Protocol for the purification and transcriptomic analysis of mouse astrocytes using GFAT
Astrocytes are glial cells of the central nervous system that modulate neuronal function. Here, we present glyoxal-fixed astrocyte nuclei transcriptomics (GFAT), a protocol for the purification and transcriptomic analysis of astrocyte nuclei from the cortex and cerebellum of adult and aged fresh mouse brain. We describe steps for tissue dissection, glyoxal fixation, homogenization, nuclei isolation, antibody staining, fluorescence-activated cell sorting, and RT-qPCR or bulk RNA sequencing. GFAT does not require transgenic lines or viral injection and allows parallel astrocyte and neuron profiling. Overall design: Bulk RNA-seq analysis of astrocyte and neuron nuclei in 4 month-old (adult) and 2 year-old (aged) WT C57BL/6 male mice