Detail of NG2BChR2_2516_GFPSMAa2_20X_cortex



Project
Title
Confocal immunoshistochemistry images of SMA/laminin (alpha)2 mouse vessel-type classification for machine learning.
Description
Confocal immunoshistochemistry images of SMA/laminin (alpha)2 mouse vessel-type classification for machine learning.
Release, Updated
2023-09-01
License
CC BY
Kind
Image data
File Formats
.oir
Data size
276.5 MB

Organism
Mus musculus ( NCBI:txid10090 )
Strain(s)
STOCK
Cell Line
-

Datatype
-
Molecular Function (MF)
laminin binding ( GO:0043236 )
Biological Process (BP)
arteriole smooth muscle contraction ( GO:0014830 )
Cellular Component (CC)
laminin-2 complex ( GO:0005607 )
Biological Imaging Method
confocal microscopy ( Fbbi:00000251 )
X scale
0.6214806 micrometer/pixel
Y scale
0.6214806 micrometer/pixel
Z scale
0.960 micrometer/slice
T scale
-

Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
-
Objective model
-
Filter set
-

Summary of Methods
See details in Oishi M, et. al. (2023) Glia. Feb;71(2):317-333.
Related paper(s)

Mitsuhiro Oishi, Stefan Passlick, Yoshihiko Yamazaki, Miyuki Unekawa, Ruka Adachi, Mayumi Yamada, Itaru Imayoshi, Yoshifumi Abe, Christian Steinhauser, Kenji F Tanaka (2022) Separate optogenetic manipulation of Nerve/glial antigen 2 (NG2) glia and mural cells using the NG2 promoter., Glia

Published in 2022 Sep 27 (Electronic publication in Sept. 27, 2022, midnight )

(Abstract) 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 glia-specific 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.

Contact
Kenji F Tanaka , Keio University School of Medicine , Division of Brain Sciences
Contributors
Mitsuhiro OishiMitsuhiro Oishi

OMERO Dataset
OMERO Project
Source