山羊卵母细胞生发泡染色质构型的研究-基础兽医学专业论文.docxVIP

东北农业大学农学硕士学位论文摘 东北农业大学农学硕士学位论文 摘 要 (近年来哺乳动物卵母细胞体外成熟技术已取得长足的进展，但还存在很多问题没有 解决。尽管在成熟培养前对卵丘一卵母细胞复合体(COCs)进行严格的选择，但是即使从 同一个卵巢收集的卵母细胞其第一次减数分裂前期双线期的时相也各不相同，因而从不 同个体的卵巢、不同大小的卵泡内收集的卵母细胞实际上是异质的。这很可能是卵母细 胞IVM质量差异较大，造成体外胚胎发育能力差的原因之一。)本实验应用荧光显微镜研 究了山羊不同大小、健康程度卵泡来的和体内、外发育不同时间卵母细胞生发泡构型以 及在此期间PdqA合成情况的变化规律。实验结果表明： 1采用活体荧光染料Hoechst 33342对山羊生发泡进行观鸯‰根据染色质构型及核 仁大小将其分为8种形态：GV0、GVl、GV2(GV2．1、GV2．2)、GV3(GV3．1、GV3．2) GV4和GVBD．／)一声。～一一 2腔前卵泡卵母细胞生发泡都为GV0期；直径为0．5．0．8mm卵泡卵母细胞都为GVI 期：随着卵泡的发育，GV3期卵母细胞比例明显增高。 3体外成熟过程中，随着培养时间的延长。GVI、GV2及GV4的比率有所下降， 而GV3和GVBD的比率明显上升。 4健康卵泡中GV3的比率显著高于闭锁卵泡，而闭锁卵泡中GV4和GVBD的比率 显著高于健康卵泡。 5随着卵泡直径增大，卵母细胞核仁直径逐渐变小。 6体内注射FSH和hCG后的排卵前卵泡卵母细胞生发泡染色质构型主要为GV3， 其中GV3-l占绝对优势，而GV3．2的比率较低。 7直径为0．5-0．8mm的卵泡卵生发泡内RNA大量合成，1-3mm卵泡中RNA合成较 减弱，而3ram卵泡内卵母细胞RNA合成结束。 关键词：山羊；卵母细胞；生发泡：染色质构型：核仁、 Studies Studies on Germinal Vesicle Chromatin Configuration in Goat Oocyte Abstract AlthOUgh techniques of in vitro maturation(IVM)of mammalian OOCytes have made great progress，some problems are still unsolved．Even thorIgh COCs were strictly selected before in vitro culture，the c}Lromatin configuration of dietyate stage OOCytes Was different． Therefore．oocytes collected from follieles of different diameters in the same ovary were heterogeneous This is one of the reasons for tIle poor quality of in vitro matured OOCytes．In this study,we observed the changes of germinal vesicle cIlromatin configuration in goat OOCytes from follicles of different diameter and health status，and during in vitro and vivo maturation using fluorescence microscopy． The results are as follows： 1．Eight tvpes of GV chromatin configurations were observed with Hoechst 33342 staining in the goat oocytes according to the morphology of cllromatin and the size of nucleolus and they were GV0、GVl、GV2(GV2—1、GV2-2)、GV3(GV3-1、GV3—2) GV4 and GVBD。 2．Ooc”es from the pre—antral follicles were all at GV0 stage and OOCytes from 0．5-0．8mm follicles were all at GVl stage．With the follicular development,the proportion of GV3 stage was significantly i