摘要
本试验选用纯系 C_(57)BL/6小鼠,将雄鼠脾细胞悬液经腹腔注射免疫同基因型雌鼠。加强免疫后,常规尾静脉采血制备抗 H-Y 抗血清。采用精子微量细胞毒试验和酶联免疫法(ELISA)2种方法进行抗血清检测。将特异性和效价均较好的抗H-Y 抗血清用于间接免疫荧光法鉴定小鼠胚胎性别。共鉴定桑椹胚和早期囊胚462枚,呈荧光阳性(雄性)的占47%(219/462);呈荧光阴性(雌性)的占53%(243/462),经 X^2检验,符合正常性比例(1:1,P>0.05)。将这2类胚胎分别移植给假孕受体进行验证,并对验证结果进行分析。
H-Y antisera were produced in C_(57)BL/6 female mice by repeated intraperitoneal injec- tions of syngeneic male spleen cells.Antiserum samples were taken from superimmunized fe- males by tail bleeding.Two methods were used to detect antisera.One was sperm micro cytotoxicity test,and the other enzyme-linked immunosorbent assay(ELISA).The chosen antiscra were used in an indirect immunofluorescence assay to detect H-Y antigen on murine embryos for sexing.The total number of sexed cmbryos was 462,47% of which were fluores- cent(H-Y positive)embryos,and 53% nonfluorescent(H-Y negative)ones.There was no significant difference from the normal sex ratio(1:1,P>0.05).The two classes of embryos were transplanted to pseudopregnant recipients and allowed to term.The results were ana- lysed.It showed that H-Y antigen was highly conservative and suggested the possibility to identify embryo sex of other species with mouse antiserum.
关键词
小鼠胚胎
性别鉴定
H-抗原法
H-Y antigen
anti-H-Y antiscrum
murine embryo
sexing
