Division of Physiology, Kyushu Dental College, 2-6-1 Manazuru, Kokurakitaku, Kitakyushu 803-8580, Japan.
Neurosci Lett. 2012 Aug 15;523(2):162-6. doi: 10.1016/j.neulet.2012.06.069. Epub 2012 Jul 4.
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 (Na(v)1.3) were upregulated after 1-day of culture. However, in the acutely dissociated neurons, ATF3 translocation occurred at low levels, and Na(v)1.3 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.
为了阐明感觉神经元的细胞特性在短暂培养后是否会发生变化,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。我们比较了急性分离后和在无血清和无神经生长因子条件下培养 1 天后的这些神经元。在全细胞膜片钳记录中,培养 1 天的神经元在小和中型神经元中引起动作电位所需的电流阈值较低。此外,与急性分离的中型神经元相比,培养 1 天后中型神经元的输入电阻更高。免疫荧光研究表明,激活转录因子 3(ATF3)向核内易位和低阈值钠离子通道(Na(v)1.3)的表达在培养 1 天后均上调。然而,在急性分离的神经元中,ATF3 易位水平较低,Na(v)1.3 不表达。培养 1 天后的这些电生理和免疫组织化学变化与神经损伤后发生的变化非常相似。我们的研究表明,类似损伤的特征似乎在培养 1 天的感觉神经元中表现出来,而在急性分离的神经元中则不会出现。总的来说,我们的研究结果具有相关性,有助于解释在疼痛研究中研究分离的感觉神经元的研究结果。