The mammary alveolar cell-T (MAC-T) cell line is able to uniformly differentiate and secrete casein proteins in response to dexamethasone, insulin and prolactin and is extensively used to study bovine mammary epithelial cell function. Growth hormone (GH) has been shown to increase milk protein synthesis both in vivo and in mammary cell models, and induce cytoskeletal rearrangement in 3T3 fibroblast cell line and a Chinese hamster ovary (CHO) cell line. Few studies have focused on identifying the mechanisms involved in differentiated MAC-T cells’ response to GH. We tested the hypothesis that MAC-T cells would respond directly to GH and that the response would include alterations in milk protein gene expression, leading to a more appropriate model for mammary cell function than treatment with dexamethasone, insulin and prolactin alone. To identify mechanisms that are involved in MAC-T cells’ response to GH, global protein was assessed through two-dimensional gel electrophoresis and differentially expressed proteins were identified through mass spectrometry. Differentiated cells expressed GH receptor mRNA, and addition of GH to the differentiation medium increased production of α-S1 casein and α-lactalbumin mRNA. Proteins that were differentially expressed are related to metabolism, the cytoskeleton, protein folding, RNA and DNA processing, detoxifying and calcium metabolism. These results indicate that GH is an important factor in inducing a lactogenic phenotype in the MAC-T cell line, and supports GHs involvement in differentiation, while altering cell metabolism in preparation for synthesis and secretion of milk components.
| Identifer | oai:union.ndltd.org:CALPOLY/oai:digitalcommons.calpoly.edu:theses-1362 |
| Date | 01 June 2010 |
| Creators | Johnson, Tasha Lynn |
| Publisher | DigitalCommons@CalPoly |
| Source Sets | California Polytechnic State University |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Master's Theses |
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