Identification oftargetproteins for endocrine disrupters in the testis

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The use of in vitro cell culture models has been of central importance in the development of cellular and molecular biology of Organs and tissues. Transgenic mouse and rat harboring temperature-sensitive simian virus 40 large T-antigen gene are useful for establishing cell lines from organs and tissues that have proved difficult to culture in vitro. Many conditionally immortalized cell lines such as mouse gastric epithelial, testicular Sertoli and Leydig cells were generated from the transgenic animals. DNA microarray technology has broad applications, and its power is directed toward the study of global gene expression. Using established cell lines and DNA microarrays, we identified many genes that were up-and down-regulated in the process of the cell differentiation or cell death.We examined the time course of changes in gene expression in detail using cDNA microarray analysis of mouse Sertoli TTE3 cells treated with bisphenol A(BPA). A subtoxic dose of BPA(200μM) transiently increased intracellular Ca^<2+> and time-dependently induced an increase in mRNA level of 78-kDa glucose-regulated protein, indicating that BPA induces endoplasmic reticulum stress. Of the 865 genes analyzed, 31 genes showed increased levels of expression. TaqMan analysis confirmed that the mRNA levels of chop, fra-2, c-myc, and ornithine decarboxylase were increased, and showed that chop-10 is the most sensitive gene. The expression level of chop protein and cell injury induced by BPA were significantly reduced in stable TTE3 cells overexpressing full-length chop antisense RNA. We conclude that chop plays a key role in Sertoli cell injury induced by BPA.