| Ovary was one of the most important organs in reproductive system of female mammals.It could produced eggs and secreted hormone periodically under the regulation of the nervous system and the hypothalamus-pituitary-gonadal axis hormones.Follicles and corpus luteum in ovarian alternating periodically would regulate the animal's reproductive physiology.Estrogen is a kind of reproductive hormone that have a direct relationship with animal reproduction,and it could regulate mammalian estrus and sexual behavior.Estrogen is a class of steroid hormones with similar chemical structures and 18 carbon atoms in the molecule.They exert their physiological effects by binding to their receptors.Estrogen receptors(ERs)were early detected of estrogen receptors,including estrogen receptor ?(ER?)and estrogen receptor ?(ER?).ERs localize in nucleus and play their role in gene regulation by modulated the transcription of target genes.But more and more evidence showed that ERs were not enough to complete the full effect of estrogen.In some conditions,the role of estrogen is extremely fast,which means that there is a rapid non-genomic regulation of estrogen.Recent studies found that on the membrane of many types of cells,where had a kind of membrane receptor,G protein-coupled receptor30(GPR30),mediating the rapid non-genomic effects of estrogen.Previous studies in our laboratory found that GPR30 was expressed on the cumulus-oocyte complex in goat,and adding agonists or inhibitors of GPR30 to the culture medium during oocyte maturation had different effects on the maturation process of oocytes,so we concluded that GPR30 played an important role in the process of oocyte maturation.To further prove the role of GPR30 in goat oestrus cycle and oocyte maturation,this study intended to detect the expression and localization of GPR30 during different estrous cycles in goat ovaries and investigate the expression patterns of GPR30 in follicles,corpus luteum,and atresia follicles in different development stage,whichwould be a foundation for the study on the role of GPR30 in ovarian cycle regulation,follicle development and oocyte maturation.In this study,paraffin sections and H.E.staining were used to observe the external structure of the ovary to determine the estrous cycle of the goat's ovaries.The expression and localization of GPR30 in goat ovaries,follicles,corpus luteum and atresia follicles in different stages of the estrus cycle were detected by immunohistochemistry.GPR30 m RNA expression levels in different stages of follicles development were detected by Real-Time PCR technology.The results showed that:(1)According to the development of follicular and corpus luteum in ovary and the results of H.E.staining,we demonstrated that the external structures in different stages of estrus cycle were different,and the estrous cycle stage of the ovary could be initially determined on the basis of this result.(2)The immunohistochemistry results showed that GPR30-positive products were widely distributed in goat ovaries in different stages of estrus.GPR30 was mainly located in follicles and corpus luteum.(3)GPR30 was expressed in different stages of follicle development,and was mainly located in granulosa cells,cumulus cells,oocyte cell membrane and cytoplasm.The expression of GPR30 mRNA in primordial follicles was the lowest.With the growth and development of follicles,the expression of GPR30 m RNA gradually increased,and reached the highest level in mature follicles.(4)GPR30 was positively expressed in the whole corpus luteum in the goat ovaries during diestrus.There were no significant differences in the GPR30 expression in corpus luteum in different stages of development.GPR30 was mainly located in the cytoplasm of luteal cells.The expression of GPR30 in the whole corpus luteum was diffuse.(5)GPR30 was expressed in oocytes and granulosa cells of atresia follicles.Compared with normal follicles,the positive expression of GPR30 in granulosa cells and oocyte of atresia follicles were lower than normal follicles. |
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