Дисертації з теми "Kidney epithelial cells"
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1
Tang, Chi-wai Sydney. "The many facets of the renal proximal tubular epithelial cell in human." Click to view the E-thesis via HKUTO, 2005. http://sunzi.lib.hku.hk/hkuto/record/B31992468.
2
Tang, Chi-wai Sydney, and 鄧智偉. "The many facets of the renal proximal tubular epithelial cell inhuman." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2005. http://hub.hku.hk/bib/B31992468.
3
Measures, H. R. "A study of desmosome formation in kidney epithelial cells." Thesis, University of Southampton, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.234435.
4
Xie, Jianxun. "Involvement of transcription factors in cadmium-induced apoptosis and cell cycle arrest in rat kidney cells /." View online ; access limited to URI, 2005. http://0-wwwlib.umi.com.helin.uri.edu/dissertations/dlnow/3206258.
5
Lim, Ai Ing, and 林艾盈. "Shedding of kidney injury molecule-1 by kidney proximal tubular epithelial cells: the role of matrixmetalloproteinase-3." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2012. http://hub.hku.hk/bib/B49799745.
Анотація:
Regardless of the original cause and etiology, the progression of kidney disease follows a final common pathway associated with tubulointerstitial injury, in which proximal tubular epithelial cells (PTEC) are instrumental. Kidney injury molecule-1 (KIM-1) is an emerging biomarker of kidney tubular damage. It is markedly expressed and released into urine in various animal models and human kidney diseases. This study aimed to explore the underlying mechanism regulating the release of KIM-1 by PTEC.
First, expression and release of KIM-1 by primary cultured human PTEC were examined. In quiescent PTEC, KIM-1 was detected at the plasma membrane and in the cytoplasm. A transwell system, in which PTEC were grown as monolayer on permeable membrane, was used to examine the polarized release of KIM-1. PTEC constitutively released KIM-1 from their apical surface, and the release was independent of gene expression or protein synthesis. The KIM-1 release process by PTEC was enhanced dose- and time-dependently by two important kidney injury mediators, human serum albumin (HSA) and tumor necrosis factor (TNF)-α, and was inhibited by the presence of broad-spectrum inhibitors of matrix metalloproteinases (MMP).
Second, the potential sheddases responsible for KIM-1 shedding were identified by quantitative polymerase chain reaction (PCR) array system, in which the gene expression of a panel of MMP members was screened. The gene expression of MMP-3, MMP-7 and MMP-9 was up-regulated by PTEC under HSA or TNF-α activation. Blockade experiments with synthetic MMP inhibitors or MMP gene knockdown by small interfering RNA transfection, revealed that the constitutive or accelerated KIM-1 shedding was mediated by MMP-3, but not MMP-7 or MMP-9. The role of MMP-3 in KIM-1 shedding was further defined by additional data showing the enhanced MMP-3 synthesis by HSA- or TNF-α-stimulated PTEC, and the up-regulated KIM-1 shedding by PTEC following exogenous MMP-3 treatment.
Third, the regulatory mechanism of MMP-3-mediated KIM-1 shedding was investigated. Treatment of PTEC with HSA or TNF-α up-regulated the reactive oxygen species (ROS) generation, and its kinetics ran parallel to the increase of KIM-1 shedding and MMP-3 synthesis. In addition, exogenous hydrogen peroxide dose-dependently induced KIM-1 shedding and MMP-3 synthesis, which were abolished by the presence of an oxidation inhibitor. These evidence suggest that ROS play an essential role in regulating the MMP-3-mediated KIM-1 shedding by PTEC.
Finally, a mouse model of acute kidney injury induced by renal ischemia and reperfusion (I/R) was established to translate the in vitro findings. Reduced kidney function and increased urinary KIM-1 level were observed in mice after renal I/R treatment. Strikingly, the expression of MMP-3 and KIM-1 in the I/R treated mice was most profound in the S3 segments of the proximal tubules, where is the most susceptible area to oxidative stress. Taken together, these in vivo data have further strengthened the distinct roles of ROS and MMP-3 in KIM-1 shedding during PTEC injury.
In conclusion, ROS generated by the injured PTEC activate MMP-3, which release the soluble KIM-1 through the ectodomain shedding process.published_or_final_version
Medicine
Master
Master of Philosophy
6
Zhou, Li. "The molecular mechanisms of aristolochic acid nephropathy." Click to view the E-thesis via HKUTO, 2009. http://sunzi.lib.hku.hk/hkuto/record/B43224349.
7
Laestadius, Åsa. "Cellular mechanisms of interaction between uropathogenic Escherichia coli and renal epithelial cells /." Stockholm, 2002. http://diss.kib.ki.se/2002/91-7349-187-X.
8
Sampangi, Sandeep. "Autologous human kidney proximal tubule epithelial cells (PTEC) modulate dendritic cell (DC), T cell and B cell responses." Thesis, Queensland University of Technology, 2015. https://eprints.qut.edu.au/82033/1/Sandeep_Sampangi_Thesis.pdf.
Анотація:
This is a comprehensive study of human kidney proximal tubular epithelial cells (PTEC) which are known to respond to and mediate the pathological process of a range of kidney diseases. It identifies various molecules expressed by PTEC and how these molecules participate in down-regulating the inflammatory process, thereby highlighting the clinical potential of these molecules to treat various kidney diseases. In the disease state, PTEC gain the ability to regulate the immune cell responses present within the interstitium. This down-regulation is a complex interaction of contact dependent/independent mechanisms involving various immuno-regulatory molecules including PD-L1, sHLA-G and IDO. The overall outcome of this down-regulation is suppressed DC maturation, decreased number of antibody producing B cells and low T cell responses. These manifestations within a clinical setting are expected to dampen the ongoing inflammation, preventing the damage caused to the kidney tissue.
9
Broadbelt, Nalini V. "Regulation of iNOS expression : in response to pressure in proximal tubule epithelial cells /." Access full-text from WCMC, 2008. http://proquest.umi.com/pqdweb?did=1619205731&sid=2&Fmt=2&clientId=8424&RQT=309&VName=PQD.
10
Miskovic, Dragana. "A characterization of BiP gene expression in Xenopus laevis embryos and A6 kidney epithelial cells." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape7/PQDD_0021/NQ38257.pdf.
11
Söderblom, Tomas. "Effects of bacterial toxins on calcium homeostasis in renal inflammation /." Stockholm, 2004. http://diss.kib.ki.se/2004/91-7349-904-8/.
12
Olteanu, Dragos S. "Dysregulated ENAC and NHE function in cilium-deficient renal collecting duct cell monolayers a model of polycystic kidney disease /." Thesis, Birmingham, Ala. : University of Alabama at Birmingham, 2007. https://www.mhsl.uab.edu/dt/2009r/olteanu.pdf.
13
Zhou, Li, and 周莉. "The molecular mechanisms of aristolochic acid nephropathy." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2009. http://hub.hku.hk/bib/B43224349.
14
Law, Becker M. P. "The functional characterisation of human innate lymphocytes in renal fibrosis and chronic kidney disease." Thesis, Queensland University of Technology, 2019. https://eprints.qut.edu.au/132513/1/Becker%20Meng-Po_Law_Thesis.pdf.
Анотація:
This thesis by publication is a step forward in understanding the function of discrete immune cell populations in kidneys with chronic inflammation and fibrosis. We have successfully identified various human immune cells of the innate immune system as critical drivers of chronic kidney disease. The findings of this thesis sheds light on novel functions of innate immune cells and opens opportunities for the development of novel kidney therapies.
15
Olsson, Magnus. "Nuclear pore membrane glycoprotein 210 as a new marker for epithelial cells." Doctoral thesis, Uppsala University, Department of Cell and Molecular Biology, 2003. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-3265.
Анотація:
Epithelial cell polarisation is a prerequisite for the branching morphogenesis in several organs. Differential screening techniques were used to identify genes, which are upregulated during induction of epithelium in early kidney development. This investigation revealed two separate genes, Nuclear localising protein 1 (Nulp1), a previously undescribed gene with sequence characteristics of the basic helix-loop-helix transcription factor family, and glycoprotein 210 (gp210, POM210), an integral membrane protein constituent of the nuclear pore complex (NPC). Of these, gp210 was found to be upreglated during conversion of mesenchyme to epithelium.
The nuclear envelope, which demarcates the nuclear region in the eukaryotic cell, consists of an inner and an outer membrane that are fused at the locations for NPCs. These large macromolecular assemblages are tube like structures connecting the cytoplasmic and nuclear compartments of the cell. NPCs serve as the only conduits for exchange of molecular information between these cellular rooms. Electron microscopy techniques have revealed detailed information about the NPC architecture. A number of proteins (nucleoporins) have been characterised and embodied as components of the NPC structure. Active, energy dependent nucleocytoplasmic transport of RNAs and proteins is mediated by a group of soluble receptor proteins, collectively termed karyopherins.
Gp210 has been suggested to be important for nuclear pore formation. Nevertheless, our analyses showed a limited expression pattern of gp210, with its mRNA and protein largely confined to epithelial cells in the mouse embryo. Furthermore, in several cell lines, gp210 was undetectable. The expression pattern of gp210 was not synchronised with some other nucleoporins, indicating NPC heterogeneity. Characterisation of the structure of the human gp210 gene, including its promoter region, gave insight about possible cell-type specific gene regulatory mechanisms.
Regulation of molecular traffic between the nucleus and the cytoplasm leads to transcriptional control. Cell specific configuration of the NPC structure, due to diffential expression of gp210, could be involved in this control. Gp210 could be of importance for the development of epithelial cell polarisation.
16
Asselman, Marino. "Hyaluronan biology and regulation in renal tubular epithelial cells and its role in kidney stone disease." [S.l.] : Rotterdam : [The Author] ; Erasmus University [Host], 2008. http://hdl.handle.net/1765/13147.
17
Hovater, Michael. "Underlying purinergic signaling important for monocilium-dependent signaling in ductal epithelia : implications for polycystic kidney disease." Thesis, Birmingham, Ala. : University of Alabama at Birmingham, 2006. http://www.mhsl.uab.edu/dt/2007m/hovater.pdf.
18
Abou, Samra Elias. "Elucidation of the Role of NKR‐P1: CLR Recognition Systems in Intestinal & Renal Epithelial Cell Homeostasis and Immunity." Thesis, Université d'Ottawa / University of Ottawa, 2017. http://hdl.handle.net/10393/35747.
Анотація:
Natural killer (NK) cells represent a crucial component of the innate immune system and are primarily regulated by the interactions of their activation and inhibitory receptors with ligands available on target cells. The genetically linked Ly49 and NKR-P1 family of receptors constitute two of the major regulatory receptor systems used by NK cells and have been shown to bind different ligands. Whereas the Ly49 receptors survey MHC-I ligands on target cells, the NKR-Pl receptor family members bind to various members of the C-type lectin-related (Clr) family. Interestingly, NKR-P1 and Clr haplotypes possess a stable genomic polymorphism across multiple mouse strains, suggesting that this inhibitory receptor:ligand relationship has an important role in the maintenance of host cellular cognate specificities. The NKR-P1 and Clr receptor-ligand pairs identified in mice include the NKR-P1B:Clr-b and the NKR-P1G:Clr-f interacting pairs. Previous RT-PCR and in situ RNA hybridization data generated by our laboratory determined that kidney tubular epithelium as well as the small and large intestinal epithelial cells specifically and highly expresses the Clr-f transcripts. Contrarily, the Clr-b transcripts were only detected on hematopoietic cells of various lymphoid organs and kidneys. Moreover, foregoing studies revealed that the loss of Clr-b following viral or chemical induced stress mediates NK cell killing of the target cell, suggesting a tissue-specific immune-surveillance mechanism in parallel with the global MHC-I-dependent missing-self model. However, the role of the NKR-P1B:Clr-b recognition-system have never been examined in the intestine. Additionally, the role of Clr-f in the kidney and intestines, where they are highly expressed, has not been investigated. For these reasons, I aimed in my thesis to provide a better understanding of the functional aspect of the NKR-P1B:Clr-b and NKR-P1G:Clr-f recognition systems in mediating gut mucosal and renal homeostasis, respectively.
First, in order to determine the role of NKR-P1B and Clrb receptor:ligand pair as a “missing-self” immunosurveillance system in the gut, I started by identifying the expression pattern of both the receptor and ligand on various intestinal cells. My results demonstrate that NK cells do not represent the major NKR-P1B-expressing cells in the gut lamina propria. Instead, ILC3 subsets constituted the predominant cell population expressing the receptor, whereas γδT cells composed a small fraction of NKR-P1B+ lymphocytes. In addition, the NKR-P1B expression on myeloid cells was exclusive to colon macrophages and DC subsets. Interestingly, the highest percentage of NKR-P1B+ immune cells was found in the gut, which suggests the dominant role of NKR-P1B in regulating immune functions at the level of intestinal mucosa. As expected, the expression of the NKR-P1B ligand, Clr-b, appeared on all innate immune cell types in the gut. Next, using oral infection models of Salmonela typhimurium and Citrobacter rodentium, I showed that NKR-P1B-deficient NK cells, ILC3 and γδ T cells are hyporesponsive compared to their WT counterparts. In particular, gut NKR-P1B-deficient NK cells and γδT cells secreted low levels of IFNγ cytokine while infected with S.typhimurium. Importantly, the decreased IFNγ secretion by NK and γδT cells was associated with an increased dissemination of the bacterium into the knockout spleens at day 5 post-infection. Likewise, I detected a significant decrease in IL-22 cytokine production by NKR-P1B-deficient ILC3 compared to their WT counterparts at both steady state and following C.rodentium infection.
Next, I address the potential role of Clr-f in the kidney. Renal tubular epithelial cells have been shown to express high levels of Clr-f transcripts. Epithelial cells constitute the major cellular component of kidney tubules and are well known to mediate metabolic waste excretion, reabsorption of essential molecules as well as other physiological functions, such as ions exchange and water retention. To determine the role of Clr-f in renal epithelial cells, I generated a Clr-f-deficient mouse with the help of two of my previous lab colleagues. Importantly, chemical analysis on urine and serum samples from knockout and WT littermates indicated that Clr-f-deficient kidneys display a decreased filtration capacity. In particular, higher creatinine levels were detected in the Clr-f deficient serum. In addition, Clr-f-deficient mice appeared to have a lower fractional excretion of sodium (FENa) in their urine filtrates in comparison to WT excreted urine. Blood pressure measurements on the same mice at 12 and 24 weeks of age revealed a hypotensive phenotype in the Clr-f-deficient mice. Furthermore, pathological assessment of Clr-f-deficient kidneys exhibited moderate and aggravated lesions of the tubular epithelium along with marked glomerular mesangiolysis. Lastly, flow cytometry analysis on isolated lymphocytes from Clr-f-deficient and WT mice demonstrated comparable immune infiltrates between the two mouse genotypes.
Altogether, our data shows that the absence of Clr-f results in the development of glomerular and tubular lesions in an immune-independent manner leading to an abnormal kidney function. Additionally, the disruption of NKR-P1B:Clr-b recognition system results in abnormal innate immune cell number and function in the mouse intestine. These novel findings sheds light on the important role of Clr-f in maintaining healthy kidney morphology and function, as well as the crucial role for NKR-P1B:Clr-b interactions in mediating intestinal homeostasis at steady and infected states.
19
Daniels, Nigel Allan. "Investigation of the role of novel SGK1 isoforms in regulation of sodium transport in kidney epithelial cells." Thesis, University of Newcastle Upon Tyne, 2010. http://hdl.handle.net/10443/936.
Анотація:
Serum/glucocorticoid regulated kinase 1 (SGK1) is a key component of the pathway that leads to activation of the epithelial sodium channel (ENaC) in the aldosterone-sensitive distal nephron (ASDN). Regulation of ENaC is a major determinant of renal Na+ absorption and overall body fluid homeostasis and blood pressure. Studies from our laboratory have shown that human skin cells express multiple SGK1 isoforms (A-F) that arise from alternative transcriptional start sites and RNA splicing at the SGK1 locus. The aim of this study was to investigate if SGK1 isoforms are also expressed in the ASDN and to assess their potential role in regulating Na+ transport. For these studies the mouse cortical collecting duct cell line mpkCCDcl4, was used as an in vitro model of the ASDN. Comparison of mouse Sgk1 expressed sequence tags (ESTs) with genomic DNA, identified four potential Sgk1 isoforms (Sgk1a-1d). Each isoform has a unique amino terminus of varying size, but otherwise an identical sequence. Using sequence specific primers, mRNA expression of all four isoforms was confirmed by RT-PCR from purified mpkCCDcl4 cell and mouse renal tissue RNA. mpkCCDcl4 cells exposed to aldosterone (Aldo) or Aldo plus insulin (Ins) showed a time-dependent increase in the endogenous expression levels of multiple Sgk1 bands within 1 hour of treatment. These Aldo and Aldo + Ins-induced endogenous Sgk1 bands co-migrated with overexpressed Sgk1 a-d isoform bands. Aldosterone also produced a significant increase in amiloride-sensitive (ENaCmediated) equivalent short circuit current within 2 hours of exposure, peaking after 4 hours. Insulin potentiated the Aldo response, but had no effect alone. Specific inhibitors showed that the hormonal response involved both PI3Kinase and mTOR-dependent and independent pathways. Immunofluorescence studies utilising cloned and tagged SGK1 isoforms in transfected HEK293T cells revealed cytoplasmic network-like staining for all SGK1 isoforms except for SGK1D, which produced plasma membrane staining. In mouse renal tissue, endogenous Sgk1d localised to the basolateral membrane of collecting duct epithelial cells. Furthermore co-immunoprecipitation of cloned human SGK1 and mouse Sgk1 proteins with the Aldo induced regulatory protein, glucocorticoid-induced leucine zipper protein 1 (GILZ1), showed isoform-specific interactions. Collectively, these results build upon our understanding of SGK1 gene expression, protein localisation and function in the ASDN.
20
Ohan, Nicholas. "An examination of small heat shock protein gene expression in Xenopus laevis embryos and A6 kidney epithelial cells." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1997. http://www.collectionscanada.ca/obj/s4/f2/dsk3/ftp04/nq21373.pdf.
21
Hambitzer, Martin. "Induction of interferon beta in human kidney epithelial cells by virulent and non-virulent strains of Escherichia coli." Thesis, Malmö högskola, Fakulteten för hälsa och samhälle (HS), 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:mau:diva-24963.
Анотація:
Urinvägsinfektioner (UVI) är ett vanligt hälsoproblem som drabbar miljontals människor. Den allvarligaste formen, akut pyelonefrit (APN) kan ge svåra komplikationer. Urinvägspatogena Escherichia coli (UPEC) som orsakar APN uttrycker P fimbrier som specifikt binder till glykosphingolipider på ytan av uroepitelceller. Det sätter igång en toll-like receptor 4 (TLR4) beroende men LPS-oberoende immunreaktion. Den roll som interferon beta (IFN-β) spelar vid bakterieinfektioner är inte helt klarlagd men studier som gjorts på IFN-β knockoutmöss visade på en ökad infektionsbenägenhet och svåra njursymptom vid infektion med UPEC. IFN-β uttrycket i uroepitelceller som svar på bakterieinfektion undersöktes. För att ta reda på om uttrycket är P fimbrieberoende infekterades humana A498 njurcarcinomceller med den P fimbrieförsedda pyelonefritstammen CFT073 eller den icke-virulenta asymtomatisk bakterieuristammen E. coli 83972 och inkuberades i 1,5 respektive 4 timmar. Som kontroll användes celler som enbart behandlats med PBS. Uttrycket av IFN-β analyserades med immunofluorescens (IF) och konfokalmikroskopi, samt med Western blot. Resultaten från konfokalmikroskopi visade att celler som exponerats för CFT073 under 4 timmar uttryckte mest IFN-β medan cellerna som utsatts för E. coli 83972 visade på ett omvänt förhållande. Western blot visade på högst uttryck i de E. coli 83972-behandlade cellerna. IFN-β uttrycktes i alla celler, inklusive kontrollcellerna, i någon utsträckning. Det kan betyda att IFN-β även induceras på någon alternativ väg och/eller att det uttrycks konstitutivt av njurepitelceller.Urinary tract infections (UTI) are a common health concern and affect millions of people. The most severe form of UTI, acute pyelonephritis (APN) is associated with serious complications. Uropathogenic Escherichia coli (UPEC) that cause APN express P fimbriae which specifically bind to glycosphingolipid molecules on the surface of urothelial cells. This triggers a toll-like receptor 4 (TLR4) mediated but LPS-independent innate immune response. The role of interferon beta (IFN-β) in bacterial infections is not well known but experiments with IFN-β knockout mice have shown an increased susceptibility and severe kidney pathology when infected with UPEC. IFN-β induction in urothelial cells in response to bacterial infection was investigated. To find out whether this response is P fimbriae dependent, A498 human kidney carcinoma epithelial cells were exposed to the P fimbriated CFT073 pyelonephritis strain or the non-virulent E. coli 83972 asymptomatic bacteriuria strain and incubated for 1.5 and 4 hours. For control, cells were treated with PBS alone. The IFN-β expression was analysed using immunofluorescence (IF) and confocal microscopy, and Western blot. Confocal microscopy results showed that the response to bacteria was both time- and dose-dependent. The highest IFN-β expression was detected in cells exposed to CFT073 for 4 hours, while cells exposed to E. coli 83972 showed an inverse relationship. Western blot analysis revealed that the highest expression was in the E. coli 83972 stimulated cells. IFN-β was expressed in all cells to some degree, including control cells. This could imply that IFN-β is induced by some other means and/or is constitutively expressed by kidney epithelial cells.
22
Polk, William Wyatt. "Role of protein kinase C zeta in lipopolysaccharide-mediated nuclear factor kappa B aactivation [i.e. activation] and aactivity [i.e. activity] in kidney epithelial cells /." Thesis, Connect to this title online; UW restricted, 2006. http://hdl.handle.net/1773/8463.
23
Sheremet, Andriy. "Bioinspired polyethersulfone-based hollow fiber membranes as the scaffolds in renal assist device for protein-bound toxins removal from blood." Master's thesis, Faculdade de Ciências e Tecnologia, 2014. http://hdl.handle.net/10362/13308.
Анотація:
Dissertation for obtaining the Master degree in Membrane EngineeringErasmus Mundus Master in Membrane Engineering
Using bioartificial kidney is the promising approach for removal of non-dializable, proteinbound uremic toxins, which are responsible for high mortality and morbidity in treating kidney failure related conditions. Additionaly, bioartificial kidney device could perform the physiological roles of the kidney such as metabolic replacement, endocrine function and immunomodulation. In the current work two commercial polyethersulfone-based membranes, Gambro HCO 1100 and Membrana MicroPES TF10 used in haemofiltration and plasma separation applications respectively were investigated. To provide adequate cytocompatibility of the membrane biomimetic, biomimetic double layer coating was developed. First, the membranes were coated with musselinspired synthetic polydopamine film, following with the coating of Collagen Type IV. Transport properties of the coated and native membranes were investigated. Increase in pure water permeability of the coated HCO 1100 membranes was observed. Membrane surface hydrophilization was assumed as the major factor responsible for the effect. Membrane permeabilities for bovine serum albumin and immunoglobulin G solutions were studied. Significant increase in protein rejection was observed for double coated HCO 1100 membranes with small or no effect of the double coated MicroPES TF10 membranes. Next, formation of confluent monolayers of the renal epithelial cells on the membrane scaffolds was studied. Cell seeding strategy was developed and two seeding conditions were tested. Specifically, the cells were allowed to adhere to the biomimetic membranes passively, and the negative pressure was applied to facilitate cell adhesion. After cultivation in semi-batch conditions the monolayer formation was examined. Confluent monolayers were observed for the conditions with passive cell adherence for the both membranes. Cell contacts formation and cell polarization were confirmed with the staining for ZO-1 protein. Applying the pressure to facilitate cell adhesion, on the contrary, resulted in the loss of cell ability to form functional monolayers.
EM3E Master is an Education Programme supported by the European Commission, the European Membrane Society (EMS), the European Membrane House (EMH), and a large international network of industrial companies, research centres and universities
24
Silva, Crysthiane Saveriano Rubião. "Apoptose precoce, proliferação celular sincrônica tardia e perfil de expressão de proteínas ao complexo esclerose tuberosa e às doenças renais policísticas durante tubulogênese in vitro." Universidade de São Paulo, 2013. http://www.teses.usp.br/teses/disponiveis/5/5160/tde-01082013-145925/.
Анотація:
O complexo esclerose tuberosa (CET) e as doenças renais policísticas autossômica dominante (DRPAD) e autossômica recessiva (DRPAR) são doenças monogênicas associadas a cistogênese renal. Os produtos dos genes mutados nessas enfermidades, respectivamente tuberina e hamartina para CET, policistina-1 (PC1) e policistina-2 para DRPAD, e poliductina/fibrocistina para DRPAR, modulam proliferação, diferenciação, apoptose, crescimento e/ou migração celular. Neste estudo empregamos um sistema tridimensional de cultura de células IMCD para caracterizar os perfis de expressão dessas proteínas durante a tubulogênese. Usando uma matriz de colágeno tipo I/Matrigel e fator de crescimento de hepatócito (HGF), a formação de estruturas alongadas se iniciou dois dias após o plaqueamento in vitro (2 DIV), ao passo que o desenvolvimento de lúmen ocorreu entre 10-14 DIV. A marcação para caspase-3 ativa foi mais intensa nas fases iniciais da tubulogênese, enquanto a marcação para Ki-67 foi uniformemente pronunciada em estágios mais tardios. A tuberina e a hamartina apresentaram expressão citoplasmática e co-localização acentuada em 6 e 12 DIV. A PC1 apresentou maior expressão nas porções ramificadas dos túbulos que nas não ramificadas no 12 DIV, um padrão não verificado para a PC2. Estas proteínas exibiram expressão citoplasmática, assim como expressão ocasional e pontual na membrana plasmática. PD1 também apresentou expressão citoplasmática. Nossos dados sugerem que a apoptose e a ciclagem celular sincrônica durante a tubulogênese in vitro são mais acentuadas, respectivamente, em fases mais precoces e mais tardias da formação tubular. Nossos achados demonstram, além disso, que as proteínas relacionadas ao CET e às DRPs são expressas in vitro durante a tubulogênese, apoiando um papel importante para a interação tuberina-hamartina na formação tubular, e são consistentes com o padrão de expressão diferencial da PC1 observado durante a nefrogêneseTuberous sclerosis complex (TSC) and autosomal dominant and recessive polycystic kidney diseases (ADPKD and ARPKD) are monogenic diseases associated with renal cystogenesis. The products of the genes mutated in these disorders, respectively tuberin and hamartin for TSC, and polycystin-1 (PC1), polycystin-2 (PC2) and polyductin/fibrocystin (PD1) for PKD, modulate cell proliferation, differentiation, apoptosis, growth and/or migration. We have employed an IMCD tridimensional cell culture system to characterize their expression profiles along tubulogenesis. Using a type I collagen/Matrigel matrix and hepatocyte growth factor (HGF), the formation of elongated structures initiated 2 days after in vitro plating (2 DIV) while lumen developed between 10-14 DIV. Active caspase-3 labeling was more intense in initial phases of tubulogenesis while Ki-67 staining was uniformly pronounced in later stages. Tuberin and hamartin showed cytoplasmic expression and marked co- localization at 6 and 12 DIV. PC1 displayed higher expression in branching than non- branching portions of the tubules at 12 DIV, a pattern not verified for PC2. These proteins presented cytoplasmic and occasional, punctate membrane expression. PD1 also showed cytoplasmic expression. Our data suggest that apoptosis and synchronous cell cycling during in vitro tubulogenesis are more remarkable, respectively, in early and later steps of tubule formation. In addition, our findings demonstrate that the TSC and PKD proteins are expressed in vitro during tubulogenesis, supporting an important role for tuberin-hamartin interaction in tubular formation, and are consistent with the differential PC1 expression pattern observed during nephrogenesis
25
Kozlov, Vladimir. "Étude du rôle des gènes SoxC au cours du développement du rein de souris." Thesis, Université Côte d'Azur, 2020. http://www.theses.fr/2020COAZ6010.
Анотація:
Les anomalies congénitales des reins et des voies urinaires (CAKUT) sont un groupe de malformations fréquemment trouvées chez les patients humains qui résultent de défauts dans le programme de développement de ces organes. Sox11 fait partie de la famille des facteurs de transcription SoxC, qui joue un rôle important dans le développement de divers organes chez les vertébrés. Les souris Sox11-/- meurent juste après la naissance et présentent une grande variété de CAKUT. Alors que les reins duplex sont une anomalie courante du développement rénal, le mécanisme moléculaire qui amène les mutants Sox11 à développer cette malformation reste incertain. Dans ce projet, j'ai analysé le phénotype rénal des souris Sox11-/- et montré que les reins duplex sont causés par l'expansion du mésenchyme métanéphrique (MM). D'autres expériences ont été réalisées pour déterminer si l'origine de cette expansion résidait dans une apoptose insuffisante ou dans l'absence de migration des cellules du MM. Une analyse tridimensionnelle par microscopie confocale des cellules apoptotiques a révélé que chez les embryons Sox11-/- le MM subissait une apoptose dans la région d'intérêt similaire à celles des embryons de type sauvage. Un test in vitro de cicatrisation des plaies a démontré que les gènes SoxC sont nécessaires pour la migration des cellules du MM. La délétion de ces gènes à un stade tardif du développement conduit à une hypoplasie rénale et à une diminution du nombre de néphrons due à l'arrêt de la néphrogenèse. Pour étudier le rôle des gènes SoxC au cours de la formation des reins, j'ai établi une culture de cellules progénitrices rénales permettant l’invalidation de ces gènes en présence de 4OH-Tamoxifène. L'analyse de l'expression génique a confirmé la suppression efficace des gènes SoxC, tout en maintenant l'identité des cellules progénitrices du néphron. Après induction de la délétion, ces cellules sont incapables de s'épithélialiser, du fait d’une activité accrue de la voie Wnt/β-caténine. Afin de valider ces résultats et d’identifier les cibles en aval des gènes SoxC, j'ai effectué une analyse ARN-seq d’organoïdes rénaux subissant une transition mésenchymateuse-épithéliale. Les données acquises par ces expériences révèlent que de nombreuses voies de signalisation sont affectées. En particulier, l’insuffisance du gène Ezh2 codant pour une protéine du groupe Polycomb, pourrait être à l’origine de changements transcriptomiques généralisés empêchant les cellules progénitrices déficientes en SoxC de se différencier. L’ensemble de ces données démontre l'importance des gènes de la famille SoxC dans le développement précoce et tardif du rein, dont l'analyse moléculaire conduira à des orientations possibles pour les recherches et les traitements futursCongenital Anomalies of Kidneys and Urinary Tract (CAKUT) are a group of birth defects that arise from defects in the developmental program of organ development and are frequently found in human patients. Sox11 is a member of SoxC family of transcription factors, which plays an important role in the development of various organs in vertebrates. Sox11 knockout mice die soon after birth and display a wide variety of CAKUT, including duplex kidneys and nephrogenesis defects. While duplex kidneys are a common renal development anomaly, the molecular mechanism causing Sox11 mutants to develop duplex kidneys remains unclear. In this project, I analyzed the renal phenotype of Sox11 knockout mice and discovered that the duplex kidneys are caused by the expansion of metanephric mesenchyme (MM). Further experiments were performed to determine whether the origin of this expansion lies in insufficient apoptosis of MM cells or lack of MM cell migration. Three-dimensional confocal microscopy analysis with Lysotracker Red staining of apoptotic cells revealed that MM undergoes apoptosis in the region of interest in wildtype embryos, however the apoptosis seems to persist in Sox11-/- embryos as well. However, in vitro scratch wound assay provided evidence that SoxC genes are necessary for migration of MM cells. In the late kidney development, SoxC deletion is leading to renal hypoplasia and reduced nephron number due to cessation of nephrogenesis. To study the role of SoxC in nephrogenesis, I set up an in vitro renal progenitor cell culture which allows for the timed deletion of SoxC genes by a 4OH-tamoxifen induced knockout. Gene expression analysis confirmed efficient deletion of SoxC genes, while maintaining nephron progenitor cell identity. After deletion, renal progenitor cells were found to be unable to epithelialize, linked with increased activity of Wnt/β-catenin pathway. To validate these findings and discover the downstream targets of SoxC genes, I developed a system for an RNA-seq analysis of renal organoids undergoing mesenchymal-to-epithelial transition. Data acquired in these experiments reveal numerous regulatory pathways affected in SoxC-knockout renal progenitor cells, with the deficiency of Polycomb group gene Ezh2 as a likely origin of widespread transcription changes leading to the inability of SoxC-deficient progenitor cells to differentiate. Taken together these data demonstrate the importance of Sox11 and other SoxC genes in early and late kidney development, with molecular analysis providing plausible directions for future research and interventions
26
Schnatwinkel, Carsten. "Characterisation of Novel Rab5 Effector Proteins in the Endocytic Pathway." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2004. http://nbn-resolving.de/urn:nbn:de:swb:14-1106824900192-45576.
Анотація:
Endocytosis, a process of plasma membrane invaginations, is a fundamental cellular mechanism, ensuring uptake of nutrients, enhanced communication between cells, protective functions against invasive pathogens and remodelling of the plasma membrane composition. In turn, endocytic mechanisms are exploited by pathogens to enter their host cells. Endocytosis comprises multiple forms of which our molecular understanding has mostly advanced with respect to clathrin-mediated endocytosis and phagocytosis. Studies on the small GTPase Rab5 have provided important insights into the molecular mechanism of endocytosis and transport in the early stages of the endocytic pathways. Rab5 is a key regulator of clathrin-mediated endocytosis, but in addition, localises to several distinct endocytic carriers including phagosomes and pinocytic vesicles. On early endosomes, Rab5 coordinates within a spatially restricted domain enriched in phosphatidylinositol-3 phosphate PI(3)P a complex network of effectors, including PI3-Kinase (PI3-K), the FYVE-finger proteins EEA1 and Rabenosyn-5 that functionally cooperate in membrane transport. Moreover, Rab5 regulates endocytosis from the apical and basolateral plasma membrane in polarised epithelial cells. During my PhD thesis, I investigated the molecular mechanisms of endocytosis both in polarised and non-polarised cells. I obtained new insights into the molecular mechanisms of endocytosis and their coordination through the functional characterization of a novel Rab5 effector, termed Rabankyrin-5. I could demonstrated that Rabankyrin-5 is a novel PI(3)P-binding Rab5 effector that localises to early endosomes and stimulates their fusion activity in vitro. The latter activity depends on the oligomerisation of Rabankyrin-5 on the endosomal membrane via the N-terminal BTB/POZ domain. In addition to early endosomes, however, Rabankyrin-5 localises to large vacuolar structures that correspond to macropinosomes in epithelial cells and fibroblasts. Overexpression of Rabankyrin-5 increases the number of macropinosomes and stimulates fluid phase uptake whereas its downregulation through RNA interference inhibits these processes. In polarised epithelial cells, the function of Rabankyrin-5 is primarily restricted to the apical membrane. It localises to large pinocytic structures underneath the apical surface of kidney proximal tubule cells and its overexpression in polarised MDCK cells specifically stimulates apical but not basolateral, non-clathrin mediated pinocytosis. In demonstrating a regulatory role in endosome fusion and (macro)-pinocytosis, my studies suggest that Rab5 regulates and coordinates different endocytic mechanisms through its effector Rabankyrin-5. Furthermore, the active role in apical pinocytosis in epithelial cells suggests an important function of Rabankyrin-5 in the physiology of polarised cells. The results obtained in this thesis are central not only for our understanding of the basic principles underlying the regulation of multiple endocytic mechanisms. They are also relevant for the biomedical field, since actin-dependent (macro)-pinocytosis is an important mechanism for the physiology of cells and organisms and is upregulated under certain pathological conditions (e.g. cancer).
27
Nicholson, Benjamin. "The regulation of high affinity glutamate transport a bovine renal epithelial cell line." Thesis, University of Bristol, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.336919.
28
Takayama, Akira. "Transport of cyclosporin A in kidney epithelial cell line (LLC-PK[1])." Kyoto University, 1995. http://hdl.handle.net/2433/160728.
Анотація:
本文データは平成22年度国立国会図書館の学位論文(博士)のデジタル化実施により作成された画像ファイルを基にpdf変換したものであるKyoto University (京都大学)
0048
新制・論文博士
博士(医学)
乙第8919号
論医博第1514号
新制||医||613(附属図書館)
UT51-95-P410
(主査)教授 藤田 潤, 教授 吉田 修, 教授 乾 賢一
学位規則第4条第2項該当
29
Parry, Robin Geoffrey. "Cytokines in minimal change nephropathy." Thesis, University of Bristol, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.341511.
30
Shukla, D. H. "Manipulation of the VHL/HIF pathway in mouse kidney epithelia and pancreatic β-cells". Thesis, University College London (University of London), 2011. http://discovery.ucl.ac.uk/1306810/.
Анотація:
Biallelic inactivation of the von Hippel-Lindau tumour suppressor gene (VHL) underlies both, sporadic clear cell renal cell carcinoma (ccRCC) that arises as part of an inherited multi-cancer VHL syndrome. Pancreatic tumours also occur in VHL disease and are thought to be of neuroendocrine origin. The most extensively studied role of VHL is hypoxia-inducible-factor (HIF) regulation. The HIF pathway is known operates in all cell types examined to date and plays a central role in many physiological responses to oxygen concentration. This thesis investigates the role of the Vhlh tumour suppressor protein in the kidney and pancreatic β-cells of mice. Cell-specific inactivation of Vhlh in mice was obtained using the Cre/loxP system. Analysis from mice constitutively expressing an active form of the HIF-2α transgene (KsptmHIF2α) was also performed. Furthermore, I have also used a doxycycline controlled Cre transgene system to allow temporal control of Vhlh ablation in the kidney. The main results arising from this thesis are: Vhlh inactivation in the distal tubular segment of the nephron led to an increase in both HIF isoforms (HIF-1α and HIF-2α) accompanied with upregulation of downstream gene expression. Constitutive expression of HIF-2α in the distal tubular segment upregulated downstream gene expression but did not lead to cyst or tumour formation; interestingly, inactivating another tumour suppressor gene, fumarate hydratase (Fh1) in the mouse kidney, resulted in cyst formation accompanied by HIF activation. Taken together, these findings suggest that HIF is not solely responsible for cyst formation in the Fh1 knockout mice, as cysts were not seen with the Vhlh knockout mice. The other major finding arising from this thesis is that, intact Vhlh is necessary for glucosestimulated- insulin secretion. I show that this is mediated via HIF transactivation and involves a glucose transporter switch, from glucose transporter–2 (GLUT-2) to glucose transporter–1 (GLUT-1). In addition, I have found that the expression of microRNA is altered in a murine pancreatic β-cell line, MIN6 in response to hypoxia. Therefore, the work arising from this thesis provides significant insights into understanding how the VHL/HIF pathway can be used to identify early morphological alterations, leading to a more aggressive phenotype or tumourigenesis. This pathway also plays a crucial role in glucose homeostasis, and interference in this pathway causes defects in β-cell function and insulin secretion.
31
Ryan, Sean P. "Autosomal Recessive Polycystic Kidney Disease Epithelial Cell Model Reveals Multiple Basolateral EGF Receptor Sorting Pathways." Case Western Reserve University School of Graduate Studies / OhioLINK, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=case1274887553.
32
Huynh, Carl. "The cytoprotective role of Ras signaling in glomerular epithelial cell injury /." Thesis, McGill University, 2007. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=112639.
Анотація:
In experimental membranous nephropathy, complement C5b-9-induced glomerular epithelial cell (GEC) injury leads to breakdown of glomerular peimselectivity and proteinuria. This study addresses mechanisms that limit complement-mediated injury, focusing on Ras. Complement-mediated injury was attenuated in cultured GEC expressing a constitutively active form of Ras (V12Ras), compared with Neo (control) GEC. V12Ras GEC showed constitutive activation of phosphatidylinositol 3-kinase and extracellular signal-regulated kinase pathways, but inhibition of these pathways did not reverse the protective effect of Ras. V12Ras GEC showed smaller and rounder morphology, decreased F- to G-actin ratio, decreased activity of the Rho GTPase, Rac, and decreased Src activity. In V12Ras GEC, disruption or stabilization of the F-actin cytoskeleton reversed the protective effect of V12Ras on complement-mediated injury. Thus, the protective effect of V12Ras may be dependent on remodeling of the actin cytoskeleton. Furthermore, the reduction of Src activity due to Ras activation may alter the equilibrium in activities of Rho GTPases, a family of proteins known regulate the actin cytoskeleton. Activation of Ras signaling is a novel pathway to consider in developing strategies for cytoprotection in complement-mediated injury.
33
Li, Moying [Verfasser], and Hans-Joachim [Akademischer Betreuer] Anders. "Mdm2 prevents spontaneous tubular epithelial cell death and acute kidney injury / Moying Li ; Betreuer: Hans-Joachim Anders." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2019. http://d-nb.info/1186629444/34.
34
Naillat, F. (Florence). "Roles of Wnt4/5a in germ cell differentiation and gonad development & ErbB4 in polarity of kidney epithelium." Doctoral thesis, Oulun yliopisto, 2011. http://urn.fi/urn:isbn:9789514295751.
Анотація:
Abstract The embryonic urogenital system generates the metanephric kidneys, the gonads and the adrenal glands, and its development is based on sequential and reciprocal cell and tissue interactions. The mechanisms which regulate urogenital ontogeny are still poorly understood. In this thesis, the roles of Wnt-4 and ErbB4 functions in gonad and kidney development were analysed by using in vivo functional genomic technologies. Wnt-4 is crucial in female development since its absence leads to a partial female to male sex reversal. We found that Wnt-4 mediated the interactions between the somatic and the germ cells and played a role in meiosis which is regulated in part by the secreted signal retinoic acid (RA). Expression of certain meiosis-controlling genes (Stra8, Spo11) was inhibited in the Wnt-4 deficient germ cells, while certain pluripotency genes (Oct4, Fgf9, Sox2 and Dnmt3l) were activated similarly as in the wild-type male gonad. In addition to this, we noted that a gene encoding for a Cyp26b1 enzyme, which degrades RA in the embryonic testis, was ectopically expressed in the Wnt-4 deficient ovary. Microarray analysis was used to identify candidate Wnt-4 target genes by using the Wnt-4 knock-out mouse. Of these genes, Runx-1 may represent a novel signalling target to mediate Wnt-4 activity in the control female development The role of receptor-tyrosine kinase ErbB4 in kidney development was studied by using both in vivo gain and loss of function approaches. In the gain-of-function situation, we found that certain markers for the epithelial tubules and collecting ducts lost their polarized expression pattern. At the same time, the orientation of the cells in the kidney tubules was deregulated and an increase in cell proliferation was noticed. We suggest that the observed defects gave rise to an increase in the tubule diameter and to cyst formation in the kidney cortex. In the loss-of-function mouse, the lack of ErbB4 expression led to a similar phenotype as with the gain of function, and the renal functions of the mutant adult kidneys were compromised. In conclusion, the results point to specific roles for Wnt-4 and ErbB4 in the control of urogenital development. Wnt-4 appears to be crucial in sustaining proper female somatic cell and germ cell differentiation, and maintenance of gonad development during and after the sex determination event, while ErbB4 activity is critical for the regulation of tubular growth in embryonic kidney developmentTiivistelmä Sekä nisäkkään jälkimunuainen, lisämunuainen että sukurauhanen kehittyvät alkion urogenitaalialueen järjestelmästä ja solu- ja kudosvuorovaikutukset ohjaavat elinkehitysprosessia. Tapahtuman molekyylitason mekanismit ovat kuitenkin huonosti tunnettuja. Tässä väitöskirjatyössä tutkittiin Wnt-4 signaalin tehtäviä sukurauhasen ja ErbB4- proteiinin munuaisen kehityksessä. Wnt-4 signaali on keskeinen naisen sukupuolisuuden kehityksessä, koska signaalin puutos aiheuttaa alkion sukupuolen osittaisen kääntymisen naaraasta koiraaksi. Tarkastelimme aluksi sitä, välittääkö Wnt-4 itusolujen ja sukurauhasen somaattisten solujen vuorovaikutuksia ohjaten itusolujen meioosia, jota mm. A-vitamiini säätelee. Havaitsimme, että Wnt-4 geeni puuttuessa tietyt meioosia säätelevät geenit kuten Stra8 ja Spo11 olivat heikentyneet, kun taas solujen monikykyisyyteen liittyvät geenit kuten Oct4, Fgf9, Sox2 ja Dnmt3l aktivoituivat vastaavalla tavalla kuin havaitaan normaalisti koirasalkion kivesaiheessa. Tämän lisäksi havaitsimme, että Cyp26b1-geeni, joka johtaa A-vitamiinin hajoamiseen alkiossa ja estää normaalisti meioosin koirasalkion kivesaiheessa oli aktivoitunut munuaisrauhasaiheessa, jolta puuttuu Wnt-4 aktiivisuus. Tuloksemme osoittavat, että Wnt-4 säätelee osaltaan naarasalkion itusolujen meioosia. Tarkastelimme myös mikrosirututkimusten avulla niitä geenejä, joita Wnt-4 säätelee sukuelinaiheessa. Identifioimme useissa Wnt ja β-catenin signaalireittiin liittyvissä geeneissa muutoksia. Muuntuneet geenit voivat olla Wnt-4 signaalireitin kohdegeenejä. Näistä Runx-1 saattaa olla keskeinen Wnt signaalitien kohdegeeni, joka säätelee merkittävällä tavalla naaraan munarauhasen kehitystä. Väitöskirjan toisessa osassa tarkastelimme ErbB4-reseptorityrosiinikinaasin tehtäviä munuaisen kehityksen säätelyssä. ErbB4-geenin tehtäviä tutkittiin käyttäen hyväksi siirtogeenisiä malliorganismeja, joissa ErbB4-geenin määrä oli joko koholla tai ajastetusti inaktivoitu. ErbB4- geenin kokeellinen yliaktiivisuus muutti spesifisti tekijöitä, jotka säätelevät osaltaan jälkimunuaisen epiteeliputkien solujen orientaatiota ja solun jakautumista. Solujen orientaatiomuutoksen yhteydessä myös solujen jakautuminen häiriintyi. Oletuksemme on, että nämä epiteelikudoksessa tapahtuneet muutokset ovat syy, miksi kohotettu ErbB4-aktiviteetti muuttaa epiteeliputkien paksuutta ja pituutta erityisesti munuaisen pintakerroksissa. Havaitsimme myös, että ErbB4-geenin ajastettu poistaminen munuaisen epiteelikudoksessa johti hyvin samankaltaisiin, mutta vastakkaisiin muutoksiin kuin ErbB4-aktiviteetin kohottaminen. Muutokset johtivat myös muutoksiin munuaisen toiminnassa. Yhteenvetona toteamme, että näillä Wnt-4 ja ErbB4 solusignallointiin liittyvillä molekyyleillä on keskeinen tehtävä alkion munarauhasen ja munuaisen aiheen kehityksen säätelyssä. Wnt-4 ohjaa sekä itusolujen että somaattisten solujen erilaistumista ja samalla sukupuolen määräytymistä ja jatkokehitystä, kun taas ErbB4-signallointireseptorin tehtävä on avainasemassa munuaisen epiteeliputken kasvun säätelyssä
35
Rondeau, Eric. "Les activateurs du plasminogene du rein humain : identification, localisation, facteurs de secretion." Paris 6, 1987. http://www.theses.fr/1987PA066607.
Анотація:
Les glomerules isoles du cortex renal chez l'homme, contiennent des activateurs du plasminogene. L'activateur tissulaire du plasminogene (t-pa) represente 80% de l'activite profibrinolytique et l'eurokinase 20% de cette activite. Les cellules epitheliales glomerulaires en culture contiennent l'urokinase. L'activite profibrinolytique des surnageant glomerulaires augmente lorsque les glomerules sont incubes en presence du calcium et a ph alcalin. Les acides gras polyinsatures (acide arachidonique, acide eicosopentaenoique ou acide eicosatrienoique) augmentent cette activite alors que les acides gras satures n'ont pas d'effet
36
Murugan, S. (Subramanian). "Control of nephrogenesis by Wnt4 signaling:mechanisms of gene regulation and targeting of specific lineage cells by tissue engineering tools." Doctoral thesis, Oulun yliopisto, 2012. http://urn.fi/urn:isbn:9789526200323.
Анотація:
Abstract Wnt4, a member of the Wnt family of secreted factors, is essential for kidney organogenesis since the kidney fails to develop in its absence. Besides the kidney, Wnt4 signaling is involved in the control of development of several other organs such as the gonads, adrenal glands and pituitary gland. In the context of the embryonic kidney, Wnt4 signaling induces mesenchymal to epithelial transition of the progenitor cells in the metanephric mesenchyme, an early step in nephrogenesis. Wnt4 signaling may also be relevant in the development of a childhood kidney tumor, the Wilms’ tumor, that involves the function of Wilms’ tumor suppressor protein 1 (WT1). Wilms’ tumor is thought to arise from the early metanephric mesenchymal cells of the embryonic kidney, but the detailed mechanisms are not known. The main aim of this project was to study the mechanisms that regulate expression of the Wnt4 gene by using immortalized embryonic kidney mesenchyme-derived mK4 cells as a model. The Wnt4 gene expression was also analyzed in vivo in the frog embryonic pronephros. Through the use of reporter assays and a two-hybrid screen, Sox11, a member of the SoxC family of transcription factors, was identified as a synergistic protein that interacts with WT1. Immunoprecipitation studies provided further evidence that Sox11 and WT1 may physically interact with each other in the developing embryonic kidney. Indeed, Sox11 and WT1 may regulate the Wnt4 gene expression in vivo since the morpholino-based knock-down of either WT1 or Sox11 led to notable downregulation of the Wnt4 gene expression in the frog embryonic pronephros. The other general aim of this thesis was to develop novel tissue targeting and therapy tools to the cell lineages regulated by the Wnt4 signals, including the podocytes. For this purpose, we utilized mice carrying a floxed expression cassette for the avidin-LDL receptor fusion protein, Lodavin, in the constitutively active Rosa-26 locus. Three Cre driver mice, including the Wnt4-Cre knock-in line, were used to activate Lodavin expression in the respective cells of the embryonic kidney. Moreover, we generated a podocyte injury model by expressing the human receptor for diphtheria toxin specifically in the podocytes. This was achieved by crossing mice containing a floxed expression cassette for this receptor in the Rosa-26 locus with those expressing the Cre recombinase under the nephrin promoter. Administration of diphtheria toxin led initially to podocyte damage only, followed by a progression to glomerular sclerosis. As a summary, Sox11 and WT1 serve as synergistic transcription factors that may regulate expression of the Wnt4 gene in vivo. The transgenic mouse models generated and used provide the basis to generate acute and chronic kidney disease models and the potential to purify the respective cells for developing cell-based therapy avenues for the kidney. Moreover, the Lodavin-based approaches may enable targeted delivery of biotinylated small compounds, proteins, viruses or even cells and novel means for in vivo imaging and functional studiesTiivistelmä Wnt-4 kuuluu signaloivien proteiinien Wnt-perheeseen ja sen toiminta on välttämätöntä munuaisen kehityksessä. Ilman Wnt-4 proteiinia munuainen ei kehity. Munuaisen lisäksi Wnt4-signalointi on mukana useiden muiden elinten, kuten sukurauhasten, lisämunuaisen ja aivolisäkkeen säätelyssä. Alkion munuaisessa Wnt4-signalointi saa aikaan mesenkymaalisen kantasolukon epitelisoitumisen, edustaen näin ollen nefronin kehityksen varhaisia vaiheita. Wnt4-signaloinnilla on myös merkittävä asema lapsuusiän munuaiskasvaimen, niin kutsutun Wilmsin kasvaimen kehittymisessä. Tämän tyyppisessä kasvaimessa keskeisenä on Wilmsin tuumoriproteiinin WT1:n toiminta, mutta myös Wnt4:n toiminnalla voi olla merkitystä. Wilmsin kasvaimen arvellaan saavan alkunsa varhaisista sikiöaikaisista jälkimunuaisen soluista, mutta yksityiskohtaisia mekanismeja ei vielä tunneta. Tämän projektin tarkoituksena oli tutkia Wnt4-geenin ilmentymistä sääteleviä mekanismeja käyttäen mallina mK4-soluja eli alkion munuaisesta saatuja, immortalisoituja soluja. Wnt4-geenin ilmentymistä analysoitiin myös in vivo sammakon alkion alkumunuaisessa. Tuplahybridi-analysoinnin avulla tunnistettiin transkriptiotekijäperhe SoxC:n jäsen Sox11 samantoimiseksi proteiiniksi transkriptiotekijä WT1:n kanssa Wnt4-geenin ilmentymisen säätelyssä. Immunopresipitaatiotutkimukset tukivat ajatusta, että Sox11 ja WT1 voisivat olla fyysisessä vuorovaikutuksessa säädellessään nefroninmuodostuksen alullepanossa ratkaisevan Wnt4-geenin ilmentymistä. Sox11 ja WT1 voivat mahdollisesti säädellä Wnt4-geenin ilmentymistä myös in vivo, sillä morfoliineihin perustuvissa kokeissa sekä WT1:n että Sox11:n hiljennys laski Wnt4-geenin ilmentymistasoa sammakon alkumunuaisessa. Tämän väitöstutkimuksen toinen yleinen tavoite oli kehittää uusia kudoskohdennus- ja terapiakeinoja Wnt4-signaloinnin säätelemille solulinjoille, kuten podosyyteille. Tätä tarkoitusta varten kloonattiin siirtogeeninen hiiri, jossa floksattu avidiini-LDL -reseptorifuusioproteiini, Lodavin, kohdennettiin jatkuvasti aktiiviseen Rosa-26 -lokukseen. Kolmea eri Cre-hiirilinjaa käytettiin aktivoimaan Lodavinin ilmentyminen kussakin tietyssä alkion munuaisen solupopulaatiossa. Yksi näistä Cre-linjoista oli Wnt4-Cre. Jotta kyettäisiin vahingoittamaan samoja soluja, jotka ilmentävät Lodavinia, hyödynnettiin difteriamyrkyn ihmisen reseptoria (iDTR). IDTR:n ilmentäminen tietyissä hiiren soluissa tekee ne alttiiksi tappavalle difteriamyrkylle. IDTR-perusteisen munuaisvauriomallin kehittämiseksi käytettiin floksattua iDTR-hiirimallia, ja geenin ilmentyminen aktivoitiin Wnt4-indusoiduissa munuaissolulinjoissa, erityisesti podosyyteissä Nephrin Cre -välitteisesti. NephrinCre;R26RiDTR hiiriä altistettiin difteriamyrkylle ja niiden munuaiskerästen muutoksia seurattiin. Tutkimukset antavat viitteitä siitä, että R26R-floksatut iDTR-hiiret toimivat hyvänä mallina kehitettäessä sekä akuutteja että kroonisia munuaistautimalleja. Yhteenvetona voidaan todeta, että Sox-11 ja WT-1 ovat samantoimisia transkriptiotekijöitä, jotka voivat säädellä Wnt4-geenin ilmentymistä in vivo. Tutkimuksessa kehitetyt ja käytetyt siirtogeeniset hiirimallit tarjoavat perustan kehittää sekä akuutteja että kroonisia munuaistautimalleja. Samalla ne mahdollistavat kulloistenkin solujen eristämisen uusien soluperusteisten hoitomenetelmien kehittelemiseksi. Lisäksi Lodavin-perusteiset lähestymistavat voivat mahdollistaa biotinyloitujen pienten yhdisteiden, proteiinien, virusten tai jopa solujen kuljetuksen kohdennetusti sekä avata uusia mahdollisuuksia in vivo -kuvantamiselle ja toiminnallisille tutkimuksille
37
Toutain, Hervé. "Développement et caractérisation de modèles expérimentaux pour l'étude ex-vivo et in-vitro de la cellule tubulaire proximale de rein de lapin." Rouen, 1989. http://www.theses.fr/1989ROUES036.
Анотація:
Une suspension de cellules tubulaires proximales de rein de lapin est isolée par une nouvelle méthode qui ne fait pas intervenir d'enzymes protéolytiques. Ces cellules, après une caractérisation biochimique, morphologique, et métabolique, sont séparées en deux populations hautement purifiées par une technique d'électrophorèse ou flux libre en veine liquide. Présentation d'un modèle de culture primaire de cellules tubulaires proximales de rein de lapin
38
Friedlander, Gérard. "Etude des facteurs qui modulent l'effet des hormones sur des cellules epitheliales d'origine renale." Paris 7, 1987. http://www.theses.fr/1987PA077204.
39
Yang, An-Hang. "Reactive oxygen metabolism in cultured kidney epithelial cells." 1987. http://catalog.hathitrust.org/api/volumes/oclc/16310902.html.
40
Ren, Shuyu. "The regulation of Nephrin expression in kidney epithelial cells and during inflammatory kidney diseases /." 2005. http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&doc_number=014191117&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA.
41
Dai, Long-jun. "Control of intracelluar Ca²⁺ in epithelial cells of the kidney thick ascending limb." Thesis, 1995. http://hdl.handle.net/2429/7260.
Анотація:
The cortical thick ascending limb of Henle’s loop (cTAL) plays a fundamental role in
salt reabsorption and concentration-dilution processes within the nephron. This study was
performed to characterize the role of intracellular free Ca²⁺ ([Ca²⁺]i) in hormone-mediated
signal transduction and to describe the function of Na⁺/Ca²⁺ exchange in control of [Ca²⁺]i
in isolated cTAL cells from porcine kidney. Parathyroid hormone, arginine vasopressin, and
atrial natriuretic peptide transiently increased [Ca²⁺]i, in a dose-dependent manner. The
increment in [Ca²⁺]i induced by these hormones was by intracellular release and entry
through plasma membrane Ca²⁺ channels. These hormone-induced Ca²⁺ transients were
modulated by cAMP, cGMP, and PKC activation. In order for intracellular Ca²⁺ to play
a role in signal transduction mechanisms it is necessary to have regulated processes which
maintain [Ca²⁺]i at submicromolar levels. We evaluated the functional role of Na⁺/Ca²⁺
exchange in cTAL cells. cTAL cells treated with ouabain had basal [Ca²⁺]i 86±2 nM.
Removal of external Na⁺ or voltage depolarization with KC1 resulted in rapid and
reversible maximal elevation of [Ca²⁺]i 1023 ±72 nM (n=28), which was dependent on the
presence of external Ca²⁺ and elevated [Na⁺]i. The activity of Na⁺/ Ca²⁺ exchange was
modulated by protein phosphorylation as calmodulin inhibition decreased and phosphatase
inhibition increased the apparent exchange activity. The presence of a Na⁺/ Ca²⁺ exchanger
was confirmed with northern hybridization techniques. A gene transcript which encodes
a portion of the intracellular ioop of the renal Na⁺/ Ca²⁺ exchanger was amplified from
cortical tissue and cTAL cells by polymerase chain reaction (PCR) using primers flanking
the alternative splicing site. Southern hybridization and DNA sequencing demonstrated
the isoform contained exons B and D characteristic of one isoform (NACA3) of the renal Na⁺/ Ca²⁺ exchanger. The results provide both functional and molecular evidence for a
N2aICa exchanger in cTAL cells of the porcine kidney. It is likely that Na⁺/ Ca²⁺
exchange plays an important role in [Ca²⁺]i control and thus hormonal regulation of
electrolyte reabsorption within the cTAL cells.
42
Music, Ena. "Stress-induced accumulation of heme oxygenase-1 in Xenopus laevis A6 kidney epithelial cells." Thesis, 2014. http://hdl.handle.net/10012/8403.
Анотація:
Abstract
Previous studies have examined stress-induced heme oxygenase-1 (HO-1) expression primarily in mammalian systems. The present study examines, for the first time in amphibians, the effect of heat shock, sodium arsenite, cadmium chloride, and the proteasomal inhibitor MG132 on HO-1 accumulation in Xenopus laevis A6 kidney epithelial cells. Western blot analysis revealed that exposure of A6 cells to a range of heat shock temperatures (30-35 °C), which induced HSP30 accumulation, did not induce HO-1 accumulation. In contrast, cells treated with sodium arsenite (5-50 μM), cadmium chloride (50-200 μM) or MG132 (5-30 μM) exhibited a dose- and time-dependent accumulation of HO-1. Additionally, immunocytochemical analysis revealed that HO-1 and HSP30 accumulation occurred in a granular pattern primarily in the cytoplasm in cells treated with sodium arsenite, cadmium chloride, or MG132. In cells recovering from sodium arsenite or cadmium chloride treatment, HO-1 and HSP30 accumulation initially increased to a maximum at 12 h and 24 h recovery, respectively, followed by a 50% reduction at 48 h. This initial increase in the relative levels of stress proteins was likely the result of new synthesis as it was inhibited by cycloheximide. In comparison, cells recovering from MG132 treatment displayed reduced but prolonged accumulation of HO-1 and HSP30. Interestingly, cells treated with low concentrations (10 μM) of sodium arsenite or MG132 but not cadmium chloride in combination with a mild 30 °C heat shock had enhanced accumulation of HO-1 and HSP30 accumulation compared to either of the stressors individually. This study has shown for the first time in amphibians that HO-1 accumulation is induced in response to metals and proteasomal inhibitors, suggesting that it may play a role in mediating the cellular stress response in X. laevis.
43
Payne, Emily Harman. "Polyploidy and Mitotic Cell Death are Two Distinct HIV-1 Vpr-Driven Outcomes in Renal Tubule Epithelial Cells." Diss., 2016. http://hdl.handle.net/10161/12190.
Анотація:
Given the emerging epidemic of renal disease in HIV+ patients and the fact that HIV DNA and RNA persist in the kidneys of HIV+ patients despite therapy, it is necessary to understand the role of direct HIV-1 infection of the kidney. HIV-associated kidney disease pathogenesis is attributed in large part to viral proteins. Expression of Vpr in renal tubule epithelial cells (RTECs) induces G2 arrest, apoptosis and polyploidy. The ability of a subset of cells to overcome the G2/M block and progress to polyploidy is not well understood. Polyploidy frequently associates with a bypass of cell death and disease pathogenesis. Given the ability of the kidney to serve as a unique compartment for HIV-1 infection, and the observed occurrence of polyploid cells in HIV+ renal cells, it is critical to understand the mechanisms and consequences of Vpr-induced polyploidy.
Here I determined effects of HIV-1 Vpr expression in renal cells using highly efficient transduction with VSV.G pseudotyped lentiviral vectors expressing Vpr in the HK2 human tubule epithelial cell line. Using FACS, fluorescence microscopy, and live cell imaging I show that G2 escape immediately precedes a critical junction between two distinct outcomes in Vpr+ RTECs: mitotic cell death and polyploidy. Vpr+ cells that evade aberrant mitosis and become polyploid have a substantially higher survival rate than those that undergo complete mitosis, and this survival correlates with enrichment for polyploidy in cell culture over time. Further, I identify a novel role for ATM kinase in promoting G2 arrest escape and polyploidy in this context. In summary, my work identifies ATM-dependent override of Vpr-mediated G2/M arrest as a critical determinant of cell fate Vpr+ RTECs. Further, our work highlights how a poorly understood HIV mechanism, ploidy increase, may offer insight into key processes of reservoir establishment and disease pathogenesis in HIV+ kidneys.
Dissertation
44
Lin, Pei-ying, and 林佩瑩. "Effect of zingerone on the anti-oxidation and anti-inflammatory in rat kidney epithelial cells." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/62233197664593538772.
Анотація:
碩士大仁科技大學
食品科技研究所
98
Ginger is a common spice used in popular cuisine and has been widely applied in research of diseases due to its rich ingredients. The purpose of this research is to probe into the antioxidation activity and anti-inflammatory effect of zingerone (ZO) of NRK 52E cells (Normal rat kidney epithelial cells). The experiment design covers two parts: the first test is “In vitro” in which ZO of different concentrations (i.e. 1, 5, 7.5, 10 mM) are used to analyze DPPH activity, SOD, FIC and reducing power. The results show that ZO demonstrates good effects in removing DPPH activity and reducing power. Moreover, the ZO 10 mM has the highest removal rate of 19% on SOD activity; however, the ZO demonstrates no FIC ability. The second test is the “Ex-vivo” in which t-BHP 2 mM is used to induce NRK 52E oxidative injuries and ZO of different concentrations (i.e. 1, 5, 7.5, 10 mM) are added and cultured at 37°C to evaluate the antioxidation ability after 15, 30, and 60 minutes. With antioxidation analysis indicators including MTT assay and TBARs and GSH, the results show that ZO causes no effect on the survival rate of cells. As ZO increases in concentration, the production of TBARs is significantly reduced and GSH expression is enhanced. In addition, the IL-1β 5 ng is used to induce NRK 52E inflammation and ZO of different concentrations (i.e. 1, 5, 7.5, 10 mM) are added and cultured at 37°C to evaluate the anti-inflammatory ability after 20 hours. Measured analysis of the survival rate, NO, iNOS, COX-2, PGE2, IκB, p-IκB and NF-κB. The results showed that ZO 1, 5, 7.5, 10 (mM) compared with the control group, the survival rate is no change, in IL-1β + ZO this group 4 there are cells to reduce the generation of NO, iNOS, PGE2 and COX-2 performance (p < 0.05). On the other hand, p-IκB and NF-κB performance has also been decline. The study, ZO inhibits inflammation caused by IL-1β induced NRK 52E to achieve anti-inflammatory effect. At the same time, it is able to inhibit oxidative injuries caused by t-BHP induced NRK 52E thus protecting kidney cells.
45
Woolfson, Jessica Pearl. "Examination of Cadmium-Induced Heat Shock Protein Gene Expression in Xenopus laevis A6 Kidney Epithelial Cells." Thesis, 2008. http://hdl.handle.net/10012/3723.
Анотація:
Cadmium is a highly toxic chemical and has been classified by the International Agency for Research on Cancer as a human carcinogen. Cadmium is abundant in the environment, at specific work places, and in food and water. Toxicological responses to cadmium exposure include respiratory diseases, neurological disorders and kidney damage. The present study examined the effects of cadmium on heat shock protein (HSP) accumulation in Xenopus laevis A6 kidney epithelial cells. HSPs are molecular chaperones involved in protein folding and translocation. In response to environmental stress these proteins bind to unfolded protein and inhibit their aggregation. Stress-inducible hsp gene transcription is mediated by the heat shock promoter element (HSE), which interacts with heat shock transcription factor (HSF). In the present study, hsp30 and hsp70 mRNA and protein were induced by heat shock, as determined by northern and western blot analysis. Exposure of A6 cells to cadmium chloride also induced the expression of hsp genes. For example, northern and western blot analysis revealed that exposure of A6 cells to cadmium chloride induced the accumulation of hsp30 and hsp70 mRNA and their respective proteins. Western blot analysis also revealed that A6 cells recovering from a cadmium chloride treatment retained relatively high levels of HSP30 and HSP70 protein accumulation over 24 h after the removal of the stress. Treatments combining a mild heat shock and cadmium chloride resulted in a synergistic increase in hsp30 and hsp70 gene expression at mRNA and protein levels. Further experiments in which two stressors were combined revealed that synergistic effects occurred with varying cadmium concentrations and different temperatures. Immunocytochemistry and confocal microscopy were used to confirm the results attained from western blot analysis. Further, this technique allowed the determination of intracellular localization of HSP30 in A6 cells and the examination of cellular morphology and cytoskeletal structure during cadmium chloride treatments. A 2 h heat shock at 33°C resulted in the accumulation of HSP30 in the cytoplasm, whereas a 2 h heat shock at 35°C resulted in some HSP30 accumulation in the peripheral region of the nucleus. This is in contrast to cells treated with cadmium chloride, where HSP30 accumulation was restricted to the cytoplasm. A 14 h 50 μM cadmium chloride treatment resulted in the accumulation of HSP30 in approximately 10% of cells. The proportion of cells displaying HSP30 accumulation increased to 80% and 95% in cells treated with 100 μM and 200 μM, respectively. HSP30 accumulation frequently occurred in large granular structures. High concentrations of cadmium chloride resulted in cell membrane ruffling at areas of cell-cell contact, as well as actin disorganization. This study characterized the pattern of hsp gene expression, accumulation and localization under various cadmium chloride conditions. These results suggest that hsp30 and hsp70 gene expression can be used as potential biomolecular markers for cadmium exposure.
46
Khan, Saad. "Examination of curcumin-induced heat shock protein gene expression in Xenopus laevis A6 kidney epithelial cells." Thesis, 2010. http://hdl.handle.net/10012/5338.
Анотація:
The heat shock response is a cellular homeostatic mechanism that is activated in response to stressful stimuli (e.g. heat shock, heavy metals, disease states etc.), which causes an increase in unfolded protein, which triggers the expression of heat shock protein (hsp) genes. HSPs are molecular chaperones that assist in protein synthesis, folding and degradation and prevent stress-induced protein aggregation. Since stressor-induced tissue damage is associated with different disease states, indirect evidence suggested that HSP inducers may be therapeutically beneficial for certain diseases. Curcumin, a phenolic compound found in the Indian spice, Curcuma longa (Turmeric), was shown to have anti-inflammatory, anti-tumor and anti-amyloid properties. In the present study, it was determined that curcumin inhibited the ubiquitin-proteasome system (UPS) activity and induced the accumulation of HSPs in the frog model system, Xenopus laevis. Treatment of A6 kidney epithelial cells with curcumin enhanced ubiquitinated protein levels and inhibited chymotrypsin-like activity. Furthermore, HSP30 and HSP70 accumulation was observed in cells exposed to 10 - 50 μM curcumin for 24 h in a concentration-dependent manner with maximal levels of HSP30 and HSP70 in cells treated with 30 μM curcumin. Time-dependent increases in HSP30 and HSP70 accumulation were also observed in cells treated with 30 μM curcumin for 2 to 24 h. The accumulation of HSP30 and HSP70 in cells recovering from curcumin exposure increased up to 24 h after treatment. The simultaneous treatment of A6 cells with 10 μM curcumin and mild heat shock (30 ºC) for 6 h resulted in an enhanced accumulation of HSP30 and HSP70, which was greater than with each stressor alone. This pattern of combined stressor-induced HSP30 and HSP70 accumulation increased from 2 to 6 h, after which it decreased from 10 to 24 h. The activation of HSF1 may be involved in curcumin-induced hsp gene expression in A6 cells since KNK437, a heat shock factor-1 inhibitor, inhibited the accumulation of HSP30 and HSP70. Immunocytochemical analysis employing the use of laser scanning confocal microscopy (LSCM) revealed that curcumin-induced HSP30 was detectable primarily in the cytoplasm in a punctate pattern with minimal detrimental effects on the actin cytoskeleton. Elevation of the incubation temperature from 22 to 30 °C greatly enhanced the curcumin-induced cytoplasmic accumulation of HSP30 in a granular pattern. Lastly, curcumin treatment also conferred a state of thermotolerance in A6 cells such that they were able to maintain proper actin cytoskeleton in subsequent thermal challenges. This phenomenon was controlled at the transcriptional level since pretreatment of cells with KNK437, repressed HSP30 accumulation and cytoprotection. These findings are of importance given the interest in identifying agents that can upregulate HSP levels with minimal effects on cell structure or function as a therapeutic treatment of certain protein folding diseases.
47
Khan, Saad. "Analysis of heat shock protein 30 gene expression and function in Xenopus laevis A6 kidney epithelial cells." Thesis, 2014. http://hdl.handle.net/10012/8398.
Анотація:
Heat shock proteins (HSPs) are molecular chaperones that assist in protein synthesis, folding and degradation and prevent stress-induced protein aggregation. The present study examined the pattern of accumulation of HSP30 and HSP70 in cells recovering from heat shock as well as the effect of proteasome inhibition on cytoplasmic/nuclear and endoplasmic reticulum (ER) molecular chaperone accumulation, large multimeric HSP30 complexes, stress granule and aggresome formation in Xenopus laevis A6 kidney epithelial cells. Initial immunoblot analysis revealed the presence of elevated levels of HSP30 after 72 h of recovery. However, the relative levels of HSP70 declined to near control levels after 24 h. The relative levels of both hsp30 and hsp70 mRNA were reduced to low levels after 24 h of recovery from heat shock. Pretreatment of cells with cycloheximide, a translational inhibitor, produced a rapid decline in HSP70 but not HSP30. The cycloheximide-associated decline of HSP70 was blocked by the proteasomal inhibitor, MG132, but had little effect on the relative level of HSP30. Also, treatment of cells with the phosphorylation inhibitor, SB203580, in addition to cycloheximide treatment enhanced the stability of HSP30 compared to cycloheximide alone. Immunocytochemical studies detected the presence of HSP30 accumulation in a granular pattern in the cytoplasm of recovering cells and its association with aggresome-like structures, which was enhanced in the presence of SB203580. To verify if proteasome inhibition in A6 cells induced the formation of similar HSP30 granules, immunoblot and immunocytochemical analyses were performed. MG132, celastrol and withaferin A enhanced ubiquitinated proteins, inhibited chymotrypsin-like activity of the proteasome and induced the accumulation of cytoplasmic/nuclear HSPs, HSP30 and HSP70 as well as ER chaperones, BiP and GRP94 and heme oxygenase-1. Northern blot experiments determined that proteasome inhibitors induced an accumulation in hsp30, hsp70 and bip mRNA but not eIF1α. The final part of this study demonstrated that treatment of A6 cells with proteasome inhibitors or sodium arsenite or cadmium chloride induced HSP30 multimeric complex formation primarily in the cytoplasm. Moreover, these stressors also induced the formation of RNA stress granules, pre-stalled translational complexes, which were detected via TIA1 and polyA binding protein (PABP), which are known stress granule markers. These stress granules, however, did not co-localize with large HSP30 multimeric complexes. In comparison, proteasome inhibition or treatment with sodium arsenite or cadmium chloride also induced the formation of aggresome-like structures, which are proteinaceous inclusion bodies formed as a result of an abundance of aggregated protein. Aggresome formation was identified by monitoring the presence of vimentin and γ-tubulin, both of which are cytoskeletal proteins and serve as markers of aggresome detection. Aggresome formation, which was also verified using the ProteoStat assay, co-localized with large HSP30 multimeric complexes. Co-immunoprecipitation experiments revealed that HSP30 associated with γ-tubulin and β-actin in cells treated with proteasome inhibitors or sodium arsenite or cadmium chloride suggesting a possible role in aggresome formation. In conclusion, this study has shown that the relative levels of heat shock-induced HSP30 persist during recovery in contrast to HSP70. While HSP70 is degraded by the ubiquitin-proteasome system, it is likely that the presence of HSP30 multimeric complexes that are known to associate with unfolded protein as well as its association with aggresome-like structures may delay its degradation. Finally, proteasome inhibition, sodium arsenite and cadmium chloride treatment of A6 cells induced cytoplasmic/nuclear and ER chaperones as well as resulting in the formation stress granules and aggresome-like structures which associated with large HSP30 multimeric complexes.
48
Bellin, Gloria. "Heparanase regulates M1 macrophage polarization and the crosstalk between macrophages and tubular epithelial cells after kidney ischemia/reperfusion injury." Doctoral thesis, 2018. http://hdl.handle.net/11562/985324.
Анотація:
l’HPSE è un elemento chiave coinvolto nei complessi processi biologici attivati dal danno da I/R, regolando la attivazione/polarizzazione dei macrofagi e il crosstalk tra queste cellule infiammatorie e l’epitelio tubulare renale.
49
Tai, Wei-Chun, and 戴偉竣. "The differential effect of hexavalent and trivalent chromium on the growth of human kidney epithelial cells and their respective biological significance." Thesis, 2015. http://ndltd.ncl.edu.tw/handle/43158578602201785994.
Анотація:
碩士國立中興大學
生命科學院碩士在職專班
103
Following advanced industrialization of the new century, as well as the modernization of family utensils and consumables, many heavy metals, which consist of beneficial effects, such as heat-, corrosion and rust-resistance, are gradually forced into everybody’s everyday life. In particular, stainless pipes, water faucets, stainless cooking wares, stainless drinking machines, blenders, and kitchen knives that contain metallic chromium are becoming indispensable. In fact, chromium, as essential microelements in human bodies, plays significant roles in the metabolism of ingested sugars and fats. In natural foods, trivalent chromium [Cr(III)] has been frequently detected in beer yeasts, honey, dry cheeses, eggs, apple peels, bananas, wheat flour, potatoes, meat products and livestock entrails, suggesting the biological significance of Cr(III) in the Nature. However, the naturally most prevalent hexavalent chromium [Cr(VI)] is toxic to the human bodies, which could directly damage human lungs, gastrointestinal tracts and kidneys as well as increase carcinogenesis risks of those organs. Inhaled chromium dusts could markedly elevated lung cancer incidences. Even traced amounts of chromium in the cigarette or from electric welding could be accumulated first in the lungs before absorbing through pulmonary alveoli into the circulation system, and distributed to the important organs, including kidneys. Although following chromium damage, kidney progenitor cells, which are closely associated with glomerular and tubular repairs, have been identified, the biological effect of Cr(VI) on these cells have not been well studied. Our lab had recently characterized these cells and their biological responses post-chromium insults. In addition, the biological function of Cr(III) in the formation of DNA/RNA adducts that could strengthen DNA/RNA structural stabilization to prevent genetic mutations as well as carcinogenesis intrigues us to pursue their difference from DNA/RNA adducts formed by polycyclic aromatic hydrocarbons (PAH). Therefore, the aims of this study are to investigate the differential effect of hexavalent and trivalent chromium on the growth of human kidney epithelial cells and their respective biological significance. Our results showed that Cr(VI) could increase expressions of epithelial-to mesenchymal transition and stem cell markers of human kidney epithelial HK-2 cells. Such reactions could be spontaneous responses of human kidney epithelial cells. The biological and medical applications of these findings to the regeneration of kidney epithelium and repair of kidney functions could be tremendous. Moreover, these results could be applied to the skin maintenance and restoration. As we already know that several kinds of ceramide have been used for cosmetic moisturization, and these ceramides, in particular, the glycosylated ceramides, are nature products of the skin, which are secreted by the epidermis to protect underlying growing basal layers via affecting ion channels. Nonetheless, exposure to ceramide could activate ATG6 to provoke cell autophagy, which corresponded well with our observations that showed Cr(III) could induce massive autophagy as well. Cr(III)-induced autophagy, however, did not directly lead to cell apoptosis or cell death. Our result further support the previous findings in application of Cr(III) in prevention of type II diabetes. Such theories could be applied to the youthful skin maintenance. Organic trivalent chromium could be complimentary to the use of ceramide, or Cr(III) could replace ceramide usage directly in the future to induce functional cosmetic autophagy as well as to maintain organelle activities and metabolism of proteins and lipids.
50
Brunt, Jara. "Examination of sodium arsenite- and cadmium chloride-induced HSP accumulation and inhibition of proteasome activity in Xenopus laevis A6 kidney epithelial cells." Thesis, 2011. http://hdl.handle.net/10012/6283.
Анотація:
Sodium arsenite (NA) and cadmium chloride (CdCl2) are two relatively abundant environmental toxicants that have numerous detrimental effects on living organisms. At the cellular level, NA and CdCl2 produce reactive oxygen species which cause protein damage. Recent studies, in mammalian systems, have suggested that NA and CdCl2 can inhibit the ubiquitin-proteasome system (UPS). The UPS is the major degradation route for the elimination of damaged protein. This process involves two successive steps: the addition of ubiquitin to damaged protein and subsequent degradation by the 26S proteasome. Previously our laboratory determined that inhibition of the UPS can induce the accumulation of heat shock protein (HSPs) in Xenopus laevis A6 cells. HSPs are molecular chaperones, which bind to unfolded proteins to prevent aggregation and assist in protein refolding. The goal of this study was to examine the effect of NA and CdCl2 on the UPS of Xenopus laevis cells and to investigate any possible association between HSP accumulation and proteasomal activity. In the present study, treatment of A6 cells with NA or CdCl2 caused an increase in HSP30 and HSP70 accumulation, as well as in protein ubiquitination. In fact, treatment with 30 μM NA or 200 μM CdCl2 resulted in a 1.7- and 2-fold increase, respectively, in the relative levels of ubiquitinated protein compared to control. Examination of the relative levels of ubiquitinated protein over time revealed significant increases in cells treated up to at least 24 h after exposure of A6 cells with 30 μM NA or 200 μM CdCl2. To further examine the effect of NA and CdCl2 on proteasome activity, a cell-based assay measuring proteasomal chymotrypsin (CT)-like activity was employed. Treatment with 20 or 30 μM NA caused a 40 % decrease in the relative levels of chymotrypsin (CT)-like activity in A6 cells compared to control cells. The CT-like activity of A6 cells treated with 100 or 200 μM CdCl2 decreased by 40 % and 75 %, respectively, compared to control. The increase in ubiquitinated protein and a decrease in CT-like activity suggest that NA and CdCl2 can inhibit the UPS in A6 cells. In order to examine any possible association between HSP accumulation and proteasome activity an inhibitor of HSP accumulation, KNK437, was employed. In A6 cells pretreated with KNK437 followed by NA or CdCl2 exposure a decrease in the relative levels of HSP30 and HSP70 and ubiquitinated protein was noted compared to cells treated with NA alone. However, the CT-like activity of cells pretreated with KNK437 prior to NA or CdCl2 showed no significant difference compared to cells treated with NA or CdCl2 without pretreatment. The findings of this study suggest that NA and CdCl2 inhibit proteasomal activity in A6 cells and that there is a possible association between HSP accumulation and the mechanism by which damaged proteins are ubiquitinated.