Desciption of the transcriptome profiles of mouse heart innervating DRG and Nodose Ganglia-derived sensory neurons

To obtain the mRNA expression profiles of vagal and spinal cardiac sensory neurons, mouse Dorsal Root Ganglia (DRG) and Nodose Ganglia (NG) neurons innervating the heart were labelled by retrograde Di-8-ANEPPQ dye which was injected to the heart. Then bilateral NGs and DRGs between C-T segments were dissected and Di-8-ANEPPQ-labelled DRG and NG neurons were FACS sorted. Finally, Di-8-ANEPPQ+ DRG and NG neurons were subjected to total RNA extraction and library preparation protocols for RNA-seq experiment. For control samples, total NG and DRG (C-T) neurons were dissected from the wt mice and same RNA extraction and library preparation protocols were applied. Finally, mRNA profiles of heart-spesific DRG and NG neurons were compared to those of total DRG and NG populations, respectively. All samples were obtained from female Balb-C mice at 8-12 weeks. The following samples were used for RNA-sequence analysis: The heart-specific DRG (DRG-HS, three samples, each sample was obtained from three or four animals (n=3/4)), the heart-specific NG (NG-HS; two samples, each sample was obtained from two animals), sham DRG (DRG-T; two samples, each sample was obtained from one animal (n=1)), and sham NG (NG-T; two samples, each sample was obtained from one animal (n=2)). By using the RNeasy micro kit (Qiagen), RNA was extracted from each sample. Before further progress, RNA concentrations were measured using Qubit (Thermo Fisher Scientific) and RNA integrities were assessed on Agilent 2100 Bioanalyzer. In RNA Seq, only the samples with the an average of 7.5 RNA integrity number (RIN) were included. RNA libraries were prepared using Truseq Total RNA Sample Preparation Kit v2. 500 ng/µl and 200 ng/µl of RNA were used for each DRG and NG sample, respectively. Fragments were sequenced as paired-end reads (2 x 75 bp) on the Illumina NextSeq 500 platform in İstanbul Medipol University, Center of Genomics.