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

doi:10.16098/j.issn.0529-1356.2025.05.001

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Acta Anatomica Sinica ›› 2025, Vol. 56 ›› Issue (5) : 507-514. DOI: 10.16098/j.issn.0529-1356.2025.05.001

Neurbiology

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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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.

Key words

Oligodendrocyte precursor cell / Chicken opsin 5 / G protein-coupled receptor / Calcium imaging / Optogenetics / Mouse

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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

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