水蕨精子发生过程中中心体蛋白和微管蛋白的免疫荧光定位

Immunofluorescence Localization of Centrin and Tubulin during Spermatogenesis of Ceratopteris thalictroides

采用透射电镜技术和免疫荧光标记技术对水蕨精子发生的超微结构以及中心体蛋白和微管蛋白在精子发生过程中的动态表达进行了观察。研究发现:(1)生毛体分化早期周围有放射状微管分布,这与线粒体向生毛体的聚集有关。(2)免疫荧光观察表明,中心体蛋白仅定位于生毛体、基体和鞭毛带上,自生毛体至基体阶段呈现明亮的荧光标记,在核塑形、鞭毛形成至精子成熟阶段,中心体蛋白荧光标记随着鞭毛的发生而逐渐减弱,至游动精子阶段中心体蛋白荧光标记信号几乎消失。(3)微管蛋白早期荧光标记与中心体蛋白标记形相同,在生毛体、鞭毛带、基体等运动细胞器上呈现明亮荧光标记,但微管蛋白随着鞭毛的发生其荧光标记越来越强。从二者的时空表达特征可以推断,中心体蛋白主要是运动细胞器的组织者,而非这些运动细胞器的结构成分,其功能是参与或负责中心粒、基体和鞭毛的发生。

The ultrastructure and the dynamic expression of the centrin and tubulin during spermatogenesis of the fern Ceratopteris thalictroides were observed using the TEM technology and immunofluorescence technology.The present investigation shows that：（1）there are abundant radial microtubules around the early blepharoplast.The function of the radial microtubules may be used to converge mitochondria to the blepharoplast.（2）Immunofluorescence observations show that centrin is only located in the blepharoplast,basal bodies and flagellar band.The fluorescence labeling of centrin becomes brighter from the blepharoplast stage to the basal body stage.Then the fluorescence labeling of centrin becomes weak gradually with the formation of flagella and almost disappears at the spermatozoid stage.（3）The early fluorescence labeling of tubulin,showing the same form as the centrin labelling,is located in the blepharoplast,basal bodies,flagellar band.Then the fluorescence labeling of tubulin becomes more and more stronger as the flagella form in the spermatids.Based on the expression features of centrin and tubulin,it can be inferred that centrin is one of the organizer of the motile apparatuses,but not to be the components of themotile apparatuses.The main function of centrin may take part in formation of the centrioles,basal bodies and flagella of the spermatids.