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

Chaki, Sulalita
http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1282134876

Abstract Details

2010, Master of Science (MS), Ohio University, Biological Sciences (Arts and Sciences).
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.
Karen Coschigano, PhD (Advisor)
Kenneth Goodrum, PhD (Committee Member)
Ramiro Malgor, PhD (Committee Member)
Darlene Berryman, PhD (Committee Member)
87 p.
Biology
growth hormone; GH; mesangial cell; STAT5; MAPK

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Citations

  • Chaki, S. (2010). Characterization of Mesangial Cell Lines Established from Nontransgenic (NT) and Growth Hormone Receptor Knockout (GKO) Mice [Master's thesis, Ohio University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1282134876

    APA Style (7th edition)

  • Chaki, Sulalita. Characterization of Mesangial Cell Lines Established from Nontransgenic (NT) and Growth Hormone Receptor Knockout (GKO) Mice. 2010. Ohio University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1282134876.

    MLA Style (8th edition)

  • Chaki, Sulalita. "Characterization of Mesangial Cell Lines Established from Nontransgenic (NT) and Growth Hormone Receptor Knockout (GKO) Mice." Master's thesis, Ohio University, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1282134876

    Chicago Manual of Style (17th edition)

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