Distribution and role of calmodulin in tip growing hyphae of Saprolegnia ferax*
Distribution and role of calmodulin in tip growing hyphae of Saprolegnia ferax*
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DOI：
10.1038/cr.1995.24
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发表时间：
1995-12
期刊：
影响因子：
44.1
通讯作者：
S. Yuan;Ling-Xiao Lu;S. Xue
中科院分区：
文献类型：
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作者：
S. Yuan;Ling-Xiao Lu;S. Xue
由于三氟拉嗪(TFP)直接染色生长的菌丝会导致水霉菌菌丝内原生质的迁移，因此开发了一种改进的固定生长菌丝的染色方法。当水霉菌丝用多聚甲醛预固定时，TFP荧光显示出清晰的尖高Ca 2+ 激活钙调蛋白（CaM）梯度。通过间接免疫组化染色，我们证明总CaM也主要集中在菌丝顶端。值得注意的是，当菌丝尖端生长停止时，高强度的TFP荧光通常在非生长尖端消失，而在新出现的生长尖端中变得明显，这表明CaM浓度的变化与菌丝中生长点的转移一致。 2× 10− 5 mol/L CaM抑制剂TFP可使菌丝尖端生长速度和生长菌丝尖端百分比降低至50%左右，并诱导菌丝尖端多分枝。这些发现强烈表明菌丝尖端中高浓度的钙调蛋白可能参与菌丝尖端生长的调节。
Since direct staining of growing hyphae with trifluoperazine (TFP) could result in the migration of protoplasm in hyphae of water mold Saprolegnia ferax, an improved staining method for fixation of growing hyphae was developed. When hyphae of Saprolegnia ferax were pre-fixed with paraformaldehyde, the TFP fluorescence showed a clear tip-high Ca 2+-activated calmodulin (CaM) gradient. By indirect immunohistochemical staining, we proved that total CaM also mainly concentrated in the apex of hyphae. It is noticeble that when the hyphal tip growth ceased, the high intensity of TFP fluorescence usually disappeared in non-growing tips and became evident in the newly emerged growing tips, which implied that the concentration of CaM shifted in concord with the transfer of growing point in hyphae. 2× 10− 5 mol/L CaM inhibitor TFP could reduce the tip growth rate and the percentage of growing hyphal tips to about 50%, and induce multibranching of hyphal tips. These findings strongly suggest that the high concentration of calmodulin in the hyphal tip may be involved in regulation of hyphal tip growth.