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Saturday, December 10, 2016
|The hippocampus of a mouse with the artificial Cre/ERT2 gene, showing astrocytes glowing green. Neurosciencenews image is credited to Baljit Khakh lab/David Geffen School of Medicine at UCLA.|
Source: David Olmos – UCLA
New Transgenic Mouse Lines for Selectively Targeting Astrocytes and Studying Calcium Signals in Astrocyte Processes In Situ and In Vivo
•Cre/ERT2 mice were made to achieve astrocyte-specific genetic manipulations in vivo
•Knockin Lck-GCaMP6f mice were made to study astrocyte calcium signals in vivo
•Mice were used to determine the adult cortical astrocyte transcriptome
•New, well-characterized, and much needed in vivo genetic resources are provided
Astrocytes exist throughout the nervous system and are proposed to affect neural circuits and behavior. However, studying astrocytes has proven difficult because of the lack of tools permitting astrocyte-selective genetic manipulations. Here, we report the generation of Aldh1l1-Cre/ERT2 transgenic mice to selectively target astrocytes in vivo. We characterized Aldh1l1-Cre/ERT2 mice using imaging, immunohistochemistry, AAV-FLEX-GFP microinjections, and crosses to RiboTag, Ai95, and new Cre-dependent membrane-tethered Lck-GCaMP6f knockin mice that we also generated. Two to three weeks after tamoxifen induction, Aldh1l1-Cre/ERT2 selectively targeted essentially all adult (P80) brain astrocytes with no detectable neuronal contamination, resulting in expression of cytosolic and Lck-GCaMP6f, and permitting subcellular astrocyte calcium imaging during startle responses in vivo. Crosses with RiboTag mice allowed sequencing of actively translated mRNAs and determination of the adult cortical astrocyte transcriptome. Thus, we provide well-characterized, easy-to-use resources with which to selectively study astrocytes in situ and in vivo in multiple experimental scenarios.