Burgoyne R D
Physiological Laboratory, University of Liverpool, UK.
Pflugers Arch. 1995 Jun;430(2):213-9. doi: 10.1007/BF00374652.
The kinetics of exocytosis and membrane retrieval (endocytosis) were examined in bovine chromaffin cells using membrane capacitance measurement during whole-cell recording. At early times after breakthrough to the whole-cell recording mode, depolarisation for 1 s resulted in a fast (600 vesicles per s) exocytotic response and efficient membrane retrieval with a time constant of 25 s. The ability to activate fast exocytosis and retrieval was lost during intracellular dialysis, with a time constant of 40 s. At later times, a slow exocytotic response could be elicited with no membrane retrieval following single depolarisations. The wash-out of the responses appeared to be due to a rapid loss of a portion of the Ca2+ current. Trains of depolarisation at late times after breakthrough could elicit a fast (time constant 4 s) retrieval. These data show that in addition to a previously studied slow Ca(2+)-independent retrieval mechanism, chromaffin cells also possess an efficient and rapid retrieval pathway coupled to exocytosis that can be activated following depolarisation. The fast endocytosis appears to have a higher threshold for activation than exocytosis, probably due to a higher Ca2+ requirement. Rapid membrane retrieval appears to occur via a clathrin-independent pathway in chromaffin cells.
在全细胞记录过程中,通过膜电容测量研究了牛嗜铬细胞中的胞吐作用动力学和膜回收(内吞作用)。在进入全细胞记录模式后的早期,1秒的去极化导致快速(每秒600个囊泡)的胞吐反应和有效的膜回收,时间常数为25秒。在细胞内透析过程中,激活快速胞吐作用和回收的能力丧失,时间常数为40秒。在后期,单次去极化后可引发缓慢的胞吐反应,且无膜回收。反应的消退似乎是由于一部分Ca2+电流的快速丧失。在突破后的后期,一系列去极化可引发快速(时间常数4秒)的回收。这些数据表明,除了先前研究的缓慢的非Ca(2+)依赖性回收机制外,嗜铬细胞还拥有一种与胞吐作用相关的高效快速回收途径,该途径可在去极化后被激活。快速内吞作用的激活阈值似乎比胞吐作用更高,这可能是由于对Ca2+的需求更高。在嗜铬细胞中,快速的膜回收似乎是通过一种不依赖网格蛋白的途径发生的。