p2a cre  (Addgene inc)

Journal: bioRxiv

Article Title: Sphingosine-1-Phosphate Receptor 1 regulates competition dependent astrocyte morphogenesis and tiling in murine cortex

doi: 10.64898/2026.03.28.714989

Figure Lengend Snippet: (A) Schematics showing S1PR1 deletion in sparse astrocytes by delivering AAV-GfaABC1D-YFP-P2A-Cre or AAV-GfaABC1D-YFP control in S1PR1 fl/fl mouse pups. (B) Sparsely labeled astrocytes were imaged from L2-3 somatosensory cortex from P30 pups. ( C ) Representative confocal images and subsequent IMARIS 3D surface/filament renderings of YFP astrocytes from S1PR1 ΔAST or littermate controls at P30. (D) Quantification of astrocyte volume (left) and soma volume (right) from Surface renderings generated by IMARIS. ( E ) Quantification of astrocyte territory volume (left) filament length (middle) and filament area (right) from filament traces generated by IMARIS. (F) Plot depicting the proportion of astrocyte filaments grouped by various mean diameter and represented as % of total filaments revealed no differences in mean dendrites in S1PR1 ΔAST astrocytes. ( G ) Sholl analyses of IMARIS rendered filament traces of YFP labeled sparse astrocytes from S1PR1 ΔAST and littermate controls. Data represents the mean ± SEM of 23 and 24 astrocytes from n=4 mice per group. * = p <0.05, ** = p <0.01, ns=non-significant, Unpaired Welch’s T-test.

Article Snippet: All AAVs used in this study were produced in the Singh lab unless otherwise mentioned, in accordance with the US National Institutes of Health Guidelines for Research Involving Recombinant DNA Molecules and the Virginia Commonwealth University Institutional Biosafety Committee. pAAV-GfaBC1D-YFP-P2A-Cre plasmid was created by subcloning P2A-Cre from pAAV-hSynapsin-mCherry-P2A-Cre-WPRE plasmid (a gift from Hui Yang (Addgene plasmid # 107312); into the pAAV-GfaBC1D-YFP plasmid.

Techniques: Control, Labeling, Generated

Journal: bioRxiv

Article Title: Sphingosine-1-Phosphate Receptor 1 regulates competition dependent astrocyte morphogenesis and tiling in murine cortex

doi: 10.64898/2026.03.28.714989

Figure Lengend Snippet: (A) Schematics showing S1PR1 deletion in sparse astrocytes by delivering AAV-GfaABC1D-YFP-P2A-Cre or AAV-GfaABC1D-YFP control in S1PR1 fl/fl mouse pups. Sparsely labeled astrocytes were imaged from L4-5 somatosensory cortex from P30 pups. ( B ) Representative confocal images and subsequent IMARIS 3D surface/filament renderings of YFP astrocytes from S1PR1 ΔAST or littermate controls at P30. (C) Quantification of astrocyte volume (left) and soma volume (right) from Surface renderings generated by IMARIS. ( E ) Quantification of astrocyte territory volume (left) process/filament length (middle) and area (left) from filament traces generated by IMARIS. (F) Plot depicting the proportion of astrocyte filaments grouped by mean diameter and represented as % of total filaments revealed no differences in mean dendrites in S1PR1 ΔAST astrocytes. ( G ) Sholl analyses of IMARIS rendered filament traces of GFP labeled sparse astrocytes from S1PR1 ΔAST and littermate controls. Data represents the mean ± SEM of 22 and 17 astrocytes from n=4 mice per group. * = p <0.05, ** = p <0.01, *** = p <0.001 ns=non-significant, Unpaired Welch’s T-test.

Article Snippet: All AAVs used in this study were produced in the Singh lab unless otherwise mentioned, in accordance with the US National Institutes of Health Guidelines for Research Involving Recombinant DNA Molecules and the Virginia Commonwealth University Institutional Biosafety Committee. pAAV-GfaBC1D-YFP-P2A-Cre plasmid was created by subcloning P2A-Cre from pAAV-hSynapsin-mCherry-P2A-Cre-WPRE plasmid (a gift from Hui Yang (Addgene plasmid # 107312); into the pAAV-GfaBC1D-YFP plasmid.

Techniques: Control, Labeling, Generated

Journal: bioRxiv

Article Title: Sphingosine-1-Phosphate Receptor 1 regulates competition dependent astrocyte morphogenesis and tiling in murine cortex

doi: 10.64898/2026.03.28.714989

Figure Lengend Snippet: B) Schematics showing S1PR1 deletion in neighboring astrocytes in two-color schemes by delivering AAV-GfaABC1D-YFP-P2A-Cre plus AAV-GfaABC1D-tdTomato-P2A-Cre or AAV-GfaABC1D-YFP plus AAV-GfaABC1D-tdTomato controls in S1PR1 fl/fl mouse pups. ( C ) Example 20x confocal image to show the labelling by two color viruses. (D) Representative confocal images and subsequent IMARIS 3D surface/filament renderings to create territories of neighboring astrocytes from KO:KO or littermate WT:WT controls at P30. (E) Quantification of astrocyte territory overlap volume represented as % of total volume from two neighboring astrocytes. Data represents the mean ± SEM of 23 and 18 pairs of neighboring astrocytes from n=3 and 4 mice per group. **** = p <0.0001, Unpaired Welch’s T-test.

Article Snippet: All AAVs used in this study were produced in the Singh lab unless otherwise mentioned, in accordance with the US National Institutes of Health Guidelines for Research Involving Recombinant DNA Molecules and the Virginia Commonwealth University Institutional Biosafety Committee. pAAV-GfaBC1D-YFP-P2A-Cre plasmid was created by subcloning P2A-Cre from pAAV-hSynapsin-mCherry-P2A-Cre-WPRE plasmid (a gift from Hui Yang (Addgene plasmid # 107312); into the pAAV-GfaBC1D-YFP plasmid.

Techniques: