Perez C, Roco M, Castro A, Dupre E, Schatten G, Barros C
Laboratory of Embryology, Faculty of Biological Sciences, Pontifical Catholic University of Chile, Santiago.
Mol Reprod Dev. 1991 Apr;28(4):373-9. doi: 10.1002/mrd.1080280410.
Sperm from the decapod crustacean Rhynchocinetes typus undergo dramatic shape changes as they pass from the vas deferens to seawater and interact with the oocyte envelopes. Using FITC-phalloidin and antitubulin antibodies, we were able to localize microfilaments and a tubulin-like protein in R. typus spermatozoon. Microfilaments and the tubulin-like protein were associated with the sperm rays and spines, but were absent at the spike and at its base. Folded and unfolded spermatozoa display similar fluorescence patterns. SDS-PAGE of whole spermatozoa and electrotransfer to nitrocellulose confirmed the presence of actin and two proteins at 97 kd and 120 kd that bind to tubulin antibodies (tubulin-like proteins). These results demonstrate the presence of actin, but not tubulin, and localize microfilaments in these sperm. It is proposed that this cytoskeletal component is active in sperm during crustacean fertilization.
十足目甲壳动物罗氏沼虾(Rhynchocinetes typus)的精子在从输精管进入海水并与卵母细胞包膜相互作用的过程中会发生显著的形态变化。使用异硫氰酸荧光素标记的鬼笔环肽(FITC-phalloidin)和抗微管蛋白抗体,我们能够在罗氏沼虾精子中定位微丝和一种微管蛋白样蛋白。微丝和微管蛋白样蛋白与精子射线和刺相关,但在尖刺及其基部不存在。折叠和未折叠的精子显示出相似的荧光模式。全精子的十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)以及向硝酸纤维素膜的电转移证实了肌动蛋白和两种与微管蛋白抗体结合的97千道尔顿和120千道尔顿的蛋白质(微管蛋白样蛋白)的存在。这些结果证明了肌动蛋白而非微管蛋白的存在,并在这些精子中定位了微丝。有人提出,这种细胞骨架成分在甲壳动物受精过程中对精子起作用。