Mechanisms underlying coordinated regulation of exo- and endocytosis in BY-2 cells

膜片钳电容测量允许在活细胞的质膜上的单个融合和裂变事件的动力学和大小的直接分析。这项技术为揭示外吞作用和内吞作用的机制提供了一个有吸引力的工具。为了研究植物细胞外吞作用和内吞作用的潜在调节因子，我们将使用BY-2细胞培养原生质体的电容测量作为模型系统。我们的研究重点将集中在囊泡融合和裂变的当前主题，如细胞对渗透应激的反应，网格蛋白介导的内吞作用和磷酸肌苷在外胞吞作用中的作用。应用特异性抑制剂和基因干扰内吞途径和磷酸肌苷合成将用于研究潜在的分子机制。此外，我们将建立使用花粉管进行膜片钳电容测量，为研究生理背景下的单融合事件提供新的工具。这不仅将增加我们对花粉管分泌的认识，而且还将提供有关胞吐作用时空调节机制的信息。

Patch-clamp capacitance measurements allow the direct analysis of the kinetic and size of single fusion and fission events at the plasma membrane of living cells. This technique provides an attractive tool to unravel the mechanisms of exo- and endocytosis. To study the role of potential regulators of exo- and endocytosis in living plant cells we will therefore use capacitance measurements on protoplasts from BY-2 cell cultures as a model system. The focus of our investigations will be on current topics of vesicle fusion and fission such as the response of cells to osmotic stress, the role of clathrin mediated endocytosis and the involvement of phosphoinositides in exoand endocytosis. Application of specific inhibitors and genetic interference with the endocytotic pathway and phosphoinositide synthesis will be used to study the underlying molecular mechanisms. In addition, we will establish the use of pollen tubes for patch-clamp capacitance measurements to provide a new tool for the investigating of single fusion events in a physiological context. This will not only increase our knowledge of secretion in pollen tubes but also provide information about the mechanisms involved in spatial and temporal regulation of exocytosis in general.