| Bovine estrous cycle is divided by ovarian state into two periods, the follicular phase and the luteal phase. Reproductive Endocrinology regulates the process of the estrous cycle, leading to a series of changes of follicle formation, maturation, ovulation and corpus luteum formation. Meanwhile, steroid hormones produced by the ovaries affect the gonads, makes reproductive system to produce a corresponding change and to complete the appropriate physiological functions, such as, egg maturation, fertilization, implantation and pregnancy. Estrogen and progesterone play an important role in the estrous cycle or pregnancy process. Therefore, this study experiment on cow reproductive organs with different physiological states in order to investigate estrogen and progesterone receptors distribution change in different physiological states, providing theoretical and experimental evidence for enriching reproduction theory and improving reproductive rate. Meanwhile, oviduct cells have been already cultured in vitro, in vitro materials have been provided for further cow reproductive physiology research.Test is divided into two parts:firstly selected four sets of cow reproductive organs in a clear follicular or luteal phase then frozen. Use the RT-PCR method to evaluate distribution of estrogen and progesterone receptor change in experiment of3position (the ovary, fallopian tube and uterus) crosscut samples, providing a basis for further study of hormonal changes in the course of the estrous cycle. Secondly, oviduct epithelial cells were isolated, cultured and identified in vitro. Cells were cryopreservated and established cell bank of oviduct cells, providing materials in vitro for the future research of oviduct productive physiology.Main results are as follows:1Receptors of ER α, ERβ and PR distribute differently according to different parts of the reproductive system.In the follicular phase, ERa, ERβ and PR levels all in ovarian highest, followed by the fallopian tubes, in uterus lowest, different significantly (P<0.05). During the luteal phase, ERa is highest in the uterus, followed by the fallopian tubes, ovaries are lowest, different significantly (P <0.05);in oviduct ER(3level is highest, followed by ovarian, lowest in uterine, different significantly (P<0.05); PR is highest in the ovary, secondly fallopian tube, uterus lowest, different significantly (P<0.05).2ERa, ERβ and PR are in a dynamic change in different physiological statesOvaries and fallopian tubes in follicular phase, ERa was significantly higher than the luteal phase (P<0.05), while in the uterus, two periods of ERa unchanged (P>0.05), Ovaries in follicular phase ERβ was significantly higher than the luteal phase(P<0.05),while ERβin the uterus of luteal phase was significantly higher than follicular phase; and has no difference in the two states of oviduct (P>0.05).PR in ovaries and fallopian tubes of follicular phase was significantly higher than the luteal phase (P<0.05); in uterus, the content of PR in the follicular phase and the luteal phase did not change (P>0.05).3All parts and the physiological state, ERα expression level was significantly higher than the expression of ER β, that means:ER a is the main type of estrogen receptor.4Established cell bank of oviduct cells, cell purity more than90%,the motility was above78%after thaw.First isolated epithelial cells takes a long time to adhere, suspension time is5-7days, like to stay in a group. After adherent cells first form to colony, one week can cover the bottom. From the second generation the attaching time changes to1-1.5days, epithelial cells could be passaged to the7th generation. After the second generation, with the increase of the number of passages, the cell passaging interval extended. |
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