光特异性激活少突胶质前体细胞模型小鼠的构建与验证

doi:10.16098/j.issn.0529-1356.2025.05.001

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解剖学报 ›› 2025, Vol. 56 ›› Issue (5) : 507-514. DOI: 10.16098/j.issn.0529-1356.2025.05.001

神经生物学

光特异性激活少突胶质前体细胞模型小鼠的构建与验证

王舒悦¹ 申杨贝纳² 黄南昕¹ 李思维¹于彬² 王玉鑫^3* 肖岚^2*

作者信息 +

Construction and validation of a mouse model for optically activation of oligodendrocyte precursor cells

WANG Shu-yue¹ SHENYANG Bei-na² HUANG Nan-xin¹ LI Si-wei¹ YU Bin² WANG Yu-xin^3* XIAO Lan ^2*

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文章历史 +

摘要

目的构建一种可特异性光激活少突胶质前体细胞（OPCs）的转基因小鼠并进行验证。方法利用CRISPR/Cas9基因编辑技术构建鸡视蛋白5(cOpn5) Rosa26-LSL-cOpn5品系小鼠，并将其与NG2-CreERT品系小鼠进行交配获得NG2-CreERT;cOpn5转基因小鼠，该模型小鼠经他莫昔芬诱导后可实现光敏蛋白cOpn5在OPCs中的特异性表达。利用免疫荧光染色对该转基因小鼠转入效率和特异性进行验证。进一步利用急性脑片钙成像验证470nm蓝光激活OPCs的功能效应。结果在NG2-CreERT；cOpn5转基因小鼠中，OPCs可特异性表达光敏蛋白cOpn5。急性脑片钙成像结果显示，蓝光刺激可引起NG2-CreERT；cOpn5光遗传小鼠细胞钙活动增强，表明OPCs可被光激活。结论成功构建了一种可特异性光激活OPCs的转基因模型小鼠，为后续在体研究OPCs在中枢神经系统中的功能和调控机制提供了新的工具。

Abstract

Objective To develop and validate a transgenic mouse model enabling specific and inducible optogenetic activation of oligodendrocyte precursor cells (OPCs). Methods A conditional allele for the photosensitive opsin chicken opsin 5(cOpn5) (Rosa26-LSL-cOpn5) was generated using CRISPR/Cas9 technology. These mice were subsequently crossed with NG2-CreERT transgenic mice to produce NG2-CreERT;cOpn5 animals. In this model, tamoxifen administration induces Cre-mediated recombination, leading to specific expression of cOpn5 in NG2-positive OPCs. The specificity and efficiency of cOpn5 expression in OPCs were confirmed by immunofluorescent staining. Functional validation of light-induced OPC activation was performed by using calcium imaging in acute brain slices after stimulation with 470nm blue light. Results Immunofluorescence analysis confirmed robust and specific expression of cOpn5 within NG2-positive OPCs in the brains of tamoxifentreated NG2-CreERT;cOpn5 mice. Crucially, calcium imaging of acute brain slices from these mice demonstrated a significant increase in intracellular calcium levels in cOpn5-expressing OPCs upon stimulation with 470nm blue light, indicating successful optogenetic activation. Conclusion We have successfully generated and validated a novel transgenic mouse model (NG2-CreERT;cOpn5) that permits specific and inducible optogenetic activation of OPCs. This model provides a novel tool for subsequent in vivo studies of the role and regulating mechanisms of OPCs in the central nervous system.

导出引用

王舒悦申杨贝纳黄南昕李思维于彬王玉鑫肖岚. 光特异性激活少突胶质前体细胞模型小鼠的构建与验证[J]. 解剖学报. 2025, 56(5): 507-514 https://doi.org/10.16098/j.issn.0529-1356.2025.05.001

WANG Shu-yue SHENYANG Bei-na HUANG Nan-xin LI Si-wei YU Bin WANG Yu-xin XIAO Lan. Construction and validation of a mouse model for optically activation of oligodendrocyte precursor cells[J]. Acta Anatomica Sinica. 2025, 56(5): 507-514 https://doi.org/10.16098/j.issn.0529-1356.2025.05.001

中图分类号： R322.8 Q95-33

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