Isolation of novel candidate genes involved in bovine adipocyte differentiation and marbling using a clonal bovine intramuscular preadinocyte (BIP) cell line

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To order to isolate candidate genes involved in bovine adipocyte differentiation, we have constructed a subtraction library from a clonal bovine intramuscular preadipocyte (BIP) cell line using the suppression subtractive hybridization method. We have isolated a set of subtracted cDNA fragments whose respective mRNA levels are up regulated during the adipogenic differentiation of BIP cells, and cloned a full-length cDNA from differentiated-BIP-lambda ZAP II cDNA library. The clone 4-3 was isolated and was highly homologous to the sequence of human, mouse and rat collagen prolyl 4 hydroxylase alpha III subunit (C-P4HIII). C-P4HIII is known to refold collagens and collagen like proteins, and as necessary for folding collagen triple helices structure. From the expression analysis of C-P4HIII mRNA and protein during adipogenesis in BIP cells, C-P4HIII expression was elevated both mRNA and protein. It is known that extracellular matrix (ECM) of cell surface has been restructured during adipogenesis. The ECM was decomposed by adipogenic induction, and up-regulated expression of components, included collagens for the adipocyte. These results suggest that C-P4HIII may be associated with adipocyte differentiation and adipose formation in cattle and is a potentially useful marker for adipogenesis.