Association of actin with sperm centrioles: isolation of centriolar complexes and immunofluorescent localization of actin.
Association of actin with sperm centrioles: isolation of centriolar complexes and immunofluorescent localization of actin.
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DOI：
10.1083/jcb.86.1.87
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发表时间：
1980-07
期刊：
影响因子：
--
通讯作者：
Clark WH Jr
中科院分区：
文献类型：
--
作者：
Kleve MG;Clark WH Jr
刺胞动物精子的中心粒在精子发生过程中与纹状特化（中心粒周围突起）相关。这些结构的作用被认为有三种功能：（a）精子鞭毛的锚定机制，（B）精子头部和尾部之间通讯的信号传递机制，以及（c）精子鞭毛运动功能中的收缩机制。为了研究这些建议的功能，我们开发了一种技术，用于分离和纯化的Hydracactinia精子远端中心体与附加的pericentriolar进程。整个精子和pericentriolar过程蛋白的SDS聚丙烯酰胺电泳图谱显示一个突出的蛋白质，comigrates与兔和对虾肌肉肌动蛋白。为了标记和定位水螅体精子中的肌动蛋白，我们在兔中制备了一种高度特异性的抗无脊椎动物肌动蛋白的抗血清，该抗血清与无脊椎动物和脊椎动物的肌肉和非肌肉肌动蛋白发生交叉反应。免疫荧光双抗体标记的水螅精子与抗肌动蛋白已证明肌动蛋白的存在下，在pericentriolar过程区的精子。在早期的报道中，已经提出，如果中心粒周围突起收缩，则可以改变水螅精子的中段不对称性，从而促进这些细胞对卵释放的化学引诱物的反应所表现出的定向运动。目前的结果支持这一假设。
The centrioles of cnidarian sperm associate with striated specializations (pericentriolar processes) during spermiogenesis. Three functions have been proposed for the role of these structures: (a) an anchoring mechanism for the sperm flagellum, (b) a signal-transmitting mechanism for communication between sperm head and tal, and (c) a contractile mechanism involved in motor function of the sperm flagellum. To investigate these proposed functions, we developed a technique for the isolation and purification of Hydractinia sperm distal centriles with attached pericentriolar processes. SDS polyacrylamide electrophoretic profiles of whole sperm and pericentriolar process proteins revealed a prominent protein that comigrates with rabbit and penaeid shrimp muscle actin. To label and localize actin in hydroid spem, we produced in rabbits a highly specific antiserum to invertebrate actin that cross-reacts with both invertebrate and vertebrate muscle and nonmuscle actin. Immunofluorescent double antibody labeling of hydroid sperm with antiactin has demonstrated the presence of actin in the pericentriolar process region of the sperm. In earlier reports, it has been proposed that pericentriolar processes, if contractile, could alter the mid- piece asymmetry of hydroid sperm, facilitating the directional motility that these cells demonstrate in respone to egg-released chemoattractants. The present results support this hypothesis.