Analysis of Reproduction Function Gene of Testicle in Different Size Black Cattle Based on Transcriptome Sequencing
The aim of this experiment was to analyze the transcriptome data of testicle in different size of Shan-dong black cattle,and discover the key candidate genes affecting the reproductive function of testicles,in order to explore the molecular mechanism of maintaining the reproductive function in cattle testicle.Six bulls were divided into two groups according to different size testicle for transcriptome sequencing.The differential expression multiplier values |log2(Fold Change)| ≥1 and P-value≤0.01 were used as the conditions for selecting the differential ex-pression genes(DEGs),and GO and KEGG analysis were performed to select the candidate gene of reproductive function of testicle.The CDS region of cattle candidate gene was cloned,and qRT-PCR,Western Blot,IHC were used for further confirmation.A total of 353 DEGs were obtained from the sequencing results,including 114 up-reg-ulated genes and 239 down-regulated genes in small testicle.The results of GO functional annotation found that the items related to metabolism,development,reproduction and other processes.The results of KEGG pathway enrich-ment analysis showed that DEGs were mainly enriched in Wnt signal transduction,PI3K-Akt signaling pathway a-bout reproductive function of testicle,further screening for CCN4 related to testicle reproductive function as candi-date gene to validate.The qRT-PCR and Western Blot results showed that the relative expression of CCN4 gene and protein in big testicles was significantly lower than that in small testicle.The IHC results showed that CCN4 protein was mainly positive in the areas outside spermatids(especially sertoli cells)and the relative expression in big testi-cle was significantly lower than that in small testicle.It is speculated that the CCN4 gene may inhibit sertoli cells to affect cattle testicle function.This study provides a basic data for further study of reproductive function testicle in different size of cattle.
国家科技期刊平台
NSTL
万方数据
维普
