540-34891-7.

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service), SpringerLink (Online, ed. Transient Receptor Potential Channels. Dordrecht: Springer Science+Business Media B.V., 2011.

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Nilius, Bernd, and Veit Flockerzi, eds. Mammalian Transient Receptor Potential (TRP) Cation Channels. Cham: Springer International Publishing, 2014. http://dx.doi.org/10.1007/978-3-319-05161-1.

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Nilius, Bernd, and Veit Flockerzi, eds. Mammalian Transient Receptor Potential (TRP) Cation Channels. Berlin, Heidelberg: Springer Berlin Heidelberg, 2014. http://dx.doi.org/10.1007/978-3-642-54215-2.

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Szallasi, Arpad. TRP channels in health and disease: Implications for diagnosis and therapy. Hauppauge, N.Y: Nova Science Publishers, 2010.

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Derek, Chadwick, Goode Jamie, and Novartis Foundation, eds. Mammalian TRP channels as molecular targets. Chichester: John Wiley, 2004.

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Gomtsyan, Arthur. Vanilloid receptor TRPV1 in drug discovery: Targeting pain and other pathological disorders. Hoboken, N.J: Wiley, 2010.

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Gomtsyan, Arthur. Vanilloid receptor TRPV1 in drug discovery: Targeting pain and other pathological disorders. Hoboken, N.J: Wiley, 2010.

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Book chapters on the topic "TRP receptors"

1

Trebak, M., L. Lemonnier, J. T. Smyth, G. Vazquez, and J. W. Putney. "Phospholipase C-Coupled Receptors and Activation of TRPC Channels." In Transient Receptor Potential (TRP) Channels, 593–614. Berlin, Heidelberg: Springer Berlin Heidelberg, 2007. http://dx.doi.org/10.1007/978-3-540-34891-7_35.

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Baez-Nieto, David, Juan Pablo Castillo, Constantino Dragicevic, Osvaldo Alvarez, and Ramon Latorre. "Thermo-TRP Channels: Biophysics of Polymodal Receptors." In Transient Receptor Potential Channels, 469–90. Dordrecht: Springer Netherlands, 2010. http://dx.doi.org/10.1007/978-94-007-0265-3_26.

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Zhu, Michael X., and Jisen Tang. "TRPC Channel Interactions with Calmodulin and IP3 Receptors." In Mammalian TRP Channels as Molecular Targets, 44–62. Chichester, UK: John Wiley & Sons, Ltd, 2008. http://dx.doi.org/10.1002/0470862580.ch4.

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Liu, Su, Shaozhen Xie, Yi-fen Lee, and Chawnshang Chang. "Physiological Functions of TR2 and TR4 Orphan Nuclear Receptor." In Nuclear Receptors, 327–43. Dordrecht: Springer Netherlands, 2010. http://dx.doi.org/10.1007/978-90-481-3303-1_13.

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Lehen’kyi, V’yacheslav, and Natalia Prevarskaya. "Oncogenic TRP Channels." In Transient Receptor Potential Channels, 929–45. Dordrecht: Springer Netherlands, 2010. http://dx.doi.org/10.1007/978-94-007-0265-3_48.

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Kaleta, Marta, and Christopher Palmer. "TRP Channels in Yeast." In Transient Receptor Potential Channels, 315–21. Dordrecht: Springer Netherlands, 2010. http://dx.doi.org/10.1007/978-94-007-0265-3_17.

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Xiao, Rui, and X. Z. Shawn Xu. "C. elegans TRP Channels." In Transient Receptor Potential Channels, 323–39. Dordrecht: Springer Netherlands, 2010. http://dx.doi.org/10.1007/978-94-007-0265-3_18.

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Wolstenholme, Adrian J., Sally M. Williamson, and Barbara J. Reaves. "TRP Channels in Parasites." In Transient Receptor Potential Channels, 359–71. Dordrecht: Springer Netherlands, 2010. http://dx.doi.org/10.1007/978-94-007-0265-3_20.

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Islam, Md Shahidul. "TRP Channels of Islets." In Transient Receptor Potential Channels, 811–30. Dordrecht: Springer Netherlands, 2010. http://dx.doi.org/10.1007/978-94-007-0265-3_42.

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Deinhardt, Katrin, and Moses V. Chao. "Trk Receptors." In Neurotrophic Factors, 103–19. Berlin, Heidelberg: Springer Berlin Heidelberg, 2014. http://dx.doi.org/10.1007/978-3-642-45106-5_5.

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Conference papers on the topic "TRP receptors"

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Timkin, Pavel, E. Timofeev, A. Chupalov, and Evgeniy Borodin. "ANALYSIS AND SELECTION OF LIGANDS FOR TRPM8 USING HARD DOCKING AND MACHINE LEARNING." In XIV International Scientific Conference "System Analysis in Medicine". Far Eastern Scientific Center of Physiology and Pathology of Respiration, 2020. http://dx.doi.org/10.12737/conferencearticle_5fe01d9b233509.17835494.

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In this work, using the in-silico experiment modeling method, the receptor and its ligands were docked in order to obtain the data necessary to study the possibility of using machine learning and hard intermolecular docking methods to predict potential ligands for various receptors. The protein TRPM8 was chosen, which is a member of the TRP superfamily of proteins and its classic agonist menthol as a ligand. It is known that menthol is able to bind to tyrosine 745 of the B chain. To carry out all the manipulations, we used the Autodock software and a special set of graphic tools designed to work with in silico models of chemicals. As a result of all the manipulations, the menthol conformations were obtained that can bind to the active center of the TRPM8 receptor.

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Huff, R. D., A. Robinson, M. H. Ryu, and C. Carlsten. "TRP and TLR receptors modulate airway responses to diesel exhaust and allergen co-exposure in sensitized participants." In ERS International Congress 2022 abstracts. European Respiratory Society, 2022. http://dx.doi.org/10.1183/13993003.congress-2022.4672.

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Miyata, T., T. Morita, T. Inomoto, S. Kawauchi, A. Shirakami, and S. Iwanaga. "PROTHROMBIN TOKUSHIMA: A REPLACEMENT OF ARGININE-418 BY TRYPTOPHAN THAT IMPAIRS THE FIBRINOGEN CLOTTING ACTIVITY OF DERIVED THROMBIN TOKUSHIMA." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643935.

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Recently, we identified a Japanese family having an abnormal prothrombin, designated “prothrombin Tokushima”. The propositus of this family is a 10-year-old female suffering from bleeding tendency. She is a double heterozygote transmitted from both the maternal dysprothrombinemia and the paternal hypoprothrombinemia. The purified prothrombin Tokushima exhibited the following properties: (i) factor Xa-catalyzed proteolysis of the abnormal prothrombin in the presence of factor Va, phospholipids and calcium ions yielded a two-chain thrombin, fragment 1 and fragment 2, (ii) thrombin derived from the abnormal molecule exhibited 21.5 % clotting activity relative to normal thrombin and reduced platelet aggregation activity, (iii) the apparent Km of thrombin Tokushima towards Boc-Val-Pro-Arg-MCA was 8-fold larger than that of normal thrombin, and the kcat value was approximately one-half of that of the normal enzyme, and (iv) its active site was fully titrable using P-nitrophenyl-P’-guanidinobenzoate.Subsequently, the structural studies on the abnormal thrombin were performed to identify the difference responsible for its reduced fibrinogen clotting activity upon conversion to thrombin. Amino acid sequence analysis of a peptide isolated from a lysyl endopeptidase digest of the abnormal thrombin indicated that Arg-418 (equivalent to Asn-101 in the chymotrypsin numbering system) had been replaced by Trp. This amino acid substitution can result from a single nucleotide change in the codon for Arg→4l8 (CGG→TGG). The Arg→Trp replacement found in the thrombin portion of prothrombin Tokushima appears to reduce its interaction with various substrates including fibrinogen and platelet receptors, and accounts for the recurrent bleeding episode observed in the propositus.

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Gao, Yandong, Dana Brantley-Sieders, Devi Majumdar, Jin Chen, Donna Webb, and Deyu Li. "A Simple Approach to Probe the Extracellular Signaling Pathways Using Ligand Traps." In ASME 2012 Third International Conference on Micro/Nanoscale Heat and Mass Transfer. American Society of Mechanical Engineers, 2012. http://dx.doi.org/10.1115/mnhmt2012-75106.

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Cells communicate with one another through a huge variety of extracellular soluble signaling molecules. A common method in biology to investigate the signaling pathways is to inactivate the gene coding the interested ligand or receptor from cells using modern DNA technology, known as gene knockout. Even though very effective, however, gene knockout is a time-consuming and cost-prohibitive process and requires huge amount of efforts to conduct. Here we present a simple method to probe the extracellular signaling pathways through engineering a semi-permeable barrier between two cell populations. In this approach, ligand traps, receptor-coated nano/micro-particles, are embedded inside the nanoporous barrier. Because the receptors have the ability to selectively bind to certain ligand(s) with high affinity, the associated ligands can be ‘trapped’ inside the barrier when they try to perfuse from one cell population to the other. As a result, the targeted soluble ligands can be effectively blocked from the molecular exchange between the two cell populations. We have demonstrated the feasibility of this novel approach using fluorescent proteins. An analytical model has also been developed to guide the design of the ligand-trap-embedded barrier.

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Fonseca, Anna Clara Silva, Nathália Brígida de Oliveira, and Daniela Camargos Costa. "PARTICIPAÇÃO DOS RECEPTORES TOLL-LIKE NA RESPOSTA A INFECÇÃO in vitro POR Leishmania (Viannia) braziliensis: UMA REVISÃO DE LITERATURA." In I Congresso Brasileiro de Parasitologia Humana On-line. Revista Multidisciplinar em Saúde, 2021. http://dx.doi.org/10.51161/rems/731.

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Introdução: A Leishmania braziliensis é um protozoário da família Trypanosomatidae responsável pelo desenvolvimento de uma das formas dermotrópicas da doença denominada leishmaniose mucosa (LM). O parasito é intracelular obrigatório e se multiplica dentro dos macrófagos e monócitos de seus hospedeiros. Os receptores tolllike (TLR) são glicoproteínas transmembrana presentes nas células de defesa, principalmente Natural Killer, macrófagos e células dendríticas, que reconhecem estruturas microbianas e que promovem uma série de sinais que produzem citocinas próinflamatórias importantes para que a resposta imune inata seja efetiva, como por exemplo TNF-α, IL-10 e TGF-β. Objetivo: Realizar revisão da literatura a respeito do papel dos receptores Toll-like na resposta à infecção in vitro por L. braziliensis. Material e métodos: Realizado no formato de revisão integrativa, a pesquisa abrangeu artigos científicos das bases de dados SCIELO e PUBMED. Os descritores utilizados foram “American Cutaneous leishmaniasis”, “Leishmania braziliensis”, “mucosal leishmaniasis” e “Toll-like receptors”. Foram selecionados artigos acadêmicos originais, escritos na língua inglesa, que foram publicados no período de 2014 a 2020, obtendo -se 20 artigos. Após a leitura dos títulos e resumos e ao adotar os critérios de exclusão, foram selecionados 12 artigos originais para a presente revisão. Resultados: As pesquisas selecionadas avaliaram 147 pacientes com diagnóstico histopatológico de Leishmaniose Cutânea (LC). Os pacientes que apresentavam LM, concomitantemente possuíam uma maior expressão de células TCD4+, TCD8+, IL-10+ e TGF-β+ em comparação com aqueles que possuíam LC, que expressavam uma menor quantidades dessas citocinas. Em oposição, o TNF-α em pacientes com LC, encontrava-se aumentado se comparada a LM. Na análise relacionada a TLR-2 e TLR-4, houve uma maior expressão em monócitos nos pacientes com LC associadas a L. braziliensis e que o bloqueio dos TRL reduziu as respostas oxidativas das células de pacientes com LC. Conclusão: Diante dos resultados é possível concluir que a maior expressão de TLR-2 e TLR-4 promove a hiper-reatividade de citocinas pró-inflamatórias e consequente pré-disposição para desenvolvimento da doença. Portanto, seu bloqueio é eficaz para redução das lesões. Esse dado demonstra a relação e o papel dos TLR na infecção por L. braziliensis, trazendo importantes esclarecimentos para a resposta imune e desenho vacinal.

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Brewer, Bryson M., Yandong Gao, Rebecca M. Sappington, and Deyu Li. "Microfluidic Molecular Trap: Probing Extracellular Signaling by Selectively Blocking Exchange of Specific Molecules in Cell-Cell Interactions." In ASME 2013 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2013. http://dx.doi.org/10.1115/imece2013-64489.

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Communication among cell populations is achieved via a wide variety of soluble, extracellular signaling molecules [1]. In order to investigate the role of specific molecules in a cellular process, researchers often utilize in vitro cell culture techniques in which the molecule under question has been removed from the signaling pathway. Traditionally, this has been accomplished by eliminating the gene in the cell that is responsible for coding the targeted ligand/receptor by using modern DNA technology such as gene knockout; however, this process is expensive, time-consuming, and labor intensive. Previously, we have demonstrated a microfluidic platform that uses a semi-permeable barrier with embedded receptor-coated nanoparticles to selectively remove a specific molecule or ligand from the extracellular signaling pathway in a cell co-culture environment [2]. This initial proof-of-principle was conducted using biotinylated nanoparticles and fluorescently tagged avidin molecules, as the avidin/biotin complex is the strongest known non-covalent interaction between a protein and a ligand (Dissociation constant kd = 10−15 M). Also, the trap was only effective for short time periods (<15 min) because the high concentration of fluorescently tagged avidin molecules required for visualization quickly saturated the barrier. However, nearly all biologically relevant ligand-receptor interactions have lower binding affinities than the avidin-biotin complex, with dissociation constants that are larger by several orders of magnitude. In addition, many in vitro cell culture experiments are conducted over multiple hours or days. Thus, a practically useful molecular trap device must be able to operate in a lower binding affinity regime while also lasting for extended time periods. Here we present results in which a biotinylated-particle barrier was used to successfully block lower concentrations of fluorescently tagged avidin for multiple days, showcasing the applicability of the device for long term experiments. In addition, we introduce a modified molecular trap in which the protein A/goat IgG complex was used to demonstrate the effectiveness of the platform for lower binding affinity protein-ligand interactions. These results indicate the potential usefulness of the microfluidic molecular trap platform for probing extracellular signaling pathways.

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Bonvini, Sara, Eric Dubuis, John Adcock, Michael Wortley, Mark Birrell, and Maria Belvisi. "Activation of transient receptor potential (TRP) channels by hypoosmolar solution: an endogenous mechanism of ATP release and afferent nerve activation." In ERS International Congress 2017 abstracts. European Respiratory Society, 2017. http://dx.doi.org/10.1183/1393003.congress-2017.oa4410.

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Kishimoto, Tatsunori, Yasuyo Maezawa, Suguru N. Kudoh, Takahisa Taguchi, and Chie Hosokawa. "Molecular dynamics in an optical trap of glutamate receptors labeled with quantum-dots on living neurons." In SPIE Technologies and Applications of Structured Light, edited by Takashige Omatsu. SPIE, 2017. http://dx.doi.org/10.1117/12.2275026.

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Champaigne, Kevin D., Sarette N. Jenderny, and Jiro Nagatomi. "Electrophysiological Investigation of Hydrostatic Pressure Mechanotransduction by Urothelial Cell Lines." In ASME 2011 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2011. http://dx.doi.org/10.1115/sbc2011-53518.

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The urothelium is the epithelial lining of the ureters, urinary bladder, and urethra. Recent discoveries have suggested that in addition to providing a barrier function to urine, the urothelium actively participates in sensory functions related to thermal, chemical, and mechanical stimuli, and releases chemical signals in response[1]. In addition to a sensitivity to cell membrane stretch caused by wall tension upon bladder filling, in vitro studies by our group have shown that urothelial cells may be sensitive to hydrostatic pressure directly without requiring membrane stretching [2]. Specifically, primary cultures of rat bladder urothelial cells exposed to 10 cmH2O pressure on rigid substrates released significantly greater amounts of ATP compared to the baseline control without exposure to pressure. Moreover, this ATP response by rat urothelial cells to pressure was inhibited by pre-treatment of cells with ruthenium red, a non-specific antagonist of transient receptor potential (TRP) channels, suggesting a potential involvement of these channels in pressure mechanotransduction. Further understanding of the mechanisms, however, is needed to improve treatment of bladder dysfunction such as overactive bladder.

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Olsen, Shawn, and Jiro Nagatomi. "An Optical Approach for Studying the Cellular Mechanotransduction of Hydrostatic Pressure by Bladder Urothelial Cells." In ASME 2011 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2011. http://dx.doi.org/10.1115/sbc2011-53872.

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Recent studies have suggested that the bladder urothelium is sensitive to both stretch and hydrostatic pressure during bladder filling, and is considered to play a mechanosensory role in sensing bladder fullness [1, 2]. In a previous study [3], our group demonstrated that compared to the control, rat bladder urothelial cells (UCs) exposed to hydrostatic pressure (10–15 cmH2O for 5 minutes) in vitro released significantly higher levels of ATP and that this response was attenuated by pharmacologically blocking transient receptor potential (TRP) channels, as well as epithelial sodium channels (ENACs). While blocking these ion channels inhibited the ATP response by UCs to hydrostatic pressure, it remains unclear whether these ion channels are being activated directly by hydrostatic pressure or by membrane deformation. Our current hypothesis is that a change in cell volume may occur due to the application of hydrostatic pressure and subsequent changes in cellular osmolality, which, in turn, activate the membrane-bound mechanosensitive channels. Using real-time fluorescent imaging and a custom experimental setup, the present study sought to quantify the UC cell volume changes during exposure to hydrostatic pressure and to better understand the mechanisms by which UCs sense hydrostatic pressure.

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Reports on the topic "TRP receptors"

1

Webster, Nicholas J., Stan Krajewski, and Michael C. Pirrung. Small Molecule Activators of the TRK Receptors for Neuroprotection. Fort Belvoir, VA: Defense Technical Information Center, May 2011. http://dx.doi.org/10.21236/ada555936.

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Zhang, Xiao-kun. Orphan Receptor TR3/nur77 and Apoptosis in Prostate Cancer Cells. Fort Belvoir, VA: Defense Technical Information Center, July 2002. http://dx.doi.org/10.21236/ada410922.

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Spiegelman, Vladimir S. The Role of Beta-TrCP Ubiquitin Ligase Receptor in the Development of Breast Cancer. Fort Belvoir, VA: Defense Technical Information Center, June 2006. http://dx.doi.org/10.21236/ada484616.

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Funkenstein, Bruria, and Cunming Duan. GH-IGF Axis in Sparus aurata: Possible Applications to Genetic Selection. United States Department of Agriculture, November 2000. http://dx.doi.org/10.32747/2000.7580665.bard.

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Many factors affect growth rate in fish: environmental, nutritional, genetics and endogenous (physiological) factors. Endogenous control of growth is very complex and many hormone systems are involved. Nevertheless, it is well accepted that growth hormone (GH) plays a major role in stimulating somatic growth. Although it is now clear that most, if not all, components of the GH-IGF axis exist in fish, we are still far from understanding how fish grow. In our project we used as the experimental system a marine fish, the gilthead sea bream (Sparus aurata), which inhabits lagoons along the Mediterranean and Atlantic coasts of Europe, and represents one of the most important fish species used in the mariculture industry in the Mediterranean region, including Israel. Production of Sparus is rapidly growing, however, in order for this production to stay competitive, the farming of this fish species has to intensify and become more efficient. One drawback, still, in Sparus extensive culture is that it grows relatively slow. In addition, it is now clear that growth and reproduction are physiological interrelated processes that affect each other. In particular sexual maturation (puberty) is known to be closely related to growth rate in fish as it is in mammals, indicating interactions between the somatotropic and gonadotropic axes. The goal of our project was to try to identify the rate-limiting components(s) in Sparus aurata GH-IGF system which might explain its slow growth by studying the ontogeny of growth-related genes: GH, GH receptor, IGF-I, IGF-II, IGF receptor, IGF-binding proteins (IGFBPs) and Pit-1 during early stages of development of Sparus aurata larvae from slow and fast growing lines. Our project was a continuation of a previous BARD project and could be divided into five major parts: i) obtaining additional tools to those obtained in the previous project that are necessary to carry out the developmental study; ii) the developmental expression of growth-related genes and their cellular localization; iii) tissue-specific expression and effect of GH on expression of growth-related genes; iv) possible relationship between GH gene structure, growth rate and genetic selection; v) the possible role of the IGF system in gonadal development. The major findings of our research can be summarized as follows: 1) The cDNAs (complete or partial) coding for Sparus IGFBP-2, GH receptor and Pit-1 were cloned. Sequence comparison reveals that the primary structure of IGFBP-2 protein is 43-49% identical to that of zebrafish and other vertebrates. Intensive efforts resulted in cloning a fragment of 138 nucleotides, coding for 46 amino acids in the proximal end of the intracellular domain of GH receptor. This is the first fish GH receptor cDNA that had been cloned to date. The cloned fragment will enable us to complete the GH - receptor cloning. 2) IGF-I, IGF-II, IGFBP-2, and IGF receptor transcripts were detected by RT-PCR method throughout development in unfertilized eggs, embryos, and larvae suggesting that these mRNAs are products of both the maternal and the embryonic genomes. Preliminary RT-PCR analysis suggest that GH receptor transcript is present in post-hatching larvae already on day 1. 3) IGF-1R transcripts were detected in all tissues tested by RT-PCR with highest levels in gill cartilage, skin, kidney, heart, pyloric caeca, and brain. Northern blot analysis detected IGF receptor only in gonads, brain and gill cartilage but not in muscle; GH increased slightly brain and gill cartilage IGF-1R mRNA levels. 4) IGFBP-2 transcript were detected only in liver and gonads, when analyzed by Northern blots; RT-PCR analysis revealed expression in all tissues studied, with the highest levels found in liver, skin, gonad and pyloric caeca. 5) Expression of IGF-I, IGF-II, IGF-1R and IGFBP-2 was analyzed during gonadal development. High levels of IGF-I and IGFBP-2 expression were found in bisexual young gonads, which decreased during gonadal development. Regardless of maturational stage, IGF-II levels were higher than those of IGF-L 6) The GH gene was cloned and its structure was characterized. It contains minisatellites of tandem repeats in the first and third introns that result in high level of genetic polymorphism. 7) Analysis of the presence of IGF-I and two types of IGF receptor by immunohistochemistry revealed tissue- and stage-specific expression during larval development. Immunohistochemistry also showed that IGF-I and its receptors are present in both testicular and ovarian cells. Although at this stage we are not able to pinpoint which is the rate-limiting step causing the slow growth of Sparus aurata, our project (together with the previous BARD) yielded a great number of experimental tools both DNA probes and antibodies that will enable further studies on the factors regulating growth in Sparus aurata. Our expression studies and cellular localization shed new light on the tissue and developmental expression of growth-related genes in fish.

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J. Bland. Calculation: Values and Consumption Rates of Locally Produced Food and Tap Water for the Receptor of Interest. Office of Scientific and Technical Information (OSTI), July 2000. http://dx.doi.org/10.2172/894058.

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Eyal, Yoram, and Sheila McCormick. Molecular Mechanisms of Pollen-Pistil Interactions in Interspecific Crossing Barriers in the Tomato Family. United States Department of Agriculture, May 2000. http://dx.doi.org/10.32747/2000.7573076.bard.

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During the evolutionary process of speciation in plants, naturally occurring barriers to reproduction have developed that affect the transfer of genes within and between related species. These barriers can occur at several different levels beginning with pollination-barriers and ending with hybrid-breakdown. The interaction between pollen and pistils presents one of the major barriers to intra- and inter-specific crosses and is the focus of this research project. Our long-term goal in this research proposal was defined to resolve questions on recognition and communication during pollen-pistil interactions in the extended tomato family. In this context, this work was initiated and planned to study the potential involvement of tomato pollen-specific receptor-like kinases (RLK's) in the interaction between pollen and pistils. By special permission from BARD the objectives of this research were extended to include studies on pollen-pistil interactions and pollination barriers in horticultural crops with an emphasis on citrus. Functional characterization of 2 pollen-specific RLK's from tomato was carried out. The data shows that both encode functional kinases that were active as recombinant proteins. One of the kinases was shown to accumulate mainly after pollen germination and to be phosphorylated in-vitro in pollen membranes as well as in-vivo. The presence of style extract resulted in dephosphorylation of the RLK, although no species specificity was observed. This data implies a role for at least one RLK in pollination events following pollen germination. However, a transgenic plant analysis of the RLK's comprising overexpression, dominant-negative and anti-sense constructs failed to provide answers on their role in pollination. While genetic effects on some of the plants were observed in both the Israeli and American labs, no clear functional answers were obtained. An alternative approach to addressing function was pursued by screening for an artificial ligand for the receptor domain using a peptide phage display library. An enriched peptide sequence was obtained and will be used to design a peptide-ligand to be tested for its effect o pollen germination and tube growth. Self-incompatibility (SI) in citrus was studied on 3 varieties of pummelo. SI was observed using fluorescence microscopy in each of the 3 varieties and compatibility relations between varieties was determined. An initial screen for an S-RNase SI mechanism yielded only a cDNA homologous to the group of S-like RNases, suggesting that SI results from an as yet unknown mechanism. 2D gel electrophoresis was applied to compare pollen and style profiles of different compatibility groups. A "polymorphic" protein band from style extracts was observed, isolated and micro-sequenced. Degenerate primers designed based on the peptide sequence date will be used to isolate the relevant genes i order to study their potential involvement in SI. A study on SI in the apple cultivar Top red was initiated. SI was found, as previously shown, to be complete thus requiring a compatible pollinator variety. A new S-RNase allele was discovered fro Top red styles and was found to be highly homologous to pear S-RNases, suggesting that evolution of these genes pre-dated speciation into apples and pears but not to other Rosaceae species. The new allele provides molecular-genetic tools to determine potential pollinators for the variety Top red as well as a tool to break-down SI in this important variety.

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Naim, Michael, Andrew Spielman, Shlomo Nir, and Ann Noble. Bitter Taste Transduction: Cellular Pathways, Inhibition and Implications for Human Acceptance of Agricultural Food Products. United States Department of Agriculture, February 2000. http://dx.doi.org/10.32747/2000.7695839.bard.

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Historically, the aversive response of humans and other mammals to bitter-taste substances has been useful for survival, since many toxic constituents taste bitter. Today, the range of foods available is more diverse. Many bitter foods are not only safe for consumption but contain bitter constituents that provide nutritional benefits. Despite this, these foods are often eliminated from our current diets because of their unacceptable bitterness. Extensive technology has been developed to remove or mask bitterness in foods, but a lack of understanding of the mechanisms of bitterness perception at the taste receptor level has prevented the development of inhibitors or efficient methods for reducing bitterness. In our original application we proposed to: (a) investigate the time course and effect of selected bitter tastants relevant to agricultural products on the formation of intracellular signal molecules (cAMP, IP3, Ca2+) in intact taste cells, in model cells and in membranes derived therefrom; (b) study the effect of specific bitter taste inhibitors on messenger formation and identify G-proteins that may be involved in tastant-induced bitter sensation; (c) investigate interactions and self-aggregation of bitter tastants within membranes; (d) study human sensory responses over time to these bitter-taste stimuli and inhibitors in order to validate the biochemical data. Quench-flow module (QFM) and fast pipetting system (FPS) allowed us to monitor fast release of the aforementioned signal molecules (cGMP, as a putative initial signal was substituted for Ca2+ ions) - using taste membranes and intact taste cells in a time range below 500 ms (real time of taste sensation) - in response to bitter-taste stimulation. Limonin (citrus) and catechin (wine) were found to reduce cellular cAMP and increase IP3 contents. Naringin (citrus) stimulated an IP3 increase whereas the cheese-derived bitter peptide cyclo(leu-Trp) reduced IP3 but significantly increased cAMP levels. Thus, specific transduction pathways were identified, the results support the notion of multiple transduction pathways for bitter taste and cross-talk between a few of those transduction pathways. Furthermore, amphipathic tastants permeate rapidly (within seconds) into liposomes and taste cells suggesting their availability for direct activation of signal transduction components by means of receptor-independent mechanisms within the time course of taste sensation. The activation of pigment movement and transduction pathways in frog melanophores by these tastants supports such mechanisms. Some bitter tastants, due to their amphipathic properties, permeated (or interacted with) into a bitter tastant inhibitor (specific phospholipid mixture) which apparently forms micelles. Thus, a mechanism via which this bitter taste inhibitor acts is proposed. Human sensory evaluation experiments humans performed according to their 6-n-propyl thiouracil (PROP) status (non-tasters, tasters, super-tasters), indicated differential perception of bitterness threshold and intensity of these bitter compounds by different individuals independent of PROP status. This suggests that natural products containing bitter compounds (e.g., naringin and limonin in citrus), are perceived very differently, and are in line with multiple transduction pathways suggested in the biochemical experiments. This project provides the first comprehensive effort to explore the molecular basis of bitter taste at the taste-cell level induced by economically important and agriculturally relevant food products. The findings, proposing a mechanism for bitter-taste inhibition by a bitter taste inhibitor (made up of food components) pave the way for the development of new, and perhaps more potent bitter-taste inhibitors which may eventually become economically relevant.

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Ullman, Diane E., Benjamin Raccah, John Sherwood, Meir Klein, Yehezkiel Antignus, and Abed Gera. Tomato Spotted Wilt Tosporvirus and its Thrips Vectors: Epidemiology, Insect/Virus Interactions and Control. United States Department of Agriculture, November 1999. http://dx.doi.org/10.32747/1999.7573062.bard.

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Objectives. The major aim of the proposed research was to study thrips-TSWV relationships and their role in the epidemiology of the virus with the aim of using this knowledge to reduce crop losses occurring due to epidemics. Our specific objectives were: To determine the major factors involved in virus outbreaks, including: a) identifying the thrips species involved in virus dissemination and their relative role in virus spread; b) determining the virus sources among wild and cultivated plants throughout the season and their role in virus spread, and, c) determining how temperature and molecular variations in isolates impact virus replication in plants and insects and impact the transmission cycle. Background to the topic. Tospoviruses are among the most important emerging plant viruses that impact production of agricultural and ornamental crops. Evolution of tospoviruses and their relationships with thrips vector species have been of great interest because of crop damage caused world wide and the complete absence of suitable methods of control. Tospoviruses threaten crops in Israel and the United States. By understanding the factors contributing to epidemics and the specific relationships between thrips species and particular tospoviruses we hope that new strategies for control can be developed that will benefit agriculture in both Israel and the United States. Major conclusions, solutions, achievements. We determined that at least three tospoviruses were involved in epidemics in Israel and the United States, tomato spotted wilt virus (TSWV), impatiens necrotic spot virus (INSV) and iris yellow spot virus (IYSV). We detected and characterized INSV for the first time in Israel and, through our efforts, IYSV was detected and characterized for the first time in both countries. We demonstrated that many thrips species were present in commercial production areas and trap color influenced thrips catch. Frankliniella occidentalis was the major vector species of INSV and TSWV and populations varied in transmission efficiency. Thrips tabaci is the sole known vector of IYSV and experiments in both countries indicated that F. occidentalis is not a vector of this new tospovirus. Alternate plant hosts were identified for each virus. A new monitoring system combining sticky cards and petunia indicator plants was developed to identify sources of infective thrips. This system has been highly successful in the U.S. and was used to demonstrate to growers that removal of plant sources of infective thrips has a dramatic impact on virus incidence. Finally, a putative thrips receptor mediating acquisition of TSWV was discovered. Implications, scientific and agricultural. Our findings have contributed to new control measures that will benefit agriculture. Identification of a putative thrips receptor for TSWV and our findings relative to thrips/tospovirus specificity have implications for development of innovative new control strategies.

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Gothilf, Yoav, Roger Cone, Berta Levavi-Sivan, and Sheenan Harpaz. Genetic manipulations of MC4R for increased growth and feed efficiency in fish. United States Department of Agriculture, January 2016. http://dx.doi.org/10.32747/2016.7600043.bard.

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The hypothalamic melanocortin system plays a central role in the regulation of food consumption and energy homeostasis in mammals. Accordingly, our working hypothesis in this project was that genetic editing of the mc4r gene, encoding Melanocortin Receptor 4 (MC4R), will enhance food consumption, feed efficiency and growth in fish. To test this hypothesis and to assess the utility of mc4r editing for the enhancement of feed efficiency and growth in fish, the following objectives were set: Test the effect of the mc4r-null allele on feeding behavior, growth, metabolism and survival in zebrafish. Generate mc4r-null alleles in tilapia and examine the consequences for growth and survival, feed efficiency and body composition. Generate and examine the effect of naturally-occurring mc4r alleles found in swordfish on feeding behavior, growth and survival in zebrafish. Define the MC4R-mediated and MC4R-independent effects of AgRP by crossing mc4r- null strains with fish lacking AgRP neurons or the agrpgene. Our results in zebrafish did not support our hypothesis. While knockout of the agrpgene or genetic ablation of hypothalamic AgRP neurons led to reduced food intake in zebrafish larvae, knockout (KO) of the mc4r gene not only did not increase the rate of food intake but even reduced it. Since Melanocortin Receptor 3 (MC3R) has also been proposed to be involved in hypothalamic control of food intake, we also tested the effectofmc3r gene KO. Again, contrary to our hypothesis, the rate of food intake decreased. The next step was to generate a double mutant lucking both functional MC3R and MC4R. Again, the double KO exhibited reduced food intake. Thus, the only manipulation within the melanocortin system that affected food intake in consistent with the expected role of the system was seen in zebrafish larvae upon agrpKO. Interestingly, despite the apparent reduced food intake in the larval stage, these fish grow to be of the same size as wildtype fish at the adult stage. Altogether, it seems that there is a compensatory mechanism that overrides the effect of genetic manipulations of the melanocortin system in zebrafish. Under Aim 3, we introduced the Xna1, XnB1l, and XnB2A mutations from the Xiphophorus MC4R alleles into the zebrafish MC4R gene. We hypothesized that these MC4R mutations would act as dominant negative alleles to increase growth by suppressing endogenous MC4R activity. When we examined the activity of the three mutant alleles, we were unable to document any inhibition of a co-transfected wild type MC4R allele, hence we did not introduce these alleles into zebrafish. Since teleost fish possess two agrpgenes we also tested the effect of KO of the agrp2 gene and ablation of the AgRP2 cells. We found that the AgRP2 system does not affect food consumption but may rather be involved in modulating the stress response. To try to apply genetic editing in farmed fish species we turned to tilapia. Injection of exogenous AgRP in adult tilapia induced significant changes in the expression of pituitary hormones. Genetic editing in tilapia is far more complicated than in zebrafish. Nevertheless, we managed to generate one mutant fish carrying a mutation in mc4r. That individual died before reaching sexual maturity. Thus, our attempt to generate an mc4r-mutant tilapia line was almost successful and indicate out non-obvious capability to generate mutant tilapia.

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