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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Calcium and tip growth in Neurospora crassa
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Calcium and tip growth in Neurospora crassa

机译：景天孢子中钙和尖端的生长

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We examined the ionic regulation of tip growth in Neurospora crassa by a combination of electrophysiology and confocal microscopy. To determine if transmembrane ionic fluxes are required for tip growth, we voltage clamped the membrane from -200 to +50 mV. In this voltage range, transmembrane ionic duxes would either reverse (e.g., K+) or change dramatically (e.g., Ca2+ influx) but had no effect on hyphal growth rates. Therefore, ionic fluxes (including Ca2+ influx) may not be required for tip growth. However, intracellular Ca2+ may still play an obligatory role in tip growth. To assess this possibility, we first increased cytosolic Ca2+ directly by ionophoresis. Elevated Ca2+ induced subapical branch initiation, often multiple tips. At hyphal tips, fluorescence ratio imaging using fluo-3 and fura-red revealed a pronounced tip-high Ca2+ gradient within 10 mu m of the tip in growing hyphae which was not observed in nongrowing hyphae. Injection of the Ca2+ chelator 1,2-bis(ortho-aminophenoxy)ethane-N,N,N',N'-tetrapotassium acetate consistently inhibited growth concomitantly with a depletion of intracellular Ca2+ and dissipation of the tip-high gradient. We conclude that Ca2+ plays a regulatory role in tip initiation and the maintenance of tip growth. Because plasma membrane ionic fluxes do not play a role in tip growth, we suggest that the tip-high Ca2+ gradient is generated from intracellular Ca2+ stores in the ascomycete N. crassa. [References: 78]
机译：我们通过电生理学和共聚焦显微镜的结合检查了神经孢子尖端生长的离子调节。为了确定尖端生长是否需要跨膜离子通量，我们将膜电压从-200钳位到+50 mV。在此电压范围内，跨膜离子二倍体会反转（例如，K +）或发生剧烈变化（例如，Ca2 +流入），但对菌丝的生长速率没有影响。因此，尖端生长可能不需要离子通量（包括Ca2 +流入）。但是，细胞内Ca2 +可能仍在尖端生长中起强制性作用。为了评估这种可能性，我们首先通过离子渗透法直接增加了胞质Ca2 +。 Ca2 +升高引起的根尖下分支开始，通常有多个尖端。在菌丝尖端，使用fluo-3和呋喃红的荧光比率成像显示，在生长的菌丝中，在菌丝的尖端10μm内有明显的尖端-高Ca2 +梯度，而在不生长的菌丝中未观察到。注入Ca2 +螯合剂1,2-双（邻氨基苯氧基）乙烷-N，N，N'，N'-乙酸四钾持续抑制细胞生长中的Ca2 +消耗并消除尖端高梯度。我们得出的结论是，Ca2 +在尖端引发和维持尖端生长中起调节作用。因为质膜离子通量在尖端生长中不起作用，所以我们建议从子​​囊猪笼草的细胞内Ca2 +储存产生尖端高的Ca2 +梯度。 [参考：78]

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