Laboratory of Glia Biology


Identification of region-specific astrocyte subtypes at single cell resolution
Mykhailo Y. Batiuk, Araks Martirosyan, Jérôme Wahis, Filip de Vin, Catherine Marneffe, Carola Kusserow, Jordan Koeppen, João Filipe Viana, João Filipe Oliveira, Thierry Voet, Chris P. Ponting, T. Grant Belgard, Matthew G. Holt, Nature Communications volume 11, Article number: 1220 (2020)
BioRxiv preprint May 2018; doi: http://dx.doi.org/10.1101/317503
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Differential patterning of cells in adult mouse brain mapped with ISH identified by Gfap+Agt+Slc1a3+ marker combination in adult mouse brain. Mapping is done based on multiplex ISH preformed on brain sections from animals of age P56-P60. Top left is an example of a brain section with the mirrored reference atlas. Black dots on the section show putative location of a subtype identified by co-localization of corresponding biomarker (scale bar, 1000μm). In the boxes below markers used to define a subtype are shown separately (on the top of the DAPI signal in blue) followed by a merged image (scale bar, 10μm). Barplots on the right show in situ single cell RNA marker counts per astrocytes (Slc1a3+ cells) followed by the dis-tribution of the subtype among brain regions and the normaized distribution of the subtype by the number of astrocytes (Glast encoding Slc1a3+ cells) per brain region. In the barplots astrocytes belonging to the subtype of interest are highlighted with the corresponding color, counts per marker are sorted in the descending order starting from the right. Error bars represent SEM averaged across all stained sections.

Differential patterning of cells in adult mouse brain mapped with ISH identified by Unc13c+Agt-Slc1a3+ marker combination in adult mouse brain. Mapping is done based on multiplex ISH preformed on brain sections from animals of age P56-P60. Top left is an example of a brain section with the mirrored reference atlas. Black dots on the section show putative location of a subtype identified by co-localization of corresponding biomarker (scale bar, 1000μm). In the boxes below markers used to define a subtype are shown separately (on the top of the DAPI signal in blue) followed by a merged image (scale bar, 10μm). Barplots on the right show in situ single cell RNA marker counts per astrocytes (Slc1a3+ cells) followed by the dis-tribution of the subtype among brain regions and the normaized distribution of the subtype by the number of astrocytes (Glast encoding Slc1a3+ cells) per brain region. In the barplots astrocytes belonging to the subtype of interest are highlighted with the corresponding color, counts per marker are sorted in the descending order starting from the right. Error bars represent SEM averaged across all stained sections.

Differential patterning of cells in adult mouse brain mapped with ISH identified by Gfap-Agt+Slc1a3+ marker combination in adult mouse brain. Mapping is done based on multiplex ISH preformed on brain sections from animals of age P56-P60. Top left is an example of a brain section with the mirrored reference atlas. Black dots on the section show putative location of a subtype identified by co-localization of corresponding biomarker (scale bar, 1000μm). In the boxes below markers used to define a subtype are shown separately (on the top of the DAPI signal in blue) followed by a merged image (scale bar, 10μm). Barplots on the right show in situ single cell RNA marker counts per astrocytes (Slc1a3+ cells) followed by the dis-tribution of the subtype among brain regions and the normaized distribution of the subtype by the number of astrocytes (Glast encoding Slc1a3+ cells) per brain region. In the barplots astrocytes belonging to the subtype of interest are highlighted with the corresponding color, counts per marker are sorted in the descending order starting from the right. Error bars represent SEM averaged across all stained sections.

Differential patterning of cells in adult mouse brain mapped with ISH identified by Unc13c-Agt+Slc1a3+ marker combination in adult mouse brain. Mapping is done based on multiplex ISH preformed on brain sections from animals of age P56-P60. Top left is an example of a brain section with the mirrored reference atlas. Black dots on the section show putative location of a subtype identified by co-localization of corresponding biomarker (scale bar, 1000μm). In the boxes below markers used to define a subtype are shown separately (on the top of the DAPI signal in blue) followed by a merged image (scale bar, 10μm). Barplots on the right show in situ single cell RNA marker counts per astrocytes (Slc1a3+ cells) followed by the dis-tribution of the subtype among brain regions and the normaized distribution of the subtype by the number of astrocytes (Glast encoding Slc1a3+ cells) per brain region. In the barplots astrocytes belonging to the subtype of interest are highlighted with the corresponding color, counts per marker are sorted in the descending order starting from the right. Error bars represent SEM averaged across all stained sections.

Differential patterning of cells in adult mouse brain mapped with ISH identified by Frzb+Ascl1+Slc1a3+ marker combination in adult mouse brain. Mapping is done based on multiplex ISH preformed on brain sections from animals of age P56-P60. Top left is an example of a brain section with the mirrored reference atlas. Black dots on the section show putative location of a subtype identified by co-localization of corresponding biomarker (scale bar, 1000μm). In the boxes below markers used to define a subtype are shown separately (on the top of the DAPI signal in blue) followed by a merged image (scale bar, 10μm). Barplots on the right show in situ single cell RNA marker counts per astrocytes (Slc1a3+ cells) followed by the dis-tribution of the subtype among brain regions and the normaized distribution of the subtype by the number of astrocytes (Glast encoding Slc1a3+ cells) per brain region. In the barplots astrocytes belonging to the subtype of interest are highlighted with the corresponding color, counts per marker are sorted in the descending order starting from the right. Error bars represent SEM averaged across all stained sections.

Differential patterning of cells in adult mouse brain mapped with ISH identified by Frzb+(Nes+)Slc1a3+ marker combination in adult mouse brain. Mapping is done based on multiplex ISH preformed on brain sections from animals of age P56-P60. Top left is an example of a brain section with the mirrored reference atlas. Black dots on the section show putative location of a subtype identified by co-localization of corresponding biomarker (scale bar, 1000μm). In the boxes below markers used to define a subtype are shown separately (on the top of the DAPI signal in blue) followed by a merged image (scale bar, 10μm). Barplots on the right show in situ single cell RNA marker counts per astrocytes (Slc1a3+ cells) followed by the dis-tribution of the subtype among brain regions and the normaized distribution of the subtype by the number of astrocytes (Glast encoding Slc1a3+ cells) per brain region. In the barplots astrocytes belonging to the subtype of interest are highlighted with the corresponding color, counts per marker are sorted in the descending order starting from the right. Error bars represent SEM averaged across all stained sections.

Differential patterning of cells in adult mouse brain mapped with ISH identified by Ogt-Fam107a+Slc1a3+ marker combination in adult mouse brain. Mapping is done based on multiplex ISH preformed on brain sections from animals of age P56-P60. Top left is an example of a brain section with the mirrored reference atlas. Black dots on the section show putative location of a subtype identified by co-localization of corresponding biomarker (scale bar, 1000μm). In the boxes below markers used to define a subtype are shown separately (on the top of the DAPI signal in blue) followed by a merged image (scale bar, 10μm). Barplots on the right show in situ single cell RNA marker counts per astrocytes (Slc1a3+ cells) followed by the dis-tribution of the subtype among brain regions and the normaized distribution of the subtype by the number of astrocytes (Glast encoding Slc1a3+ cells) per brain region. In the barplots astrocytes belonging to the subtype of interest are highlighted with the corresponding color, counts per marker are sorted in the descending order starting from the right. Error bars represent SEM averaged across all stained sections.

Differential patterning of cells in adult mouse brain mapped with ISH identified by Mfge8+Slc1a3+ marker combination in adult mouse brain. Mapping is done based on multiplex ISH preformed on brain sections from animals of age P56-P60. Top left is an example of a brain section with the mirrored reference atlas. Black dots on the section show putative location of a subtype identified by co-localization of corresponding biomarker (scale bar, 1000μm). In the boxes below markers used to define a subtype are shown separately (on the top of the DAPI signal in blue) followed by a merged image (scale bar, 10μm). Barplots on the right show in situ single cell RNA marker counts per astrocytes (Slc1a3+ cells) followed by the dis-tribution of the subtype among brain regions and the normaized distribution of the subtype by the number of astrocytes (Glast encoding Slc1a3+ cells) per brain region. In the barplots astrocytes belonging to the subtype of interest are highlighted with the corresponding color, counts per marker are sorted in the descending order starting from the right. Error bars represent SEM averaged across all stained sections.

Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 1 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 2 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.
Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST1) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST2) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST3) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST4) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap+Agt+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c+Agt-Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Gfap-Agt+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Unc13c-Agt+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+Ascl1+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Frzb+(Nes+)Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Ogt-Fam107a+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Black dots in the figure show the location of Mfge8+Slc1a3+ cells (AST5) in the left (top) and right (bottom) hemispheres of Animal 3 (a coronal section) with mirrored reference atlas. Scale bar, 1000μm.

Figure shows an overview of adult mouse brain stained for Gfap+Agt+Slc1a3+ to detect AST1 in the brain.

Figure shows an overview of adult mouse brain stained for Unc13c+Agt-Slc1a3+ to detect AST1 in the brain.

Figure shows an overview of adult mouse brain stained for Gfap-Agt+Slc1a3+ to detect AST1 in the brain.

Figure shows an overview of adult mouse brain stained for Unc13c-Agt+Slc1a3+ to detect AST1 in the brain.

Figure shows an overview of adult mouse brain stained for Frzb+Ascl1+Slc1a3+ to detect AST1 in the brain.

Figure shows an overview of adult mouse brain stained for Frzb+(Nes+)Slc1a3+ to detect AST1 in the brain.

Figure shows an overview of adult mouse brain stained for Ogt-Fam107a+Slc1a3+ to detect AST1 in the brain.

Figure shows an overview of adult mouse brain stained for Mfge8+Slc1a3+ to detect AST1 in the brain.

Figure shows an overview of adult mouse brain stained for Gfap+Agt+Slc1a3+ to detect AST2 in the brain.

Figure shows an overview of adult mouse brain stained for Unc13c+Agt-Slc1a3+ to detect AST2 in the brain.

Figure shows an overview of adult mouse brain stained for Gfap-Agt+Slc1a3+ to detect AST2 in the brain.

Figure shows an overview of adult mouse brain stained for Unc13c-Agt+Slc1a3+ to detect AST2 in the brain.

Figure shows an overview of adult mouse brain stained for Frzb+Ascl1+Slc1a3+ to detect AST2 in the brain.

Figure shows an overview of adult mouse brain stained for Frzb+(Nes+)Slc1a3+ to detect AST2 in the brain.

Figure shows an overview of adult mouse brain stained for Ogt-Fam107a+Slc1a3+ to detect AST2 in the brain.

Figure shows an overview of adult mouse brain stained for Mfge8+Slc1a3+ to detect AST2 in the brain.

Figure shows an overview of adult mouse brain stained for Gfap+Agt+Slc1a3+ to detect AST3 in the brain.

Figure shows an overview of adult mouse brain stained for Unc13c+Agt-Slc1a3+ to detect AST3 in the brain.

Figure shows an overview of adult mouse brain stained for Gfap-Agt+Slc1a3+ to detect AST3 in the brain.

Figure shows an overview of adult mouse brain stained for Unc13c-Agt+Slc1a3+ to detect AST3 in the brain.

Figure shows an overview of adult mouse brain stained for Frzb+Ascl1+Slc1a3+ to detect AST3 in the brain.

Figure shows an overview of adult mouse brain stained for Frzb+(Nes+)Slc1a3+ to detect AST3 in the brain.

Figure shows an overview of adult mouse brain stained for Ogt-Fam107a+Slc1a3+ to detect AST3 in the brain.

Figure shows an overview of adult mouse brain stained for Mfge8+Slc1a3+ to detect AST3 in the brain.

Figure shows an overview of adult mouse brain stained for Gfap+Agt+Slc1a3+ to detect AST4 in the brain.

Figure shows an overview of adult mouse brain stained for Unc13c+Agt-Slc1a3+ to detect AST4 in the brain.

Figure shows an overview of adult mouse brain stained for Gfap-Agt+Slc1a3+ to detect AST4 in the brain.

Figure shows an overview of adult mouse brain stained for Unc13c-Agt+Slc1a3+ to detect AST4 in the brain.

Figure shows an overview of adult mouse brain stained for Frzb+Ascl1+Slc1a3+ to detect AST4 in the brain.

Figure shows an overview of adult mouse brain stained for Frzb+(Nes+)Slc1a3+ to detect AST4 in the brain.

Figure shows an overview of adult mouse brain stained for Ogt-Fam107a+Slc1a3+ to detect AST4 in the brain.

Figure shows an overview of adult mouse brain stained for Mfge8+Slc1a3+ to detect AST4 in the brain.

Figure shows an overview of adult mouse brain stained for Gfap+Agt+Slc1a3+ to detect AST5 in the brain.

Figure shows an overview of adult mouse brain stained for Unc13c+Agt-Slc1a3+ to detect AST5 in the brain.

Figure shows an overview of adult mouse brain stained for Gfap-Agt+Slc1a3+ to detect AST5 in the brain.

Figure shows an overview of adult mouse brain stained for Unc13c-Agt+Slc1a3+ to detect AST5 in the brain.

Figure shows an overview of adult mouse brain stained for Frzb+Ascl1+Slc1a3+ to detect AST5 in the brain.

Figure shows an overview of adult mouse brain stained for Frzb+(Nes+)Slc1a3+ to detect AST5 in the brain.

Figure shows an overview of adult mouse brain stained for Ogt-Fam107a+Slc1a3+ to detect AST5 in the brain.

Figure shows an overview of adult mouse brain stained for Mfge8+Slc1a3+ to detect AST5 in the brain.