scRNA-seq of electrophysiologically characterized mechanosensetive DRG neurons reveals maturational differences in gene expression. scRNA-seq of electrophysiologically characterized mechanosensetive DRG neurons reveals maturational differences in gene expression

Cultured dorsal root ganglia neurons of 4 and 12 weeks old C57Bl/6 mice were first characterized as mechanosensitive (showed rapidly adapting mechanically actived currents in whole-cell votage clamp) and as mechanoreceptors (evoked action potential waveform in whole-cell current clamp). Next, cells were subjected to RNA-seq and sequences were analyzed bioinformatically. We found maturatinal changes comprising age-dependent differential gene expression in these somatosensory mechanoreceptors. The found differences call for tight age-matching of young mice in research and underline found electrophysiological and behavioural maturational changes in somatosensation, see Michel, Niklas and Narayanan, Pratibha and Shomroni, Orr and Schmidt, Manuela, Cutaneous Touch and Piezo2-Mediated Mechanotransduction Decline Throughout Mouse Maturation Until Adulthood. CELL-REPORTS-D-20-00425. Available at SSRN: https://ssrn.com/abstract=3536401 or http://dx.doi.org/10.2139/ssrn.3536401 Overall design: 18 single DRG neuron RNA profiles of 4 and 12 weeks old male WT C57Bl/6J mice, grouped by their electrophysiological profile

本研究首先对4周龄与12周龄C57BL/6小鼠的培养背根神经节（dorsal root ganglia, DRG）神经元进行表型鉴定：通过全细胞膜片钳（whole-cell voltage clamp）记录到其具备机械敏感性，可呈现快速适应性机械激活电流；同时可被鉴定为机械感受器，即在全细胞电流钳（whole-cell current clamp）模式下可诱发出动作电位波形。随后，对上述神经元开展RNA测序（RNA-seq），并通过生物信息学方法对测序序列进行分析。本研究在这些躯体感觉机械感受器中发现了随年龄变化的差异基因表达所构成的成熟性改变。上述差异提示，相关研究需严格匹配幼年小鼠的年龄，同时也进一步佐证了躯体感觉系统在电生理与行为学层面的成熟性改变。相关研究详见：Michel, Niklas、Narayanan, Pratibha、Shomroni, Orr、Schmidt, Manuela所撰论文《Cutaneous Touch and Piezo2-Mediated Mechanotransduction Decline Throughout Mouse Maturation Until Adulthood》，文献编号为CELL-REPORTS-D-20-00425，可于SSRN平台获取：https://ssrn.com/abstract=3536401 或 http://dx.doi.org/10.2139/ssrn.3536401。实验整体设计：选取4周龄及12周龄的雄性野生型（wild type, WT）C57BL/6J小鼠，共获得18份单个背根神经节神经元的RNA测序谱，并根据其电生理特征完成分组。