血清孕酮水平检测在克隆胚胎移植受体牛的筛选及妊娠诊断中的应用  

Serum Progesterone Level Detection for the Screening of Recipient Cattle for Cloned Embryo Transfer and Their Pregnancy Diagnosis

作　　者：贾晓[1,3] 邱瑾 舒娟 李华[1,3] 习书斌 曾溢滔[1,3] 曾凡一[1,2,3,4] JIA Xiao;QIU Jin;SHU Juan;LI Hua;XI Shu-bin;ZENG Yi-tao;ZENG Fan-yi(Shanghai Institute of Medical Genetics,Children’s Hospital of Shanghai,Children’s Hospital Affiliated to Shanghai Jiaotong University,Shanghai 20040,China;Shanghai Tao Tao Transgenic Engineering Co.,Ltd.,Shanghai 201604,China;Key Laboratory of Embryo Molecular Biology,Ministry of Health,Shanghai Key Laboratory of Embryo and Reproduction Engineering,Shanghai 20040,China;Department of Histoembryology,Genetics&Development,Shanghai Jiaotong University College of Basic Medical Sciences,Shanghai 200025,China)

机构地区：[1]上海交通大学附属儿童医院,上海市儿童医院,上海医学遗传研究所,上海200040 [2]上海滔滔转基因工程有限公司,上海201604 [3]卫生部医学分子生物学重点实验室,上海市胚胎与生殖工程重点实验室,上海200040 [4]上海交通大学基础医学院组织胚胎学与遗传发育学系,上海200025

出　　处：《中国生物工程杂志》2020年第7期1-8,共8页China Biotechnology

基　　金：国家重点科学研究计划(2014CB964701、2014CB964703);国家自然科学基金(31871484);上海市重中之重重点学科项目(2017ZZ02019);上海市自然科学基金(16ZR1428600)资助项目。

摘　　要：目的:孕酮(progesterone,P4)作为一种生殖激素,在牛的发情期和妊娠期呈规律性变化,且在妊娠的建立和维持过程中发挥着重要作用。拟应用孕酮浓度测定辅助人工观察筛选克隆胚胎移植受体牛并监测其整个妊娠过程。方法:通过对自然配种牛不同生殖阶段血液中孕酮水平的分析,建立其在发情初期、妊娠期孕酮浓度的变化规律,以此为依据辅助人工观察筛选适合胚胎移植的受体牛。同时将在体外培养7天后的体细胞核移植重构囊胚移植到所筛选出的同期发情的受体牛子宫内,并应用孕酮测定监测其妊娠状态。实验结果:(1)运用孕酮检测筛选克隆胚胎移植受体牛时,当孕酮浓度在发情第0天和第5天分别为≤0.64nmol/L和2~8nmol/L时,适宜作为胚胎移植受体牛,根据此筛选指标能够排除50%左右假发情牛。(2)运用孕酮检测较传统的人工观察方法,胚胎移植的克隆牛出生率提高了7.1倍。结论:运用牛血清孕酮检测方法能有效提高母牛生理周期判断的准确性,有利于选择合适的胚胎移植受体牛,既能实时、有效地监测怀孕受体牛的妊娠状态,又避免了靠人工观察受体牛返情、流产时的人为疏漏和误判,有效地提高受体牛的利用率,提高克隆牛的生产效率,并能推广应用于畜牧行业牛的生产繁育中。Objective:Progesterone(P4),as a reproductive hormone,changes regularly in the estrus cycle of mammals,and plays an essential role in the establishment and maintenance of pregnancy in cattle.This study through monitoring serum progesterone concentration in cattle,in addition to the routine manual estrus observation,screens for embryo transfer(ET)recipient cattle,as well as for the monitoring of reproductive status during the whole pregnancy of cattle upon Somatic Cell Nuclear Transfer(SCNT)experiments.Methods:Through the study of serum progesterone levels of natural-breeding cattle at different reproductive stages,the reference indexes of progesterone concentration at early estrus and during pregnancy were established.Based on these indexes,the recipient cattle suitable for embryo transfer were screened and selected by complementing manual estrus observation.At the same time,reconstructed blastocyst from SCNT cultured in vitro for seven days was transferred into the uterus of selected estrus-synchronized recipient cattle.The pregnancy state was monitored with serum progesterone level.Experimental results:(1)When using serum progesterone level to screen for SCNT embryo transfer recipient cattle,i.e.progesterone concentration at 0 d(≤0.64 nmol/L)and 5 d(2-8 nmol/L),more than 50%of pseudo-estrus cows can be excluded from the embryo transfer recipient list.(2)Birth rate from embryo transfer of cloned embryos through the progesterone monitoring is 7.1 fold higher than using the traditional monitoring method alone.Conclusion:The application of bovine serum progesterone monitoring can effectively improve the accuracy of cow reproductive cycle evaluation.It increases the effectiveness of suitable embryo transfer recipient cattle selection,as well as the accurate judgment of pregnancy status after SCNT embryo transfer in the recipient cattle.This practice can improve the utilization efficacy of recipient cattle selection,and the production efficiency of cloned cattle.It can be extended to other applications in the embryo

关 键 词：孕酮水平 受体牛 胚胎移植 体细胞核移植/克隆牛 

分 类 号：Q819[生物学—生物工程]