Grider J R, Bonilla O M
Department of Physiology, Medical College of Virginia, Richmond 23298.
Am J Physiol. 1994 Aug;267(2 Pt 1):G322-7. doi: 10.1152/ajpgi.1994.267.2.G322.
The expression of three enteric neuropeptides was examined in freshly dispersed ganglia and in ganglia cultured for up to 28 days. During culture, glial cells grew into a flat sheet surrounding a cluster of neurons identified with neuron-specific enolase (13 +/- 2/ganglion), which remained constant throughout the period of culture. The neurons underwent a distinctive temporal change, resulting in overexpression of substance P (SP), normal expression of somatostatin, and virtual suppression of vasoactive intestinal peptide (VIP). Three weeks after the start of culture, the ganglia contained and released (in response to 55 nM KCl, 0.1 mM 1,1-dimethyl-4-phenylpiperazinium, or 1 microM gastrin-releasing peptide) twice as much SP as freshly dispersed ganglia, corresponding to a sevenfold increase per cultured neuron; content and release of somatostatin did not change. SP content and release declined to 1.5% of those found in control cultures when nonneuronal cells were suppressed with cytosine arabinoside but were partially restored (13-17% of control) by nerve growth factor. In marked contrast, VIP was minimally (< 1%) present in and released from ganglia after the third day in culture. Suppression of VIP could reflect a selective loss of VIP neurons and/or VIP expression.
在新鲜分散的神经节以及培养长达28天的神经节中检测了三种肠神经肽的表达。在培养过程中,神经胶质细胞生长成围绕一群用神经元特异性烯醇化酶鉴定的神经元(13±2/神经节)的扁平薄片,在整个培养期间保持恒定。神经元经历了独特的时间变化,导致P物质(SP)过表达、生长抑素正常表达以及血管活性肠肽(VIP)几乎完全被抑制。培养开始三周后,神经节中所含的SP(响应于55 nM氯化钾、0.1 mM 1,1-二甲基-4-苯基哌嗪或1 μM胃泌素释放肽)释放量是新鲜分散神经节的两倍,相当于每个培养神经元增加了七倍;生长抑素的含量和释放没有变化。当用阿糖胞苷抑制非神经元细胞时,SP的含量和释放降至对照培养物中的1.5%,但神经生长因子可使其部分恢复(对照的13 - 17%)。与之形成鲜明对比的是,培养第三天后,神经节中VIP的含量和释放量极少(<1%)。VIP的抑制可能反映了VIP神经元的选择性丧失和/或VIP表达的丧失。