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首页> 外文期刊>Romanian Journal of Biophysics >CLASSIFICATION OF DORSAL ROOT GANGLION NEURONS FROM NEWBORN RAT IN ORGANOTYPIC CULTURES
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CLASSIFICATION OF DORSAL ROOT GANGLION NEURONS FROM NEWBORN RAT IN ORGANOTYPIC CULTURES

机译：新生大鼠背根神经节神经元在有机培养中的分类

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This work proposes an in vitro model to study different types of ionic channels in primary sensory neurons involved in nociception. Using dorsal root ganglia (DRG) slices from newborn rats maintained in culture on three types of substrates, we have tested neuronal viability by two distinct methods: a stereological (physical dissector) and an electrophysiological method (patch-clamp technique, whole cell configuration). After applying viability tests, we have chosen to maintain DRG slices in culture on collagen-coated Petri dishes. This substrate preserved a high (80–90%) neuronal viability after 10 days in culture, and was easy to prepare. For subclassification of DRG neurons using current signature protocols, slices were maintained in culture for either 2 or 5 days. In addition, to differentiate neurons belonging to nociceptive classes, we analyzed some characteristics of action potentials (duration at 0 mV, at return to baseline, at –60 mV, and at 80% recovery from afterhyperpolarization (AHP 80%)). Following the criteria of Petruska et al. [32], we found after 2 days culture only types 1, 3, 4, 5, 7, 8 and 9 neurons. Cultures at 5 days lacked the types 2, 5, 6, 7 and 8 of cells. The magnitude of hyperpolarization-activated currents (HACs) obtained by us in DRG neurons from newborn rats was small (–94 ± 1.1 pA), confirming that primary sensory neurons at this developmental age feature low levels of HAC expression.
机译：这项工作提出了一种体外模型，用于研究参与伤害感受的初级感觉神经元中不同类型的离子通道。我们使用新生大鼠的背根神经节（DRG）切片在三种类型的基质上培养，我们通过两种不同的方法测试了神经元的生存力：一种立体方法（物理解剖器）和一种电生理方法（膜片钳技术，全细胞配置） 。应用可行性测试后，我们选择将DRG切片在胶原蛋白覆盖的培养皿上进行培养。培养10天后，该底物保留了较高的（80-90％）神经元活力，并且易于制备。为了使用当前的签名方案对DRG神经元进行亚分类，将切片在培养物中保持2或5天。此外，为了区分属于伤害感受类的神经元，我们分析了动作电位的一些特征（持续时间为0 mV，恢复到基线时，–60 mV时以及从超极化后恢复80％（AHP 80％））。遵循Petruska等人的标准。 [32]，我们发现在培养2天后仅培养了1、3、4、5、7、8和9型神经元。 5天的培养物缺乏细胞的2、5、6、7和8型。我们在新生大鼠的DRG神经元中获得的超极化激活电流（HAC）的大小很小（–94±1.1 pA），这证实了在这个发育年龄的初级感觉神经元的HAC表达水平较低。

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