Nishihara E, Shimmen T, Sonobe S
Department of Life Science, Graduate School of Life Science, University of Hyogo, Harima Science Park City, Hyogo, Japan.
Protoplasma. 2007;231(1-2):25-30. doi: 10.1007/s00709-007-0247-x. Epub 2007 Jul 3.
The contractile vacuole (CV) cycle of Amoeba proteus has been studied by phase contrast and electron microscopy. However, the understanding of membrane dynamics in this cycle is still poor. In this study, we used live imaging by fluorescence microscopy to obtain new insights. We succeeded in staining the CV with a styryl dye, FM 4-64 (N-(3-triethylammoniumpropyl)-4-(6-(4-(diethylamino)phenyl)hexatrienyl)pyridinium dibromide), and obtained the following results. (1) The CV membrane was directly stained with the dye in the external medium when the CV pore opened upon contraction. This indicates that transfer of plasma membrane to the CV does not occur. (2) The membrane dynamics during the CV cycle were elucidated. In particular, the fluorescent CV membrane was maintained as an aggregate just after contraction and the vacuole re-formed from the aggregate. Staining was maintained during continued contraction cycles. We conclude that the CV membrane is maintained during the CV cycle.
通过相差显微镜和电子显微镜对变形虫的收缩泡（CV）循环进行了研究。然而，对该循环中膜动力学的理解仍然不足。在本研究中，我们使用荧光显微镜进行实时成像以获得新的见解。我们成功地用一种苯乙烯基染料FM 4 - 64（N -（3 - 三乙铵丙基）- 4 -（6 -（4 -（二乙氨基）苯基）己三烯基）吡啶二溴化物）对收缩泡进行了染色，并获得了以下结果。（1）当收缩泡在收缩时其孔打开时，收缩泡膜在外部介质中直接被该染料染色。这表明质膜不会转移到收缩泡。（2）阐明了收缩泡循环期间的膜动力学。特别是，荧光收缩泡膜在收缩后刚形成聚集体，并从聚集体中重新形成液泡。在持续的收缩循环中染色得以维持。我们得出结论，收缩泡膜在收缩泡循环期间得以维持。
