Albillos Almudena, Gil Amparo, González-Vélez Virginia, Pérez-Álvarez Alberto, Segura Javier, Hernández-Vivanco Alicia, Caba-González José Carlos
Departamento de Farmacología y Terapéutica, Facultad de Medicina, Universidad Autónoma de Madrid, 28029 Madrid, Spain.
J Comput Neurosci. 2013 Feb;34(1):27-37. doi: 10.1007/s10827-012-0404-x. Epub 2012 Jun 13.
Chromaffin cells have been widely used to study neurosecretion since they exhibit similar calcium dependence of several exocytotic steps as synaptic terminals do, but having the enormous advantage of being neither as small or fast as neurons, nor as slow as endocrine cells. In the present study, secretion associated to experimental measurements of the exocytotic dynamics in human chromaffin cells of the adrenal gland was simulated by using a model that combines stochastic and deterministic approaches for short and longer depolarizing pulses, respectively. Experimental data were recorded from human chromaffin cells, obtained from healthy organ donors, using the perforated patch configuration of the patch-clamp technique. We have found that in human chromaffin cells, secretion would be mainly managed by small pools of non-equally fusion competent vesicles, slowly refilled over time. Fast secretion evoked by brief pulses can be predicted only when 75% of one of these pools (the "ready releasable pool" of vesicles, abbreviated as RRP) are co-localized to Ca²⁺ channels, indicating an immediately releasable pool in the range reported for isolated cells of bovine and rat (Álvarez and Marengo, J Neurochem 116:155-163, 2011). The need for spatial correlation and close proximity of vesicles to Ca²⁺ channels suggests that in human chromaffin cells there is a tight control of those releasable vesicles available for fast secretion.
嗜铬细胞已被广泛用于研究神经分泌,因为它们在几个胞吐步骤中表现出与突触终末相似的钙依赖性,但具有巨大优势,既不像神经元那么小且快速,也不像内分泌细胞那么缓慢。在本研究中,通过使用一种分别针对短和长去极化脉冲结合随机和确定性方法的模型,模拟了与肾上腺人类嗜铬细胞胞吐动力学实验测量相关的分泌。使用膜片钳技术的穿孔膜片配置,从健康器官捐献者获取的人类嗜铬细胞记录实验数据。我们发现,在人类嗜铬细胞中,分泌主要由少量融合能力不等的囊泡池控制,这些囊泡池随时间缓慢重新填充。只有当这些池中的一个池(囊泡的“可立即释放池”,简称为RRP)的75%与Ca²⁺通道共定位时,才能预测由短脉冲诱发的快速分泌,这表明在牛和大鼠分离细胞报道的范围内存在一个可立即释放池(阿尔瓦雷斯和马伦戈,《神经化学杂志》116:155 - 163,2011)。囊泡与Ca²⁺通道需要空间相关性和紧密接近性,这表明在人类嗜铬细胞中,对那些可用于快速分泌的可释放囊泡有严格控制。