| Kidding rate of female goats is determined by ovulation rate,which is mainly affected by genetic factors.As the main functional carrier of kidding rate,the ovary mainly contains ovarian stroma and follicles at different stages of development.Ovarian stroma can not only affect the formation of follicles,but also transfer hormones and nutrients to follicles,so as to jointly affect the reproductive performance of female animals.At present,the research on the whole ovary was very common,but the differences between ovarian stroma and follicles have not been reported yet.Chuanzhong black goat is an ideal model to study the reproductive characteristics of goats because of its outstanding reproductive performance and strong adaptability.In order to study the changes of transcriptome level in different parts of ovary,explore the genetic mechanism of ovarian stroma and follicle,and screen the potential candidate molecules that can be used to improve the ovulation rate of goats,this study took the ovary of Chuanzhong black goat in estrous period as the research object,and used RNA-seq and Small RNA-seq to analyze the expression profile of messenger RNA(mRNA)and microRNA(miRNA)in ovarian stroma,large follicle and small follicle,then the differential expression,functional analysis and conjoint analysis were carried out.The main results are as follows:(1)In this study,10 healthy Chuanzhong black female goats aged 3.5-4 were selected,slaughtered after estrous treatment at the same time,collected ovarian stroma,and separated follicles under microscope.Each goat got 1-2 large follicles(d > 10 mm)and 8-15 small follicles(d < 3 mm)on average.(2)RNA-seq data analysis showed that 2918 and 622 genes were differentially expressed in OS vs LF and OS vs SF respectively,277 of them were differentially expressed in this double comparison.Functional analysis showed that DEGs of OS vs LF were mainly enriched in the biological processes of steroid metabolism,enzyme transport activity and signal pathways of steroid biosynthesis and cAMP signaling;DEGs of OS vs SF were mainly enriched in the biological processes of cell cycle process,receptor activity and signal pathways of cell cycle,oocyte meiosis;cell adhesion molecules,ovarian steroidogenesis were enriched differently in the double comparison(OS vs LF,OS vs SF).(3)Small RNA-seq results showed that 32 and 16 miRNAs were differentially expressed in OS vs LF and OS vs SF,respectively,and 11 miRNAs were differentially expressed in the double comparison.The functional analysis of target genes of DEmiRNAs showed that OS vs LF was mainly enriched in the biological processes of cell surface receptors,tissue development and signal pathways of MAPK signaling pathway,cholesterol metabolism;OS vs SF was mainly enriched in biological processes such as cell differentiation regulation,transcription factor activity,Notch signaling pathway and ovarian steroidogenesis signaling pathway;TGF-β signaling pathway and cell adhesion molecule signaling pathway were enriched in the double comparison(OS vs LF,OS vs SF).(4)Combined analysis of Small RNA-seq and RNA-seq showed that PTCH1,CLDN10,miR-122 and miR-10a-5p were highly expressed in ovarian stroma,may play an important role in ovarian organogenesis and hormone secretion;GRIA1,CYP11A1,miR-202-5p and miR-543-5p were highly expressed in large follicles,may play an important role in follicular maturation;BUB1B,CCNE1,miR-33b-3p and miR-1185-3p were highly expressed in small follicles,may play an important role in follicular development.(5)The results of transcriptome sequencing showed that the expression patterns of gene and miRNA in ovarian stroma,large follicle and small follicle were tissue space specific. |
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