Armisén R, Sierralta J, Vélez P, Naranjo D, Suárez-Isla B A
Departamento de Fisiología y Biofísica, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
Am J Physiol. 1996 Jul;271(1 Pt 1):C144-53. doi: 10.1152/ajpcell.1996.271.1.C144.
The bursting behavior of ryanodine-sensitive single Ca2+ release channels present in chicken cerebellum endoplasmic reticulum (ER), rat hippocampus ER, and frog and rabbit skeletal muscle sarcoplasmic reticulum was established. Unconditional dwell time distributions fitted by the maximum likelihood method reveal at least three open and closed exponential components. Trains of low open probability (P(o)) bursts were interspersed with trains of high P(o) bursts (> or = 0.8) in all the ryanodine receptor isotypes tested. The gating kinetics of the Ca2+ release channels were defined in long recordings by analyzing burst sequences and gamma distributions of average intraburst open (T(o)) and closed times (Tc). The gamma distributions of T(o) had two gamma components, suggesting the existence of two distinct burst types. In contrast, the gamma distributions of Tc had only one component. The correlation between consecutive burst pairs was defined in terms of T(o) and then statistically tested by 2 x 2 matrix contingency analysis. The probability that the ubiquitous sequential burst pattern was generated by random occurrence was < 0.01 (two-tailed Fisher's exact test). Temporal correlations were observed in all ryanodine receptor isotypes under a variety of experimental conditions. These data strongly suggest that single Ca2+ release channels switch slowly between modes of gating. We propose that the effects of agonists of Ca2+ release channels such as Ca2+ itself can be explained as concentration-dependent changes in the availability of each mode.
已确定存在于鸡小脑内质网(ER)、大鼠海马体ER以及青蛙和兔子骨骼肌肌浆网中的兰尼碱敏感型单个Ca2+释放通道的爆发行为。通过最大似然法拟合的无条件停留时间分布揭示了至少三个开放和关闭的指数成分。在所有测试的兰尼碱受体同种型中,低开放概率(P(o))爆发序列与高P(o)爆发序列(≥0.8)交替出现。通过分析爆发序列以及平均爆发内开放时间(T(o))和关闭时间(Tc)的伽马分布,在长时间记录中确定了Ca2+释放通道的门控动力学。T(o)的伽马分布有两个伽马成分,表明存在两种不同的爆发类型。相比之下,Tc的伽马分布只有一个成分。根据T(o)定义连续爆发对之间的相关性,然后通过2×2矩阵列联分析进行统计学检验。由随机发生产生普遍存在的连续爆发模式的概率<0.01(双尾费舍尔精确检验)。在各种实验条件下,在所有兰尼碱受体同种型中均观察到时间相关性。这些数据强烈表明单个Ca2+释放通道在门控模式之间缓慢切换。我们提出,Ca2+释放通道激动剂(如Ca2+本身)的作用可以解释为每种模式可用性的浓度依赖性变化。