利用微阵列技术研究猪卵泡发育相关基因的差异表达-动物遗传育种与繁殖专业毕业论文.docxVIP

ABSTRACTIn ABSTRACT In ordcr to research molecular developmental mechanisms of mammalian ovarian follicle，porcine ovarian follicles were selected representative research targets The objective ofthe study was to find ovarian follicles development related genes by clarifying the expression profiles of different developmental stage follicles The attentions of the study were fixed three kinds of representative ovarian follicles in the whole follicle development process They were primordial follicles(named P)， middle developmental stage follicles(special to Pre—antral follicles，named as M)and last developmental stage follicles(special to Large antral follicles，named L)After being isolated from ovaries of prepubertal gilt，RNA probes derived from extracted RNA were hybridized with commercial oligo microarrays，and after series ofdata analysis，we obtained the differentially expressed genes that related to the development of ovarian follicles．Results both follicles isolation and genes related to ovarian follicle development were follows： (1)Based the previous methods，the study improved the yield of primordial follicles by means of increasing the layer numbers after centrifugation in Percoll gradients．The isolated primordial follicles from prepubertal ovary were improved from 1．85x10’-2 1x105 to 7．79x105．And immunoinfluenrence analysis showed that averagely 98 3 1％±O 73％of cells that with relatively larger size in follicular separations was primordial follicles．So，not only high—yielded but highly purifed primordial follicles could be isolated by using the method advanced in the study． (2)The Giemsa and Hoechst33342 staining method could be used to distinguish primordial follicles from contaminated somatic cells．And especially the MaY2 could be developed recognizing marker ofprimordialfoLLicles becauseofits precisionand eKactness． (3)Because of many connective tissues in porcine ovary,mechanical methods．such micro—dissection cutting method，were more suitable for isolating pre—a