Comparison of the electrophysiological and immunohistochemical properties of acutely dissociated and 1-day cultured rat trigeminal ganglion neurons
Comparison of the electrophysiological and immunohistochemical properties of acutely dissociated and 1-day cultured rat trigeminal ganglion neurons
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急性解离和培养1天的大鼠三叉神经节神经元电生理和免疫组织化学特性的比较
DOI：
10.1016/j.neulet.2012.06.069
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发表时间：
2012
期刊：
影响因子：
2.5
通讯作者：
Inenaga K
中科院分区：
文献类型：
--
作者：
Ono K;Xu S;Hitomi S;Inenaga K
为了阐明感觉神经元的细胞特性是否在短暂培养后发生变化，我们比较了大鼠三叉神经节神经元的电生理和免疫组织化学特性。我们比较了这些神经元急性分离后，1天的无血清和无神经营养因子的条件下培养。在全细胞膜片钳记录中，培养1天的神经元需要较低的电流阈值来诱导小型和中型神经元的动作电位。此外，输入电阻较高的中型神经元培养1天后，急性分离的中型神经元相比。免疫荧光研究表明，激活转录因子3（ATF3）的易位到细胞核和低阈值Na+通道（Nav1.3）的表达上调后1天的文化。然而，在急性分离的神经元中，ATF3易位发生在低水平，Nav1.3不表达。这些电生理和免疫组化的变化后，1天的文化是非常相似的报告后发生的神经损伤的变化。我们的研究表明，损伤样特征似乎表现在1天培养的感觉神经元，这不会发生在急性分离的神经元。总的来说，我们的结果是相关的，并将有助于解释研究结果时，检查分离的感觉神经元在疼痛研究。
To clarify whether changes to the cellular properties of sensory neurons occur after a brief culture, we compared the electrophysiological and immunohistochemical properties of rat trigeminal ganglion neurons. We compared these neurons after acute dissociation and after a 1-day culture under serum-free and neurotrophin-free conditions. In whole-cell patch-clamp recordings, the 1-day cultured neurons required a lower current threshold to induce an action potential in both small- and medium-sized neurons. Furthermore, the input resistance was higher in the medium-sized neurons after a 1-day culture compared to the acutely dissociated medium-sized neurons. Immunofluorescent studies demonstrated that both the translocation of the activating transcription factor 3 (ATF3) into the nucleus and the expression of a low threshold Na+channel (Nav1.3) were upregulated after 1-day of culture. However, in the acutely dissociated neurons, ATF3 translocation occurred at low levels, and Nav1.3 was not expressed. These electrophysiological and immunohistochemical changes after 1-day of culture were very similar to the reported changes that occur after nerve injury. Our study demonstrated that injury-like characteristics appear to be manifested in the 1-day cultured sensory neurons, which do not occur in acutely dissociated neurons. Overall, our results are relevant and will help when interpreting the results of studies examining dissociated sensory neurons in pain research.