Plattner H, Braun C, Klauke N, Länge S
Universität Konstanz, Fakultät für Biologie, Germany.
J Membr Biol. 1994 Nov;142(2):229-40. doi: 10.1007/BF00234945.
Paramecium tetraurelia wild-type (7S) cells respond to 2.5 mM veratridine by immediate trichocyst exocytosis, provided [Ca2+]o (extracellular Ca2+ concentration) is between about 10(-4) to 10(-3) M as in the culture medium. Exocytosis was analyzed by light scattering, light and electron microscopy following quenched-flow/freeze-fracture analysis. Defined time-dependent stages occurred, i.e., from focal (10 nm) membrane fusion to resealing, all within 1 sec. Veratridine triggers exocytosis also with deciliated 7S cells and with pawn mutants (without functional ciliary Ca channels). Both chelation of Ca2+o or increasing [Ca2+]o to 10(-2) M inhibit exocytotic membrane fusion. Veratridine does not release Ca2+ from isolated storage compartments and it is inefficient when microinjected. Substitution of Na+o for N-methylglucamine does not inhibit the trigger effect of veratridine which also cannot be mimicked by aconitine or batrachotoxin. We conclude that, in Paramecium cells, veratridine activates Ca channels (sensitive to high [Ca2+]o) in the somatic, i.e., nonciliary cell membrane and that a Ca2+ influx triggers exocytotic membrane fusion. The type of Ca channels involved remains to be established.
在细胞外钙离子浓度([Ca2+]o)处于大约10^(-4)至10^(-3)M之间(如在培养基中)时,四膜虫野生型(7S)细胞会通过立即释放刺丝泡来响应2.5 mM藜芦定。通过光散射、光镜和电镜观察,在淬灭流动/冷冻断裂分析后对胞吐作用进行了分析。确定了时间依赖性阶段,即从局部(10纳米)膜融合到重新封闭,整个过程在1秒内完成。藜芦定也能触发去纤毛7S细胞和pawn突变体(无功能性纤毛钙通道)的胞吐作用。细胞外钙离子螯合或细胞外钙离子浓度增加到10^(-2)M均会抑制胞吐性膜融合。藜芦定不会从分离的储存隔室中释放钙离子,微注射时效果不佳。用N-甲基葡糖胺替代细胞外钠离子不会抑制藜芦定的触发作用,乌头碱或蟾毒素也无法模拟这种作用。我们得出结论,在四膜虫细胞中,藜芦定激活体细胞(即非纤毛细胞膜)中的钙通道(对高细胞外钙离子浓度敏感),并且钙离子内流触发胞吐性膜融合。所涉及的钙通道类型仍有待确定。