Meeting Abstract
RNAseq provides the ability to fully analyze comprehensively changes in gene expression. Our RNAseq studies on the small intestine of the Burmese python revealed approximately 1800 genes that are differentially expressed with the onset of digestion. For the python intestine, genes were equally downregulated or upregulated in expression with feeding, with expression patterns reversed upon the completion of digestion. With the aim of matching gene expression patterns to phenotypic changes of the intestine, specifically the structural and functional changes of enterocytes, we must however consider that the intestinal mucosa is composed of different cell types (e.g., enterocytes, smooth muscle, endocrine, immune, etc.), and that our tissue expression data reflects the combined expression pattern of each cell type. In order to generate cell-specific transcriptional data, we employed single-cell RNAseq on intestinal tissue from a 4-day postfed boa constrictor (Boa constrictor). Using the Fluidigm C1 platform, we constructed cDNA libraries from 54 live cells. Illumina RNAseq libraries created from these cDNA were pooled and sequenced. Analysis of single-cell gene expression enabled us to differentiate among, and to identify, distinct cell types of the boa’s intestine. We therefore are able to identify divergent gene expression patterns across cell types, to link differential expression signatures to distinct cellular phenotypic responses, and to develop cell-specific signaling pathways.
