摘要
取发育早期日本沼虾(Macrobrachium nipponense)的精巢,剪碎,分散成细胞悬液,用离体细胞培养法进行体外培养,培养基中不含17α-甲基睾酮(17α-MT)的为对照组,设定3种不同质量浓度水平组,分别为300ng/mL、1500ng/mL和3000ng/mL,测定生精细胞的体外存活率并观察生精细胞体外培养的生长行为和分化情况。结果表明,3种浓度梯度下生精细胞的存活率没有显著差异,17α-MT的质量浓度为1500ng/mL组分化为精子细胞四分体的数量和长出棘突的精细胞数量均比对照组和其他实验组多,差异极显著(P<0.01)。初步确定生精细胞体外培养过程中添加不同质量浓度的17α-MT不影响细胞的存活率,但影响生精细胞的体外分化,17α-MT的适宜质量浓度为1500ng/mL。
17α-MT has important roles in the regulation of many physiological,developmental,and behavioral processes in aquaculture and medicament.Up to now,the effects of 17α-MT on the differentiation of spermatogenic cells from Macrobrachium nipponense in vitro have seldom been studied.In this study,M.nipponense were used as research material,cells in testis were isolated and cultured in vitro.In order to evaluate the survival rate,growth state, and differentiation of the spermatogenic cells,the culture medium was supplemented with 17α-MT at 0 ng/mL,300 ng/mL,1 500 ng/mL,and 3 000 ng/mL respectively.The results showed that during the process of meiosis minority of spermatocytes formed a tetrad of spermatids in vitro.During cell division,the plasma membrane did not separate thoroughly,but formed a synplasm.Spermatids in the early phrase could finish partial differentiation and some filament appeared,whereafter,a round bright acrosome vesicle inside the cell emerged.The vesicle became larger,squeezing cytoplasm and nuclear to the other side.Spike arose from minority of spermatids.There was no significant difference on survival rate of spermatogenic cells at different concentrations of 17α-MT（P〉0.05）,but the number of spermatids differentiated was obtained at 17α-MT concentrations of 1 500 ng/mL rather than other three groups（P〈0.01）.It is concluded that 17α-MT has no effect on survival rate of spermatogenic cells in vitro;the concentration of 1 500 ng/mL 17α-MT supplemented to culture medium is the suitable hormone concentration for culturing spermatogenic cells in vitro.So it was suggested that once entering inside the cell the steroid hormone would bind with a specific receptor which was found only in the cytoplasm of the target cell.The receptor bound steroid hormone then travelled into the nucleus and bound to another specific receptor on the chromatin.
出处
《中国水产科学》
CAS
CSCD
北大核心
2009年第2期289-294,共6页
Journal of Fishery Sciences of China
基金
国家自然科学基金资助项目(30371115)
河北大学青年基金资助项目(2005Q13)
