Iwadate Yoshiaki, Suzaki Toshinobu
Department of Life Science, Faculty of Integrated Arts and Sciences, The University of Tokushima, Tokushima, Japan.
Cell Motil Cytoskeleton. 2004 Apr;57(4):197-206. doi: 10.1002/cm.10165.
Internal Ca2+ levels control the pattern of ciliary and flagellar beating in eukaryotes. In ciliates, ciliary reversal is induced by a rise in intra-ciliary Ca2+, but the mechanism by which Ca2+ induces reversal is not known. We injected the fluorescent Ca2+ indicator Calcium Green into a ciliate Didinium nasutum and observed the intra-ciliary Ca2+ level during the initial reversed stroke preceding spontaneous cyclic reversed beating. In D. nasutum, Ca2+ rose throughout the length of the cilia undergoing initial reversed stroke. Electron microscopy with a combined oxalate-pyroantimonate method showed Ca2+ deposits distributed throughout the reversed cilia. We injected caged Ca2+ into D. nasutum and irradiated the base or mid region of the cilia with UV to locally increase Ca2+ concentration. Uncaging Ca2+ in the middle of the cilia produced reversal distally, but not proximally to the site of Ca2+ release. These results strongly suggest that not only Ca2+ influx sites, but also Ca2+ binding sites and vectoral bending machineries for ciliary reversal, are distributed throughout the cilium.
细胞内钙离子水平控制真核生物中纤毛和鞭毛的摆动模式。在纤毛虫中,纤毛内钙离子浓度升高会诱导纤毛反转,但钙离子诱导反转的机制尚不清楚。我们将荧光钙离子指示剂钙黄绿素注入一种纤毛虫——多膜大草履虫中,并在自发周期性反转摆动之前的初始反转冲程期间观察纤毛内的钙离子水平。在多膜大草履虫中,在经历初始反转冲程的整个纤毛长度上钙离子浓度都升高。采用草酸盐 - 焦锑酸盐联合方法的电子显微镜观察显示,钙离子沉积物分布在整个反转的纤毛中。我们将笼锁钙离子注入多膜大草履虫,并用紫外线照射纤毛的基部或中部区域以局部增加钙离子浓度。在纤毛中部释放笼锁钙离子会在远端产生反转,但在钙离子释放位点的近端则不会。这些结果有力地表明,不仅钙离子流入位点,而且钙离子结合位点以及用于纤毛反转的矢量弯曲机制都分布在整个纤毛中。