摘要
以小鼠２－细胞胚为材料，研究电融合请参数。以Ｍ２液为融合液时，除在０．３１ｋＶ／ｃｍ，１２８０ｓ，１次脉冲条件下获１００％融合率外，其他条件均造成较多的胚胎溶解。而持续时间在１０～１２８０ｓ范围内，以０．３ｍｏｌ／Ｌ甘露醇液为融合液，电场强度为０．３１～２．０４ｋＶ／ｃｍ条件下；及以０．２６ｍｏｌ／Ｌ蔗糖液为融合液，电场强度为０．３１～１．４１ｋＶ／ｃｍ范围内时可获较高的融合率。实验表明：在０．４７ｋＶ／ｃｍ电场条件下，蔗糖液与甘露醇液的渗透压对融合率无显著的影响；电场强度为２．０４ｋＶ／ｃｍ，以高渗甘露醇液（０．３０ｍｏｌ／Ｌ）为融合液时，胚的融合率比等渗甘露醇液（０．２５ｍｏｌ／Ｌ）和低渗甘露醇液（０．２０ｍｏｌ／Ｌ）做融合液时的融合结果高，而胚的死亡率低。融合后的胚与对照２－细胞胚都可继续发育．４８ｈ后发生致密化，此时融合胚发育到４－细胞，对照组为８－细胞。根据０．２６ｍｏｌ／Ｌ蔗糖液做融合液时的结果，绘制一条获高融合率时电场强度与脉冲持续时间的关系曲线。
The vertebrate ocular lens has been proved to a good experimental model for r studying many problems of development,such as the effects of drugs on development,formation and distribution of gap junctions,and metabolic cooperation.In this paper,we used monoclonal antibodies,termed NC6 and ND6,specific for gap junction protein,as a probe to study the time course of effect of antibodies on development in the chicken embryonic lenses.NC6 or ND6 was injected into the right eye of chicken embryo at the 20th stage.The uninjected left eye of the same embryo was used as the control.The embryonic heads were fixed 8,12,24,36,or 48hours after injection.Lenses were removed from the eyes and measured after fixation.The size difference of lenses between injected and control groups in each time group was compared statistically.A time course curve was made with the P value from each time group.The results showed;(1)In NC6 injection,a significant size difference between injected and control lenses had already appeared in the 8 hour group(P<0.05 to 0.01)and disappeared in the 48 hour group(P>0.05).The time course curve in NC6 was smooth:(2)In ND6 injection,there was a significant difference in lens size only in the 24 hour group(P<0.05 to 0.01).The time course curve in ND6 was precipitous;(3)The peak of the time course curve was at the 24 hour point in both NC 6 and ND6 injections.The change of sensitivity to antibodies in embryos at different stages was also quantitatively analyzed this study.NC6 or ND6 was injected into eyes of chicken embryos at 20th or 24th stage.After 24 hours,the size differences of lenses between injected and control groups were statistically compared.The results showed that the lenses from embryos at the 20 th stage were highly sensitive to the antibodies(P<0.01 to 0.001 for NC6;P<0.05 to 0.01 for ND6),but the sensitivity of lenses at the 24th stage was much lower(P>0.05 or P<0.0l for NC6;P>0.05 for ND6).The difference in the sensitivity to antibodies between 20th and 24th stages is considered to be relate
出处
《解剖学报》
CAS
CSCD
北大核心
1994年第2期214-218,共5页
Acta Anatomica Sinica
基金
国家教委霍英东青年教师基金
关键词
电融合
渗透压
细胞胚
细胞融合
Development
Time course
Sensitivity
Monoclonal antibody NC6 and ND6
Chicken embryonic lens
