Analysis Of The Key Genes Expression Pattern Of Arginine-polyamine Metabolism Pathway In Pre-hierarchical Follicles Of Geese

Geese follicular development involves complex biological processes interacting with each other,and directly affects the production performance of geese.To study follicular regulation mechanism will help in improving geese laying performance and hence the development of geese industry.In recent years,studies have shown that arginine-polyamine(Arg-PA)metabolic pathways of key genes have a regulatory role on the geese follicle development.In order to understand the pathway Arg-PA specific mechanism,biological information and function,this study focuses on geese pre-hierarchy follicles,to determine the expression and analysis of the biological functions of related genes.Test sequence analysis of the geese pre-hierarchical follicles by the RNA-Sequence,was used to determine the Arg-PA pathway and the related genes relative expression,and to identify the transcriptome results from RNA to protein level by the RT-PCR and Western Blot.The Analysis of metabolic pathway for Arg-PA and the related genes functions combined with the changes of follicle development,proved the theoretical basis for the follicular development of geese.(1)Through the observation of pre-hierarchical follicular structure of geese by the HE staining,the results showed that the follicle structure with the ovarian follicle development had a change.The thickness of follicle rete and granulosa cell rete have significant difference in the pre-hierarchical follicular structure of geese(P < 0.05)However,the thickness change trend was the same,first increased and then decreased,in the middle white follicles was the largest.(2)The results also showed that the target genes;ODC,ARG,OAZ2,ODC,NOS and SMS,SRM,SMOX RNA existed in all pre-hierarchical stages of geese follicles,and had the same expression trend with the transcriptome.(3)The results of qRT-PCR showed that there were significant differences in the expression of genes related to Arg-PA pathway in the pre-hierarchical follicle of geese.ARG,ODC,NOS and SMS had the highest expression in primary follicles than ADC and SMOX respectively.ADC have lowest expression in small white follicles;the SMOX have the highest expression in the middle white follicles but the genes of ARG,ODC,NOS,SRM and SMS have the lowest,the genes of OAZ2 had lower expression in large white follicles.The genes had significant difference between the highest and lowest expression of mRNA(P<0.05).(4)Protein expression analysis of ODC,ARG and NOS by Western Blot results showed that ARG and NOS protein had the highest expression in primary follicles,and the lowest expression in middle white follicles,ODC protein had the highest expression in large white follicles and the lowest expression in small yellow follicles,there was a significant difference between the highest expression and lowest expression(P<0.05).(5)The expression analysis of genes related to Arg-PA pathway with follicular development suggested that the Arg-PA pathway may be regulated at initial stage of primordial follicle growth,the genes of ARG,ODC,NOS,SMS and SRM can promote the follicular development by regulating the granulosa cells proliferation and development,the genes of ADC,OAZ,SMOX inhibit follicular development.