Characterization of Mesangial Cell Lines Established from Nontransgenic (NT) and Growth Hormone Receptor Knockout (GKO) Mice

Characterization of Mesangial Cell Lines Established from Nontransgenic (NT) and Growth Hormone Receptor Knockout (GKO) Mice

Bibliographic Details
Main Author: Chaki, Sulalita
Language:English
Published: Ohio University / OhioLINK 2010
Subjects:
Biology
growth hormone
GH
mesangial cell
STAT5
MAPK
Online Access:http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1282134876
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spelling ndltd-OhioLink-oai-etd.ohiolink.edu-ohiou12821348762021-08-03T05:46:45Z Characterization of Mesangial Cell Lines Established from Nontransgenic (NT) and Growth Hormone Receptor Knockout (GKO) Mice Chaki, Sulalita Biology growth hormone GH mesangial cell STAT5 MAPK Growth hormone (GH) is a peptide hormone that is secreted from the anterior pituitary under the regulation of different proteins, hormones, amino acids and nutrients.GH and its signaling pathways regulate cellular growth, development and metabolism.Involvement of GH in damaging the kidney, especially in diabetes, is a matter of concern as diabetic nephropathy has been regarded as an increasing health threat. Establishment of mesangial cell lines could provide an effective in vitro model to investigate a connection between individual/multiple downstream signaling pathway/s of GH signaling and damage of kidney mesangium and thus was the topic of this thesis. The primary step of establishing this in vitro model was to characterize the presence or absence of functional GH signaling in mesangial cell lines established from the nontransgenic (NT) and growth hormone receptor gene-disrupted or knockout (GKO) mice. Glomeruli from the kidneys of NT and GKO mice were isolated and used to establish two mesangial cell lines. Specific morphology, selective media and immunohistochemistry confirmed the identity of the cells as mesangial cells. Next, expressed GHR transcripts were characterized in the mesangial cell lines by performing RT/PCR and gel electrophoresis. Expression of an intact GHR transcript in NT mesangial cells and a stable but mutated GHR transcript in GKO cells was demonstrated. Finally, the presence or absence of GH signaling in NT and GKO mesangial cell lines was tested by assaying the induction of RNA transcripts or phosphorylated proteins of downstream signaling pathways in response to exogenous GH stimulation using real-time RT/PCR, western blotting analysis or ELISA. The only response observed was GH stimulation of iNOS and STAT5b mRNA expression in both the NT and GKO mesangial cell cultures, suggesting that mesangial cells from the NT mice might be useful for studying the GH-stimulated expression of specific GH responsive genes but the cells from the GKO mice may not provide the expected contrast of lack of GH signaling. It will be important to further evaluate GH signaling and responses in both the NT and GKO mesangial cells to firmly establish the role of GH in mesangial cells. 2010-09-22 English text Ohio University / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1282134876 http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1282134876 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection NDLTD
language English
sources NDLTD
topic Biology
growth hormone
GH
mesangial cell
STAT5
MAPK
spellingShingle Biology
growth hormone
GH
mesangial cell
STAT5
MAPK
Chaki, Sulalita
Characterization of Mesangial Cell Lines Established from Nontransgenic (NT) and Growth Hormone Receptor Knockout (GKO) Mice
author Chaki, Sulalita
author_facet Chaki, Sulalita
author_sort Chaki, Sulalita
title Characterization of Mesangial Cell Lines Established from Nontransgenic (NT) and Growth Hormone Receptor Knockout (GKO) Mice
title_short Characterization of Mesangial Cell Lines Established from Nontransgenic (NT) and Growth Hormone Receptor Knockout (GKO) Mice
title_full Characterization of Mesangial Cell Lines Established from Nontransgenic (NT) and Growth Hormone Receptor Knockout (GKO) Mice
title_fullStr Characterization of Mesangial Cell Lines Established from Nontransgenic (NT) and Growth Hormone Receptor Knockout (GKO) Mice
title_full_unstemmed Characterization of Mesangial Cell Lines Established from Nontransgenic (NT) and Growth Hormone Receptor Knockout (GKO) Mice
title_sort characterization of mesangial cell lines established from nontransgenic (nt) and growth hormone receptor knockout (gko) mice
publisher Ohio University / OhioLINK
publishDate 2010
url http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1282134876
work_keys_str_mv AT chakisulalita characterizationofmesangialcelllinesestablishedfromnontransgenicntandgrowthhormonereceptorknockoutgkomice
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