Profiling Of IncRNA And MRNA In Goat Ovary At Different Estrus Stage

Goat is an important multi-purpose livestock and is an important farm animal,its fecundity is one of the important links of goat breeding industry,and the reproduction is mainly regulated by reproductive hormones.The ovary is the main reproductive organ and directly mediates ovulation and the secretion of estrogen,which plays a key role in the reproductive performance of goat.In recent years,it has been found that more and more lncRNAs are involved in the regulation of mammalian breeding.As a protein-encoding RNA,the role of mRNA in mammalian ovarian function has also received increasing attention from researchers.However,few studies have studied the effects of lncRNA and mRNA in goat ovary on goat breeding.In this study,we collected a total of 10 goat ovaries（5 luteal ovary and 5 follicular ovary）.RNA-seq technique was used to perform genome analysis of the luteal phase and follicular goat ovary.Through the methods of co-localization and co-expression analysis,the related genes of lncRNA are predicted.Functional analysis of GO and KEGG for its related gene and mRNA gene were performed to screen the long non-coding RNA and mRNA that related to goat breeding ability,and to study the transcriptional differences of goat ovary in different estrus cycles.This study can provide theoretical basis for studying goat breeding.（1）RNA-seq was performed on the ovary of the luteal phase and the follicular phase from Anhui white goat.In this sequencing,we have established 10 libraries.the luteal ovarian（LO）and follicular ovary（FO）libraries obtained 106510454,108716574,97775106,104110076,96440340,146672958,145511932,116846442,139588328,101972338 raw reads,respectively.A total of 1164144548 raw reads were obtained.After filtering out the adaptor sequences,empty sequence,and low-quality sequences,the luteal ovarian（LO）and follicular ovary（FO）libraries obtained 104409940、103650760、93334318、99345656、91918954、141469312、140170314、112863548、135618434、99232876 clean reads,respectively.A total of 1122014112 clean reads were obtained,（2）The coding potential analysis software such as CNCI,CPC,Pfam-sca were used to identify the lncRNA and mRNA,4926 lncRNA,1454 TUCP and 43766 mRNAs were identified for further analysis.115/22/3770 differentially expressed lncRNA/TUCP/mRNA were screened in the luteal and follicular ovaries.We predicted lncRNA/TUCP-related mRNA genes by co-localization and co-expression analysis methods.Based on the correlation between the differential lncRNA/TUCP and gene,we predicted 2584/904 related genes;based on the positional relationship between the differential lncRNA/TUCP and the gene,we predicted 326/73 related target genes.（3）In this study,the above predicted genes were used for GO and KEGG enrichment analysis.The results showed that lncRNA/TUCP,which is highly expressed in the ovary of luteal phase goats,is mainly related to progesterone synthesis.lncRNA/TUCP,such as XR₀01918177.1,TUCP₀01362 may regulate progesterone synthesis;lncRNA/TUCP,which is highly expressed in ovarian follicles,is mainly related to oocyte growth and maturation,and we screen out the lncRNA/TUCP that may regulated the growth and maturation of oocytes,such as XR₀01917388.1,TUCP₀00849 and so on（4）A total of 3770 differential mRNAs were identified for further analysis in this study.Among them,1727 mRNAs were up-regulated in the luteal phase of the ovary,and 2043 mRNAs were up-regulated in the follicles phase of the ovary.These differential mRNAs were used for GO and KEGG enrichment analysis.Through functional analysis,we obtained some mRNAs that are highly expressed in the ovarian ovary,such as HSD17B7,3BHSD,SRD5A2,and so on may be related to the synthesis of progesterone;Highly expressed of mRNA in the ovary during follicular phase,such as RPL12,RPS13,RPL10 and so on,is associated with the growth and maturation of oocyte.（5）The 3lncRNA and 3mRNA from 10 libraries were selected randomly,and the GAPDH was used as an internal reference,the qRT-PCR was used to verify the sequencing results.The results show that the sequencing data of RNA-seq is reliable.