López-Hernández Gretchen Y, Ananth Mala, Jiang Li, Ballinger Elizabeth C, Talmage David A, Role Lorna W
Department of Neurobiology and Behavior, Stony Brook University, Stony Brook, NY, USA.
Center for Nervous System Disorders, Stony Brook University, Stony Brook, NY, USA.
J Neurochem. 2017 Aug;142 Suppl 2(Suppl 2):103-110. doi: 10.1111/jnc.14073.
Recent developments in the generation of neuronal population-specific, genetically modified mouse lines have allowed precise identification and selective stimulation of cholinergic neurons in vivo. Although considerably less laborious than studies conducted with post hoc identification of cholinergic neurons by immunostaining, it is not known whether the genetically based labeling procedures that permit in vivo identification are electrophysiologically benign. In this study, we use mice carrying a bacterial artificial chromosome transgene that drives expression of a tau-green fluorescent fusion protein specifically in cholinergic neurons. This allowed us to visualize basal forebrain cholinergic neurons in acute slice preparations. Using whole cell, patch clamp electrophysiological recording in acute brain slices, here we present original data about the basic electrical properties of these genetically tagged cholinergic neurons including firing rate, resting membrane potential, rheobase, and various characteristics of their action potentials and after-hyperpolarization potentials. The basic electrical properties are compared (i) with non-cholinergic neurons in the same brain regions; (ii) in cholinergic neurons between immature animals and young adults; and (iii) with cholinergic neurons that are expressing light-sensitive channels. Our conclusions based on these data are (i) cholinergic neurons are less excitable then their non-cholinergic neighbors, (ii) the basic properties of cholinergic neurons do not significantly change between adolescence and young adulthood and (iii) these properties are not significantly affected by chronic expression of the excitatory opsin, oChIEF. This is an article for the special issue XVth International Symposium on Cholinergic Mechanisms.
神经元群体特异性转基因小鼠品系生成技术的最新进展,使得在体内精确识别和选择性刺激胆碱能神经元成为可能。尽管这比通过免疫染色对胆碱能神经元进行事后识别的研究要省力得多,但尚不清楚允许在体内识别的基于基因的标记程序在电生理方面是否无害。在本研究中,我们使用携带细菌人工染色体转基因的小鼠,该转基因驱动tau-绿色荧光融合蛋白在胆碱能神经元中特异性表达。这使我们能够在急性脑片制备中可视化基底前脑胆碱能神经元。在此,我们通过在急性脑片中进行全细胞、膜片钳电生理记录,展示了关于这些基因标记的胆碱能神经元基本电生理特性的原始数据,包括放电频率、静息膜电位、阈强度以及它们动作电位和超极化后电位的各种特征。将这些基本电生理特性进行了如下比较:(i)与同一脑区的非胆碱能神经元进行比较;(ii)在未成熟动物和年轻成年动物的胆碱能神经元之间进行比较;(iii)与表达光敏感通道的胆碱能神经元进行比较。基于这些数据我们得出的结论是:(i)胆碱能神经元的兴奋性低于其相邻的非胆碱能神经元;(ii)胆碱能神经元的基本特性在青春期和年轻成年期之间没有显著变化;(iii)这些特性不受兴奋性视蛋白oChIEF慢性表达的显著影响。这是一篇发表于第十五届国际胆碱能机制研讨会特刊的文章。