Dinkelacker V, Voets T, Neher E, Moser T
Department of Membrane Biophysics, Max-Planck-Institute for Biophysical Chemistry, D-37077 Göttingen, Germany.
J Neurosci. 2000 Nov 15;20(22):8377-83. doi: 10.1523/JNEUROSCI.20-22-08377.2000.
Maturation of exocytic vesicles to the release-ready state is regulated by several factors, including intracellular calcium concentration (Ca(2+)) and the state of protein phosphorylation. Here we investigated the effects of temperature on the recovery from depletion of the readily releasable pool (RRP) of vesicles in adrenal chromaffin cells. Exocytosis and Ca(2+) were monitored by combined membrane capacitance and fura-2 measurements. At higher temperatures, a faster pool refilling and a larger RRP size were observed. The time constants of the recovery from depletion ranged from 3.6 to 1.1 sec (22 and 37 degrees C, respectively) yielding a Q(10) of 2.3. The changes of the Ca(2+) signal between the different temperatures could not account for the differences in recovery kinetics. At 32 and 37 degrees C, we observed a transient overfilling of the RRP after pool depletion, which stands in clear contrast to the sustained secretory depression seen at lower temperatures. The overshoot in RRP size was very prominent in cells with lower basal Ca(2+), hence with a large difference between prestimulus and poststimulus Ca(2+). In cells with higher basal Ca(2+), the pool was larger under steady-state conditions but showed less overfilling on stimulation. We conclude that vesicle maturation is markedly accelerated at physiological temperature, thus allowing for a rapid adaptation of the pool size to the relatively short-lived Ca(2+) transient.
分泌囊泡成熟至可释放状态受多种因素调控,包括细胞内钙浓度(Ca(2+))和蛋白质磷酸化状态。在此,我们研究了温度对肾上腺嗜铬细胞中囊泡易释放池(RRP)耗竭后恢复的影响。通过联合膜电容和fura-2测量来监测胞吐作用和Ca(2+)。在较高温度下,观察到池再填充更快且RRP尺寸更大。耗竭后恢复的时间常数范围为3.6至1.1秒(分别为22和37摄氏度),Q(10)为2.3。不同温度之间Ca(2+)信号的变化无法解释恢复动力学的差异。在32和37摄氏度时,我们观察到池耗竭后RRP出现短暂的过度填充,这与在较低温度下看到的持续性分泌抑制形成鲜明对比。RRP尺寸的过冲在基础Ca(2+)较低的细胞中非常显著,因此刺激前和刺激后Ca(2+)之间存在较大差异。在基础Ca(2+)较高的细胞中,池在稳态条件下更大,但刺激时过度填充较少。我们得出结论,在生理温度下囊泡成熟明显加速,从而使池大小能够快速适应相对短暂的Ca(2+)瞬变。