Abstract

The crayfish stretch receptor consisting of the single mechanoreceptor neurons enveloped by satellite glial cells is the simplest functioning neuroglial preparation. However, during isolation, its axons are usually transected that eliminates afferent regulation and induces complex axotomy-related signaling responses in neurons and satellite glia. We developed new microsurgical method of crayfish stretch receptor isolation, which preserves connections of sensory neurons to the ventral nerve cord ganglion. The stretch receptor may either remain on the abdominal carapace, or be completely isolated. In both cases, it may be either intact, or axotomized. The integrity of axons was confirmed by firing recording from proximal and distal axon points. Normal, necrotic and apoptotic cells were visualized using double fluorochroming with Hoechst 33342 and propidium iodide. The isolated mechanoreceptor neurons maintain regular firing during 8–10 or more hours. Glial cells surrounding non-axotomized neurons demonstrate lower necrosis and apoptosis levels than the axotomized ones. Unlike the existing method, in which the sensory neurons were axotomized, the present method preserves links between the sensory neurons and the ganglion and makes possible to avoid consequences of axotomy in neurons and satellite glia. The present neuroglial preparation may be used as a simple but informative model object in studies of axotomy-induced degeneration and survival of peripheral neurons, the role of glia in neuron injury, the signaling mechanisms of neuroglial interactions, and the effects of diverse physical and chemical factors on neuronal and glial cells.

中文翻译：

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分离小龙虾腹部拉伸受体的方法，可维持感觉神经元与腹神经索神经节的连接。

抽象的

由小卫星神经胶质细胞包被的单个机械感受器神经元组成的小龙虾牵张受体是功能最简单的神经胶质制剂。然而，在分离过程中，其轴突通常被横断，从而消除传入调节，并在神经元和卫星神经胶质细胞中诱导复杂的与轴突切开相关的信号传导反应。我们开发了小龙虾拉伸受体分离的新显微外科手术方法，该方法保留了感觉神经元与腹神经索神经节的连接。拉伸感受器可以保留在腹部甲壳上，也可以完全隔离。在这两种情况下，它都可以是完整的，也可以是被切除的。通过从近端和远端轴突点发射记录来确认轴突的完整性。普通的，使用Hoechst 33342和碘化丙啶进行双荧光染色可观察到坏死和凋亡细胞。孤立的机械感受器神经元在8-10个小时或更长时间内保持规律的放电。与无轴突神经元的神经胶质细胞相比，无轴突神经元的神经胶质细胞的坏死和凋亡水平更低。与现有方法不同，在现有方法中将感觉神经元进行轴突切除，本方法保留了感觉神经元和神经节之间的联系，并有可能避免在神经元和附属神经胶质中进行轴索切开术的后果。本神经胶质制剂可作为简单但可提供信息的模型对象，用于研究轴突切开术引起的周围神经元变性和存活，神经胶质在神经元损伤中的作用，神经胶质相互作用的信号传导机制，