Identifying Candidate Genes that Underlie Cellular pH Sensitivity in Serotonin Neurons Using Transcriptomics: A Potential Role for Kir5.1 Channels

摘要

FIGURE 1. Methods for FACS collection of dissociated eGFP-expressing (eGFP+) and non-eGFP (eGFP-) medullary raphe neurons. (A) Representative data plotting forward-scatter (FSC) versus side-scatter (SSC) of all recorded sort events (cells). An area (P1) was identified as inclusive of raphe neurons, confirmed in (B) by sorting and plotting the FSC versus NeuN fluorescence of NeuN-labeled (Alexa Fluor 546-tagged) cells (Green events in A and B). (C) Gate establishment for the identification of eGFP+ (5-HT neurons) from ePet-eGFP:SS brainstem tissues by plotting FSC and the fluorescence intensity of Alexa Fluor 488 from collected P1/NeuN+ events, designated eGFP+ (red) or eGFP- (purple). (C’) The distribution of Alexa Fluor 488 intensity versus event counts. (D) qPCR data showing fold-enrichment of 5-HT neuron-specific genes slc6a4 (SERT) and tph2 (tryptophan hydroxylase 2) in eGFP+ (5-HT-enriched) versus non-5-HT neuron-enriched cell pools (P < 0.05; One-way ANOVA).