Stommel E W, Stephens R E, Alkon D L
J Cell Biol. 1980 Dec;87(3 Pt 1):652-62. doi: 10.1083/jcb.87.3.652.
We have investigated the role of motile cilia in mechanotransduction by statocysts of the nudibranch mollusk Hermissenda crassicornis. Movement of the cilia that experience the weight of statoconia causes increased variance of voltage noise and membrane depolarization of the statocyst hair cell. Two complementary approaches were used to immobilize the cilia. Vanadate anion was iontophoretically injected into hair cells. This reversible inhibitor of vibratile form and to assume a more classic, pliable beat pattern. Voltage noise decreased as the cilia slowed and bent more extremely, nearly disappearing as motility was lost. When the intracellular vanadate concentration approached 10(-5) M, the cilia were arrested in an effective stroke against the cell membrane. The cell no longer depolarized upon gravitational or local mechanical stimulation. Rapid reversal of ciliary inhibition by norepinephrine or slow reversal with time restored both the voltage noise and depolarization response. Cilia were rendered rigid and upright by covalent cross-linkage of their membrane "sleeve" to the 9 + 2 axoneme, using the photoactivated, lipophilic, bifunctional agent 4,4'-dithiobisphenyl azide. In the initial stages of cross-linkage, the cilia remained vibratile but slowed and moved through wider excursions. Voltage noise decreased in frequency but increased in amplitude. When the cilia were fully arrested, voltage noise was minimized while the resting potential and membrane resistance remained essentially constant. Mechanical stimulation of the rigid cilia, normal to the cell membrane, elicited a generator potential of the same amplitude but of greater duration than before treatment. Because cilia that are partially arrested by vanadate undergo increased bending, although the hair cell shows decreased noise, neither the axoneme nor the ciliary membrane proper would appear to be sites of direct transduction. In cells with beating but stiffened cilia, however, the voltage noise becomes amplified, implying an increased efficiency of transduction. We suggest that active but rigid flexure of the axoneme is involved in amplification and continuous signal detection. The basal insertion area is the most likely transduction site, being the terminal leverage point through which force is applied to the plasma membrane via the flexing ciliary shaft.
我们研究了能动纤毛在海蛞蝓(Hermissenda crassicornis)平衡囊机械转导中的作用。感受平衡石重量的纤毛运动导致电压噪声方差增加以及平衡囊毛细胞的膜去极化。我们使用了两种互补的方法来固定纤毛。将钒酸盐阴离子通过离子电泳注入毛细胞。这种振动形式的可逆抑制剂使纤毛呈现出更经典、更柔韧的摆动模式。随着纤毛变慢并更剧烈地弯曲,电压噪声降低,当失去运动能力时几乎消失。当细胞内钒酸盐浓度接近10^(-5) M时,纤毛在对细胞膜的有效摆动中停止。细胞在重力或局部机械刺激下不再去极化。去甲肾上腺素对纤毛抑制的快速逆转或随时间的缓慢逆转恢复了电压噪声和去极化反应。使用光活化的亲脂性双功能试剂4,4'-二硫代双苯叠氮,通过其膜“套筒”与9 + 2轴丝的共价交联使纤毛变得僵硬且直立。在交联的初始阶段,纤毛仍能摆动但变慢且摆动幅度更大。电压噪声频率降低但幅度增加。当纤毛完全停止时,电压噪声最小化,而静息电位和膜电阻基本保持不变。垂直于细胞膜对僵硬纤毛进行机械刺激,引发的发生器电位幅度与处理前相同,但持续时间更长。因为被钒酸盐部分阻止运动的纤毛弯曲增加,尽管毛细胞的噪声降低,但轴丝和纤毛膜本身似乎都不是直接转导的位点。然而,在纤毛摆动但变硬的细胞中,电压噪声会放大,这意味着转导效率提高。我们认为轴丝的主动但僵硬的弯曲参与了放大和连续信号检测。基部插入区域是最可能的转导位点,它是通过弯曲的纤毛轴将力施加到质膜的终端杠杆点。