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. Author manuscript; available in PMC: 2023 Jul 1.
Published in final edited form as: Biol Psychiatry. 2022 Feb 18;92(1):68–80. doi: 10.1016/j.biopsych.2022.02.007

Figure 1. Aldh1l1-eGFP/Rpl10a mice allow for isolation of astrocyte-specific ribo-associated mRNA.

Figure 1.

Schematic illustrating the immunoprecipitation (IP) of astrocyte-specific ribo-associated mRNA from whole NAc tissue of Aldh1l1-eGFP/Rpl10a mice that express an eGFP tag on the ribosomal subunit RpL10a. (B) Isolated mRNA is highly specific for astrocytes, as revealed by a cell-type enrichment heatmap generated from plotting Log2CPM values of established cell-type markers from our RNA-sequencing dataset (alternate names: Slc1a2 = EAAT2 or Glt-1; Slc1a3 = EAAT1 or Glast; Glul = GS; Rbfox3 = NeuN; Slc17a6 = Vglut2). Columns are the individual samples, rows are individual genes. Purple indicates low expression, while yelllow indicates peak expression. (C) Moreover, the fluorophore eGFP is expressed specifically in cells expressing the pan-astrocyte marker Aldh1l1, but not in cells expressing the neuronal GABA transporter VGAT, as determined by RNAscope; arrows indicate cells with co-expression of Aldh1l1 and eGFP. (D) Following RNA-seq, Ingenuity Pathway Analysis (62) and Metascape analysis (63) revealed the top canonical pathways and biological processes enriched among the top 200 expressed genes in NAc astrocytes include key metabolic and synaptic neurotransmission functions generally attributed to astrocytes. Notable NAc-astrocyte enriched processes include glycolysis and lactate synthesis, oxidative phosphorylation, glutamate receptor signaling, neurotransmitter uptake (e.g., glutamate), and neuroinflammation signaling. Red line in bar graphs indicates significance threshold of p<0.05 or −log10(p-value) > 1.3.