Yang Xia, Liu Ran, Brookes Simon J H
Qilu Hospital, Shandong University, Jinan 250012, China.
Sheng Li Xue Bao. 2006 Apr 25;58(2):171-6.
Intraganglionic laminar endings (IGLEs) have been supposed to be the mechanoreceptors in the gut by electrophysiological recording techniques. But the specialized morphology of IGLEs could not be displayed closely associated with this function and the mechanism that IGLEs act as the mechanotransduction sites in the gut is not yet well understood. In the present study, we used styryl dye FM1-43 combined with stretch stimulation in the guinea pig esophagus to test whether IGLEs acted as the mechano-sensitive receptors of the vagal afferent nerves. At the same time, the special structure of IGLEs displayed by FM1-43 was further confirmed by neurobiotin anterograde labeling technique. To further investigate the characteristics of IGLEs as mechanosensitive receptors, different drugs were used to block or stimulate IGLEs activation. Our results indicated that only in the stretched preparation could FM1-43 enter the IGLEs and completely display their specialized structure, which was consistent with that shown by neurobiotin. The amount of IGLEs shown by stretch-evoked FM1-43 staining was much more than that shown without stretch stimulation [(90.4 +/- 9.5) % vs (10.7 +/- 2.1) %, P<0.05]. Ca(2+), TTX (0.6 mumol/L), atropine (0.6 mumol/L), SKF (50 mumol/L), and gadolium (100 mumol/L) had no effect on the IGLEs activation. But for benzamil (100 mumol/L), an epithelial sodium channel blocker, activation of IGLEs by stretch stimulation was significantly blocked. The potent ATP analogue, alpha,beta-methylene ATP (100 mumol/L) could not activate FM1-43 staining without stretch. These results indicate that IGLEs are sensitive to mechanical stimulation. This could lead to the deduction that IGLEs act as the mechanoreceptors of vagal afferent nerve. IGLEs could transmit mechanical stimuli directly through ion channels, independent of neurotransmitter release and action potential propagation. The stretch-sensitive channels on IGLEs probably belong to the epithelial sodium channel family rather than voltage-gated sodium ion channels. Furthermore, styryl dye FM1-43 is a useful activity-dependent marker to demonstrate the structure and function of IGLEs in guinea pig esophagus.
通过电生理记录技术,人们一直认为神经节内分层终末(IGLEs)是肠道中的机械感受器。但IGLEs的特殊形态与该功能并未紧密相关,且IGLEs作为肠道机械转导位点的机制尚未完全清楚。在本研究中,我们将苯乙烯基染料FM1-43与豚鼠食管的牵张刺激相结合,以测试IGLEs是否作为迷走神经传入神经的机械敏感受体。同时,通过神经生物素顺行标记技术进一步证实了FM1-43所显示的IGLEs的特殊结构。为了进一步研究IGLEs作为机械敏感受体的特性,使用了不同的药物来阻断或刺激IGLEs的激活。我们的结果表明,只有在牵张的标本中,FM1-43才能进入IGLEs并完整显示其特殊结构,这与神经生物素所显示的一致。牵张诱发的FM1-43染色所显示的IGLEs数量远多于无牵张刺激时所显示的数量[(90.4±9.5)%对(10.7±2.1)%,P<0.05]。Ca(2+)、TTX(0.6 μmol/L)、阿托品(0.6 μmol/L)、SKF(50 μmol/L)和钆(100 μmol/L)对IGLEs的激活无影响。但对于上皮钠通道阻滞剂苯扎明(100 μmol/L),牵张刺激引起的IGLEs激活被显著阻断。强效ATP类似物α,β-亚甲基ATP(100 μmol/L)在无牵张时不能激活FM1-43染色。这些结果表明IGLEs对机械刺激敏感。由此可以推断,IGLEs作为迷走神经传入神经的机械感受器。IGLEs可以直接通过离子通道传递机械刺激,而不依赖于神经递质释放和动作电位传播。IGLEs上的牵张敏感通道可能属于上皮钠通道家族,而非电压门控钠离子通道。此外,苯乙烯基染料FM1-43是一种有用的活性依赖标记物,可用于展示豚鼠食管中IGLEs的结构和功能。