Transcriptomic and neurochemical analysis of the stellate ganglia in mice highlights sex differences

Study Purpose: The stellate ganglia are the predominant source of sympathetic innervation to the heart. Remodeling of the nerves projecting to the heart has been observed in several cardiovascular diseases, however, studies of adult stellate ganglia are limited. A profile of the baseline transcriptomic and neurochemical characteristics of the stellate ganglia in adult C57Bl6J mice, a common model for the study of cardiovascular diseases, may aid future investigations.

Data Collection: RNA-sequencing was used to identify expression differences between stellate ganglia in male and female mice. Six biological replicates for female and male stellate ganglia samples were contrasted by high-throughput sequencing using the Illumina HiSeq 2500. The analysis contains differential expression analysis and gene ontology analysis of these differences, including physiologically important genes for growth factors, receptors, and ion channels. Neurochemical analysis was also performed on stellate ganglia and heart tissue samples.

Primary Conclusion: While the neurochemical profiles of male and female stellate ganglia were not different, minor differences in neurotransmitter content were identified in heart tissue. RNA-sequencing analysis revealed significant sex differences in expression of genes encoding growth factor receptors (TrkC, Ret), ion channels (Kv1.2, KCa3.1), vesicular monoamine transporter 1 (VMAT1), and neuronal guidance molecules (Netrin G1).

Curator's Notes

Experimental Design: This dataset contains derivative analyses of data originally published in Bayles et al. (2018), DOI: 10.1038/s41598-018-27306-3. Raw RNA-sequencing data (fastq files) are publicly available at NCBI BioProject PRJNA423086 with individual sample accession numbers SRR6487140-SRR6487153. Briefly, left stellate ganglia were collected from male (n=6) and female (n=6) C57BL/6J mice aged 16-19 weeks. RNA was extracted using the RNAqueous Micro kit (Ambion), and sequencing libraries were prepared using the TruSeq Stranded Total RNA Library Prep Kit with RiboZero ribosomal RNA removal (Illumina) with 130 ng RNA per sample. Sequencing was performed on an Illumina HiSeq 2500 at Oregon Health & Science University. The derivative data in this dataset includes processed gene expression data normalized using the trimmed mean of M-values method (TMM), differential expression analysis results between sexes performed using limma with empirical Bayes moderation and false discovery rate (FDR) adjustment, and gene ontology analysis. Neurochemical measurements of norepinephrine, acetylcholine, dopamine, and metabolites were performed on separate cohorts using HPLC and mass spectrometry as described in the original publication. This dataset provides tabularized summaries of these data.

Completeness: This dataset is complete.

Subjects & Samples: Female (n=12), male (n=12) C57BL/6J mice (RRID:IMSR_JAX:000664) age 16-19 weeks were used in this study. Six mice of each sex were used for RNA-sequencing (expression group) and six mice of each sex were used for neurochemical analysis (neurochemistry group).

Primary vs derivative data:. There is no primary data included in this dataset. Original raw fastq sequencing data are available online at NCBI BioProject PRJNA423086. Derivative data includes processed figures (.tiff) showing expression levels, multidimensional scaling plots, and qPCR validation; tables (Word format) with cardiac and stellate ganglion neurochemistry data and differentially expressed genes; excel files containing raw count and normalized CPM expression data, statistical comparisons between sexes, top 100 expressed genes, gene class analyses, neuronal subpopulation data, and individual mouse neurochemistry values; and a DiffExp Gender pdf document with quality control metrics, sequence alignment details, and differential expression analysis. This analyzed dataset is derived from data originally published at https://doi.org/10.1038/s41598-018-27306-3.