利用微陣列技術(shù)研究豬卵泡發(fā)育相關(guān)基因的差異表達.pdf

1、中國農(nóng)業(yè)大學(xué)博士學(xué)位論文利用微陣列技術(shù)研究豬卵泡發(fā)育相關(guān)基因的差異表達姓名：師科榮申請學(xué)位級別：博士專業(yè)：動物遺傳育種與繁殖指導(dǎo)教師：吳常信李寧20060601ABSTRACTInordcrtoresearchmoleculardevelopmentalmechanismsofmammalianovarianfollicle，porcineovarianfollicleswereselectedasrepresentativeresea

2、rchtargetsTheobjectiveofthestudywastofindovarianfolliclesdevelopmentrelatedgenesbyclarifyingtheexpressionprofilesofdifferentdevelopmentalstagefolliclesTheattentionsofthestudywerefixedonthreekindsofrepresentativeovarianfo

3、lliclesinthewholefollicledevelopmentprocessTheywereprimordialfollicles(namedasP)，middledevelopmentalstagefollicles(specialtoPre—antralfollicles，namedasM)andlastdevelopmentalstagefollicles(specialtoLargeantralfollicles，na

4、medasL)Afterbeingisolatedfromovariesofprepubertalgilt，RNAprobesderivedfromextractedRNAwerehybridizedwithcommercialoligomicroarrays，andafteraseriesofdataanalysis，weobtainedthedifferentiallyexpressedgenesthatrelatedtothede

5、velopmentofovarianfolliclesResultsonbothfolliclesisolationandgenesrelatedtoovarianfollicledevelopmentwereasfollows：(1)Basedonthepreviousmethods，thestudyimprovedtheyieldofprimordialfolliclesbymeansofincreasingthelayernumb

6、ersaftercentrifugationinPercollgradientsTheisolatedprimordialfolliclesfromaprepubertalovarywereimprovedfrom185x10’21x105to779x105Andimmunoinfluenrenceanalysisshowedthataveragely9831％O73％ofcellsthatwithrelativelylargersiz

7、einfollicularseparationswasprimordialfolliclesSo，notonlyhigh—yieldedbuthighlypurifedprimordialfolliclescouldbeisolatedbyusingthemethodadvancedinthestudy(2)TheGiemsaandHoechst33342stainingmethodcouldbeusedtodistinguishpri

8、mordialfolliclesfromcontaminatedsomaticcellsAndespeciallytheMaY2couldbedevelopedasarecognizingmarkerofprimordialfoLLiclesbecauseofitsprecisionandeKactness(3)Becauseofsomanyconnectivetissuesinporcineovarymechanicalmethods

9、suchasmicro—dissectionorcuttingmethod，weremoresuitableforisolatingpre—antralfollicles(1004009m)thanothercomb—scrapeandenzymaticmethodThelatertwomethodshadlowerfollicleyieldf2～4folliclesperovary)andquality(4)Thegenomescal

10、edexpressionprofileofP(primordialfollicles)wasgreatlydiffi!rentfromthatoftheothertwokindsoffollicles(MandL)Andthelatertwodevelopmentalstagefollicleswerehighlycorrelatedeachotherwiththebiologicalcorrelationcoefficient(R2)

11、ofO892Allthefeaturesofexpressionpattemsshowedthatagreatmechanismregulatedtheprimordialfolliclepoolkeepquiescentintheovaryandadeepandwideinternalchangeresultedininitiationtheprimordialfolliclesintogrowth(5)Somedistributio

12、ncharacteristicsofdifferentiallyexpressedgenessuggestedthatovarianfolliclesinitiatedinaphysiologicalcyclecouldgrowintolargeantralfolliclesinageneralsalTleregulationpatIem，onlywithsomesubtlemetabolismpathwaysindifferentde