Single-nuclei sequencing of Parvalbumin interneurons in mouse visual cortex after optoTrkB activation

After deep anesthesia by pentobarbital, the mice were perfused with ice-cold NMDG ACSF, and the brains were isolated and sliced by vibratome in NMDG ACSF. Then the slices were exposed by LED blue light, and incubated in a modified HEPES aCSF for one hour. The visual cortex was Isolated from the slices under microscope with red light, and tissues were homogenized, and nuclei were extracted. All procedures were performed with RNase free in the dark condition. The nuclei were stained with Hoechst 33342 and mouse monoclonal anti-NeuN antibody (Sigma-Aldrich, MAB377), and double positive nuclei were sorted by FACS. Then the sorted nuclei were used to make cDNA Library with Chromium Single Cell 3ʹ Gene Expression v3 library preparation kit (10x Genomics), and sequenced with NovaSeq 6000 (Illumina). Data was pre-processed using CellRanger 3.0, and analysed by Chipster version 3.16/Seurat v3. snRNA-seq of neurons in the visual cortex