Effect Of Mixed PMSG/FSH Treatment On Ovarian Response And Transcriptome Sequencing Of Ovarian Tissue In Estrus Synchronized Dairy Goats

In order to improve the reproductive rate of dairy goats,explore the efficient and practical reproductive technology of mixed use of follicle-stimulating hormone(FSH)and pregnant horse serum gonadotropin(PMSG)under the condition of estrus during the same period.In this study,100 Xinong Saanen ewes were randomly divided into four groups,each with 25 ewes.They are mixed treatment 1 group of PFM1: PMSG 200 IU + FSH 60IU(1:0.3);mixed treatment 2 groups of PFM2: PMSG 200 IU + FSH 120 IU(1:0.6);single use of PMSG group PC: PMSG 200IU;single use of FSH Group FC: FSH 120 IU.The main results are as follows by measuring the same period of estrus,hormone levels,ovarian morphology,ovarian response and ovarian transcriptome sequencing:1.The effect of PMSG/FSH mixed treatment on estrus and hormone levels of dairy goats at the same timeThe estrus effect of the four groups of hormone treatment programs during the same period is better,reaching more than 90%.The best estrus rate at 72 hours was in the FC group,reaching 96.15%,and the worst was in the PC group,reaching 92%.The mixed treatment was in the middle,and the PFM2 group(95.83%)was better than the PFM1group(92.59%).However,the FC group showed the latest estrus start time(44h),and the PC group had the shortest estrus duration(19h).PFM2 showed the earliest estrus after thrombus removal,and the estrus lasted the longest,which is beneficial to breeding and has the best estrus effect.The plasma reproductive hormones FSH,LH,E2,and P4 of the four groups of hormone treatment programs have no significant difference in the estrus stage.In the two groups of mixed treatment,the plasma reproductive hormones have a more obvious peak trend change,indicating that the pituitary gonad axis is more sensitive to the mixed hormone stimulation and the reproductive physiological activities are more intense.2.The effect of PMSG/FSH mixed treatment on ovarian morphology and ovarian response in dairy goatsThere were no significant differences in the length and width of the ovaries and the morphological development of various types of follicles in the dairy goats of the four groups of hormone treatment programs,indicating that the doses and proportions of the four groups of hormones did not cause damage to the basic physiology of the ovary.The four groups were treated with hormones to observe the ovarian response and recover embryos.Compared with the FC group,the ovarian response of the PFM2 group was stable and the use efficiency was higher.Compared with the PFM1 group and the PC group,the PFM2 group had more ovulation points and no large follicles.The ovarian response group performed best,which reduced the adverse effects of using PMSG and was beneficial to the fertilization process and early embryo development.3.Transcriptome sequencing and analysis of milk goat ovarian tissue treated with PMSG/FSH mixedThe mixed treatment of PMSG/FSH and the treatment of PMSG and FSH alone resulted in 109 and 751 differential genes,respectively.After enrichment analysis of different genes by GO and KEGG,significant GO items related to ovarian development and reproduction include angiogenesis,signal transduction,transmembrane signal transduction,receptor activity,and binding;KEGG pathway has cell adhesion molecules(CAMs),Ovarian steroid production,endocytosis,PI3K-Akt signaling pathway,glycosphingolipid biosynthesis,tight junctions,adhesion junctions,etc.Five genes(PROM1,EGR4,EPCAM,NR4A2,BTG2)related to ovarian cell development and hormone transmission were selected as key candidate genes.In summary: PMSG/FSH mixed treatment has a positive effect on the estrus effect and hormone level at the same time.PFM2 shows estrus earliest after thrombus removal,and the estrus lasts the longest,which is beneficial to breeding and has the best estrus effect.Compared with the PFM2 group,the ovarian response has more ovulation points and no large follicles.The performance of the four groups is the best,which reduces the adverse effects of using PMSG and is beneficial to the fertilization process and early embryo development.In general,the mixing ratio of PFM2 is suitable for the treatment of estrus hormones in the production of dairy goats at the same time.Transcriptome sequencing screened to obtain 5 key candidate genes related to ovarian cell development and hormone conduction(PROM1,EGR4,EPCAM,NR4A2,BTG2).Compared with PC group and FC group in PFM2 treatment group,it promoted PROM1,EGR4,EPCAM,etc.The expression of genes related to the development of ovarian luteal cells and the secretion of progesterone,inhibits NR4A2,BTG2,etc.and prevents the expression of genes related to the differentiation of follicular granulosa cells into luteal cells,so that the ovaries have more ovulation points and no large follicles,which is beneficial to fertilization and early embryos development.These genes can be followed up for functional verification,laying the foundation for the regulation mechanism of ovarian tissue-related gene reproduction after PMSG/FSH mixed treatment.