Expression Of CART Gene And Protein Locating In Ovary Of Hen

Cocaine-and Amphetamine-Regulated Transcript (CART) has caused great concern of researchers, and widely research on drug addiction, diet and obesity, endocrine and so on since it was found. Kobayashi found CART had negative regulatory role on bovine ovary and caused follicular atresia. Taking into accont the affect of CART on breeding, Hen was selected as the experimental animal in the tets, got sequences of CART by using gene cloning and compared homology with other species; identified the protein expression by using immunohistochemistry technique in hen ovary; using Real-time PCR to research expression patterns of CART in granulose cells and theca cells of hen ovary, aimed to investigate the relationship between CART and breeding of hen at the molecular, to provide theoretical basis for the regulation of poultry breeding in further. The main results of the experiment are as follows:(1)Found CART was expressed in hen ovary by using RT-PCR, and obtained230bp sequences through gene cloning. Compared with cow, swine and human, it showed that there was high homology among them, it was highly conservative in different species.(2) Immunohistochemical results showed that CART peptide was expressed in granular layer cells and theca cells of<1-2mm diameter follicle and<2-4mm follicles, and the expression in theca cells is higher than granular cells.(3) Real-time PCR showed that the expression of CART mRNA in theca cells was higher relatived to in granulosa cells in all the follicles, which may related to produce estrogen membrane in theca layer.(4) Relative abundance of CART mRNA in granulosa cells of6-8mm follicle is significantly higher than in9-12mm follicle、F5、F4、F3、F2and4-6mm follicle, the expression of CART mRNA in granulosa cells was increased in F1relatived to9-12mm follicle. F5and F2, the reason may be6-8mm follicle is the turning point either to atresa or ovulation, it imply CART gene has key effect in the key period.(5) Relative abundance of CART mRNA in theca cells of4-6mm follicle is significantly higher than in6-8mm follicle, F1、F2、F3、F4and F5and9-12mm follicle. Perhaps it was related that CART was negative for estrogen secretion regulation, and the theca layer was the important source of estrogen.