Schulzke J D, Fromm M, Hegel U, Riecken E O
Medizinische Klinik, Abteilung für Gastroenterologie, Freie Universität Berlin.
Pflugers Arch. 1989 Jun;414(2):216-21. doi: 10.1007/BF00580966.
The initial phase of in vitro experiments in Ussing-type chambers on large intestine is characterized by short-circuit currents (ISC) declining from high starting values to a lower plateau within 0.5 h. The origin of this "initial ISC-transient" was investigated by ISC measurements on partially stripped segments of rat rectal colon. Transport was pre-stimulated in vivo by keeping animals in barbiturate-anesthesia for 5 h prior to tissue preparation. This procedure caused by endogenous aldosterone-liberation amiloride-sensitive Na-absorption to become the predominant electrogenic transport. The initial ISC-transient was abolished by tetrodotoxin (TTX, 1 microM), indicating a neuronal mediation of this phenomenon. In order to identify the transport which was subject to neuronal control, the amiloride-sensitive Na-absorption was measured during electrical field stimulation (bipolar rectangular pulses: 5 Hz, 1 ms, +/- 6 mA). There was no difference to unstimulated controls. In contrast, the initial ISC-transient was dependent on Cl in the bath following Michaelis-Menten-kinetics (KM = 20 mM) and could be prevented by 10 microM serosal bumetanide. Then, initial filling of the Ussing-chamber was imitated during the course of the experiment by removal and immediate re-addition of the bathing fluid. This procedure caused ISC-changes of similar appearance as the initial ISC-transient. To verify that indeed mechanical stretch is the sensory stimulus triggering the initial ISC-transient, the effect of small pressure oscillations was studied. This also produced an ISC-transient which was TTX-sensitive and was abolished after removal of the submucosal plexus Meissner by total stripping.(ABSTRACT TRUNCATED AT 250 WORDS)
在乌斯辛型小室中对大肠进行的体外实验初始阶段的特征是,短路电流(ISC)在0.5小时内从高起始值下降到较低的平稳期。通过对大鼠直肠结肠部分剥离段进行ISC测量,研究了这种“初始ISC瞬变”的起源。在组织制备前,通过将动物置于巴比妥类麻醉下5小时,在体内预先刺激转运。该程序通过内源性醛固酮释放导致对阿米洛利敏感的钠吸收成为主要的电转运。河豚毒素(TTX,1 microM)消除了初始ISC瞬变,表明该现象由神经元介导。为了确定受神经元控制的转运,在电场刺激(双极矩形脉冲:5 Hz,1 ms,+/- 6 mA)期间测量了对阿米洛利敏感的钠吸收。与未刺激的对照没有差异。相反,初始ISC瞬变在浴液中依赖于氯离子,遵循米氏动力学(KM = 20 mM),并且可以被10 microM的浆膜布美他尼阻止。然后,在实验过程中通过去除并立即重新添加浴液来模拟乌斯辛小室的初始填充。该程序引起的ISC变化与初始ISC瞬变外观相似。为了验证机械拉伸确实是触发初始ISC瞬变的感觉刺激,研究了小压力振荡的影响。这也产生了一个对TTX敏感的ISC瞬变,在通过完全剥离去除黏膜下梅斯纳丛后消失。(摘要截断于250字)