The DGE Selection And Gene Function Analysis Controlled By Nutrition Level Of Estrous Stall-feed Sheep In Non-breeding Season

The seasonal breeding is one of the main bottlenecks in limiting the sheep reproduction effectiveness which are typically seasonal estrous animals.At present,it is generally accepted that the seasonal oestrus sheep this characteristic mainly is regulated by the light path,in view of the characteristic people preliminarily determines the access control model of light.But in actual production,in the breeding season of feeding high nutrition level of kazak sheep have appeared in the rutting phenomenon,this phenomenon shows not only has the light cycle to sheep estrous play a key role,and nutrition of sheep rutting performance also has certain effect,so the nutrition level induced estrus mechanism and regulatory mechanism of photoperiod whether consistent? For the study of sheep production has great guiding significance.Therefore,this research through digital gene expression spectrum filter induced by nutrient levels change kazak sheep breeding season heat and lack of feeling of differentially expressed genes in the hypothalamus,pituitary,ovary,related bioinformatics analysis,and the screening of differentially expressed genes related to estrus for functional verification.Established under the condition of different nutrition levels of kazak sheep,sheep in the breeding season to observe and record test heat condition.Slaughtered respectively,lack of affection season three estrus(EN),three estrus(just)(ENN)and the lack of affection(AN),a ewe,obtained the hypothalamus pituitary(P)(H),and ovaries(O),a total of nine DGE building library,through bioinformatics analysis.Screening differentially expressed genes between them,and then GO analysis and statistical analysis of KEGG pathways.Results show that the lack of seasonal oestrus,estrus(just),and the sheep of the situation of lack of hypothalamus tissues than to the reference gene sequence of article 2925936,2956401,2647042,respectively;Lack of seasonal oestrus,estrus(just)and the sheep of the situation of lack of pituitary and reference gene sequence alignment with article 3198493,3327125,3635448;Lack of seasonal oestrus,estrus(just)and the lack of feeling of sheep ovarian and reference gene sequence alignment with article3757637,3694199,3135565;Lack of seasonal oestrus,estrus(just)and lack of affection of the hypothalamus of differentially expressed genes,respectively,in 808,1906,spent seasonal oestrus,estrus(just)and spent 552 differentially expressed genes in the affection of the pituitary gland,1091,spent seasonal oestrus,estrus(just)and lack of love in the ovaries were 58,201 differentially expressed genes,differentially expressed genes related to heat there were 13,11,5,4,3,and 4,containing the Kiss1,CNH2 and GNAQ,MEK1 genes.Randomly select several differentially expressed a higher amount of real-time fluorescent quantitative PCR gene,its expression trend basic consistent with the results of sequencing,illustrate the sequencing results are true and correct.And then GO on them and KEGG pathway analysis showed that GNAQ differentially expressed genes in multiple pathways play a role,especially in GnRH signaling pathways,GNAQ,the downstream pathway GnRH receptor,is likely to be indirect Gn RH to influence the action of heat.Through Kiss1 and GNAQ quantitative study of gene expression in sheep,kiss-1 as important genes in various organizations in the light path are expressed,and express the amount ishigher,and GNAQ only in the spleen,liver,and the high amount of expression in the hypothalamus,other organization express the amount is very low.By constructing pcDNA3.1(+)-GNAQ expression vector,transfection in ovarian granulosa cells,the results did not find under laser excitation fluorescence,showed GNAQ were not expressed in ovary,remains to be studied.Through the analysis of genes and the related heat pathways,may the breeding season sheep estrous and tyrosine,folic acid and glycine,and other nutrients,tyrosine pathways of MEK1 gene by GnRH pathways in FSH and LH;Folic acid may swallow a role within the pathways through prompt estradiol and impact of estrus;Glycine by glycine,serine and threonine metabolism play a role in;They work together to influence the estrous.