Department of Neuroscience II, Research Institute of Environmental Medicine, Nagoya University, Nagoya, 464-8601, Japan.
Department of Neural Regulation, Nagoya University Graduate School of Medicine, Nagoya, 466-8550, Japan.
Sci Rep. 2023 Mar 9;13(1):3966. doi: 10.1038/s41598-023-30907-2.
Optogenetics is a powerful tool for manipulating neuronal activity by light illumination with high temporal and spatial resolution. Anion-channelrhodopsins (ACRs) are light-gated anion channels that allow researchers to efficiently inhibit neuronal activity. A blue light-sensitive ACR2 has recently been used in several in vivo studies; however, the reporter mouse strain expressing ACR2 has not yet been reported. Here, we generated a new reporter mouse strain, LSL-ACR2, in which ACR2 is expressed under the control of Cre recombinase. We crossed this strain with a noradrenergic neuron-specific driver mouse (NAT-Cre) to generate NAT-ACR2 mice. We confirmed Cre-dependent expression and function of ACR2 in the targeted neurons by immunohistochemistry and electrophysiological recordings in vitro, and confirmed physiological function using an in vivo behavioral experiment. Our results show that the LSL-ACR2 mouse strain can be applied for optogenetic inhibition of targeted neurons, particularly for long-lasting continuous inhibition, upon crossing with Cre-driver mouse strains. The LSL-ACR2 strain can be used to prepare transgenic mice with homogenous expression of ACR2 in targeted neurons with a high penetration ratio, good reproducibility, and no tissue invasion.
光遗传学是一种通过光照明以高时间和空间分辨率来操纵神经元活动的强大工具。阴离子通道视蛋白（ACRs）是光门控阴离子通道，使研究人员能够有效地抑制神经元活动。最近，一种对蓝光敏感的 ACR2 已在几项体内研究中使用；然而，表达 ACR2 的报告小鼠品系尚未报道。在这里，我们生成了一种新的报告小鼠品系，LSL-ACR2，其中 ACR2 在 Cre 重组酶的控制下表达。我们将该品系与去甲肾上腺素能神经元特异性驱动小鼠（NAT-Cre）杂交，生成 NAT-ACR2 小鼠。我们通过免疫组织化学和体外电生理记录证实了 ACR2 在靶向神经元中的 Cre 依赖性表达和功能，并通过体内行为实验证实了其生理功能。我们的结果表明，LSL-ACR2 小鼠品系可与 Cre 驱动小鼠品系杂交，用于靶向神经元的光遗传学抑制，特别是用于持久的连续抑制。LSL-ACR2 品系可用于制备在靶向神经元中具有高穿透比、良好重现性且无组织侵袭的同质 ACR2 表达的转基因小鼠。
