Gosain Hiteshika, Busch Karin B
Institute of Integrative Cell Biology and Physiology, Department of Biology, University of Muenster, Münster, North-Rhine-Westphalia, Germany.
Institute of Integrative Cell Biology and Physiology, Department of Biology, University of Muenster, Münster, North-Rhine-Westphalia, Germany.
Biophys Rep (N Y). 2024 Dec 11;4(4):100186. doi: 10.1016/j.bpr.2024.100186. Epub 2024 Nov 7.
The exocyst is an octameric protein complex that acts as a tether for GOLGI-derived vesicles at the plasma membrane during exocytosis. It is involved in membrane expansion during axonal outgrowth. Exo70 is a major subunit of the exocyst complex and is controlled by TC10, a Rho family GTPase. How TC10 affects the dynamics of Exo70 at the plasma membrane is not well understood. There is also evidence that TC10 controls Exo70 dynamics differently in nonpolar cells and axons. To address this, we used super-resolution microscopy to study the spatially resolved effects of TC10 on Exo70 dynamics in HeLa cells and the growth cone of cortical and hippocampal neurons. We generated single-particle localization and trajectory maps and extracted mean square displacements, diffusion coefficients, and alpha coefficients to characterize Exo70 diffusion. We found that the diffusivity of Exo70 was different in nonpolar cells and the growth cone of neurons. TC10 stimulated the mobility of Exo70 in HeLa cells but decreased the diffusion of Exo70 in the growth cone of cortical neurons. In contrast to cortical neurons, TC10 overexpression did not affect the mobility of Exo70 in the axonal growth cone of hippocampal neurons. These data suggest that mainly exocyst tethering in cortical neurons was under the control of TC10.
外泌体复合体是一种八聚体蛋白复合体,在胞吐过程中作为源自高尔基体的囊泡在质膜处的系链。它参与轴突生长过程中的膜扩张。Exo70是外泌体复合体的一个主要亚基,受Rho家族GTP酶TC10的调控。目前尚不清楚TC10如何影响Exo70在质膜上的动态变化。也有证据表明,TC10在非极性细胞和轴突中对Exo70动态变化的调控方式不同。为了解决这个问题,我们使用超分辨率显微镜研究了TC10对HeLa细胞以及皮质和海马神经元生长锥中Exo70动态变化的空间分辨效应。我们生成了单粒子定位和轨迹图,并提取了均方位移、扩散系数和α系数来表征Exo70的扩散。我们发现,Exo70在非极性细胞和神经元生长锥中的扩散率不同。TC10刺激了HeLa细胞中Exo70的移动性,但降低了皮质神经元生长锥中Exo70的扩散。与皮质神经元不同,TC10的过表达不影响海马神经元轴突生长锥中Exo70的移动性。这些数据表明,皮质神经元中的外泌体系链主要受TC10的控制。