SSBD:database

Project ID

268-Oishi-VesselClassification

Project SID

268

Title

Confocal immunohistochemistry images and BDML file for quantitative information of SMA/laminin α2 mouse vessel-type classification for machine learning.

Description

Nerve/glial antigen 2 (NG2) is a protein marker of NG2 glia and mural cells, and NG2 promoter activity is utilized to target these cells. However, the NG2 promoter cannot target NG2 glia and mural cells separately. This has been an obstacle for NG2 gliaspecific manipulation. Here, we developed transgenic mice in which either cell type can be targeted using the NG2 promoter. We selected a tetracycline-controllable gene induction system for cell type-specific transgene expression, and generated NG2-tetracycline transactivator (tTA) transgenic lines. We crossed tTA lines with the tetO-ChR2 (channelrhodopsin-2)-EYFP line to characterize tTA-dependent transgene induction. We isolated two unique NG2-tTA mouse lines: one that induced ChR2-EYFP only in mural cells, likely due to the chromosomal position effect of NG2-tTA insertion, and the other that induced it in both cell types. We then applied a Cre-mediated set-subtraction strategy to the latter case and eliminated ChR2-EYFP from mural cells, resulting in NG2 glia-specific transgene induction. We further demonstrated that tTA-dependent ChR2 expression could manipulate cell function. Optogenetic mural cell activation decreased cerebral blood flow, as previously reported, indicating that tTA-mediated ChR2 expression was sufficient to impact cellular function. ChR2-mediated depolarization was observed in NG2 glia in acute hippocampal slices. In addition, ChR2-mediated depolarization of NG2 glia inhibited their proliferation but promoted their differentiation in juvenile mice. Since the tTA– tetO combination is expandable, the mural cell–specific NG2-tTA line and the NG2 glia-specific NG2-tTA line will permit us to conduct observational and manipulation studies to examine in vivo function of these cells separately.

Submission Date

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Opened Date

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Release Date

2023-09-01

Update Date

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Kind

License

Project URL

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Project DOI

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Metadata Version

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Template Version

3.0.1

Funding Information

This work was supported by Research Grants for Life Science and Medicine by Keio University Medical Science Fund to Mitsuhiro Oishi, Keio University Grant-in-Aid for Encouragement of Young Medical Scientists to Mitsuhiro Oishi, a Grant-in-Aid for Transformative Research Areas (A) “Glia decoding” (20H05896) from Japan Society for the Promotion of Science (JSPS) to Kenji F. Tanaka, JSPS Core-toCore Program, Advanced Research Networks to Kenji F. Tanaka and Christian Steinhäuser, and Japan Science and Technology Agency (JST) CREST program (JPMJCR1921) to Itaru Imayoshi.

Organism

Strain

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Cell

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Cell line

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MeSH

Anatomical Entity

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UBERON

Biological Process

Cellular Component

Molecular Function

Medical Subject Headings

Method involved in biological imaging