ABSTRACT
BACKGROUND: Cocaine- and amphetamine-regulated transcript (CART), discovered initially by via differential display RT-PCR analysis of brains of rats administered cocaine, is expressed mainly in central nervous system or neuronal origin cells, and is involved in a wide range of behaviors, such as regulation of food intake, energy homeostasis, and reproduction. The hens egg-laying rate mainly depends on the developmental status of follicles, expression of CART have not been identified from hen follicles, the regulatory mechanisms of CART biological activities are still unknown. The objective of this study was to characterize the mRNA expression of CART in hen follicular granulosa cells and determine CART peptide localization and regulatory role during follicular development. METHODS: Small white follicles (1-2 mm in diameter) were treated for RNA isolation; Small white follicles (1-2 mm in diameter) and large white follicles (4-6 mm in diameter) were treated for immunohistochemical localization and large white follicles (4-6 mm in diameter), small yellow follicles (6-8 mm in diameter), large yellow follicles (9-12 mm in diameter), mature follicles (F5, F4, F3, F2, F1, >12 mm in diameter) were treated for RNA isolation and Real time PCR. RESULTS: The results showed that full length of the CDS of hen CART was 336 bp encoding a 111 amino acid polypeptide. In the hen ovary, CART peptide was primarily localized to the theca layer, but not all, the oocyte and granulosa layer, with diffused, weaker staining than relative to the theca cell layer. Further, amount of CART mRNA was more (P < 0.05) in granulosa cells of 6-8 mm follicles compared with that in granulosa cells of other follicles. However, CART mRNA amount was greater in theca cells of 4-6 mm follicles relative to follicles of other sizes (P < 0.05). CONCLUSIONS: Results suggest that CART could play a potential role in developmental regulation of chicken follicles.
Subject(s)
Animals , Female , Ovarian Follicle/metabolism , Nerve Tissue Proteins/metabolism , Immunohistochemistry , Cells, Cultured , Chickens , DNA, Complementary/biosynthesis , Reverse Transcriptase Polymerase Chain Reaction , Gene Expression Profiling , Nerve Tissue Proteins/geneticsABSTRACT
Objective:To investigate the localization and expression pattern of nuclear receptor NR4A1 protein in follicle and corpus luteum cells at different estrous stages in cycling female SD rat. Methods:The SD experimental rats were divided into four ovarian stages based on the vaginal lavages,from which the ovaries were obtained in every cycling stage.The localization and expression pattern of NR4A1 in rat ovaries were characterized by immunohistochemistry. Results:NR4A1 was mainly expressed in the cytosol of ovarian theca cells and lutein cells.Expression level of NR4A1 in theca cell of mature follicle was significantly higher than that in the growing follicle(P
ABSTRACT
To understand the role of IGF2 gene in development of human ovary, IGF2 expression was detected by monoclonal antibody for IGF2 to its producted protein with immunohistochemical technique on human ovarian tissues. The results was as follows. IGF2 was highly expressed in ovum of mature follicle, IGF2 expression, however, was not high in granulosa and the cells. IGF2 was not highly expressed in ovum of primary follicle. Highly expressed IGF2 was found on corpus luteum and no expression of IGF2 was found in stroma and epithelial cells. These results suggest that IGF2 is important role in ovulation and in production of progesterone. Abnormal IGF2 expression may be concerned to carcinogenesis of ovarian tumor because most of all tumor from ovary is originated from epithelium.