Middleton P, Jaramillo F, Schuetze S M
Proc Natl Acad Sci U S A. 1986 Jul;83(13):4967-71. doi: 10.1073/pnas.83.13.4967.
We have studied the function of acetylcholine (AcCho) receptors (AcChoRs) in rat soleus endplates before and after exposing the muscles to forskolin, a potent activator of adenylate cyclase. AcChoR function was tested by recording the membrane depolarization evoked by pulses of ionophoretically applied AcCho. Brief (2 msec) AcCho pulses delivered at 7 Hz evoked constant responses at untreated endplates. In contrast, after 10-100 microM forskolin was added to the bath, responses to similar pulse trains fell by as much as 80% within 1 sec. AcCho sensitivity recovered completely in less than 1 min after the pulses were stopped but fell again when the pulses were resumed. Similarly, longer (1 sec) ionophoretic AcCho pulses evoked roughly constant responses at control endplates, but after forskolin treatment the depolarization fell by one-half within less than 200 msec. These results indicate that forskolin increases the rate at which AcChoRs desensitize when exposed to agonist. Focal extracellular recordings showed that 20-100 microM forskolin also increased the decay rate of miniature endplate currents, indicating that forskolin may decrease AcChoR channel open time. Inhibitors of cAMP phosphodiesterase increased the potency of forskolin. When used alone, these inhibitors had effects similar to those of forskolin but smaller. Patch-clamp experiments indicated that forskolin at 100 microM may also interact with AcChoR channels directly, but at 20 microM this effect is negligible. Therefore, it is likely that the forskolin effects were mediated primarily by increased levels of intracellular cAMP.
我们研究了将大鼠比目鱼肌终板暴露于强效腺苷酸环化酶激活剂福斯高林前后,乙酰胆碱(AcCho)受体(AcChoRs)的功能。通过记录离子电泳施加的AcCho脉冲诱发的膜去极化来测试AcChoR功能。在未经处理的终板上,以7 Hz频率施加的短暂(2毫秒)AcCho脉冲可诱发恒定反应。相比之下,向浴液中加入10 - 100 microM福斯高林后,对类似脉冲序列的反应在1秒内下降多达80%。脉冲停止后不到1分钟,AcCho敏感性完全恢复,但脉冲恢复时又再次下降。同样,较长(1秒)的离子电泳AcCho脉冲在对照终板上诱发大致恒定的反应,但福斯高林处理后,去极化在不到200毫秒内下降了一半。这些结果表明,福斯高林增加了AcChoRs在暴露于激动剂时脱敏的速率。局部细胞外记录显示,20 - 100 microM福斯高林也增加了微小终板电流的衰减速率,表明福斯高林可能会减少AcChoR通道开放时间。环磷酸腺苷(cAMP)磷酸二酯酶抑制剂提高了福斯高林的效力。单独使用时,这些抑制剂的作用与福斯高林相似,但程度较小。膜片钳实验表明,100 microM的福斯高林也可能直接与AcChoR通道相互作用,但20 microM时这种作用可忽略不计。因此,福斯高林的作用可能主要是由细胞内cAMP水平升高介导的。