Human islets exposed to conditions that induce or inhibit calcium signaling

Pancreatic islets depend on cytosolic calcium to trigger the secretion of glucoregulatory hormones and the transcriptional regulation of genes important for islet response to stimuli. To date, there has not been an attempt to profile calcium-regulated gene expression in all islet cell types. To address this, we generated a large single cell transcriptomic dataset from healthy human islets exposed to conditions that would acutely induce or inhibit intracellular calcium signaling, while preserving biological heterogeneity. Our aim was to use this dataset to identify acutely calcium-regulated genes in each islet cell type, while simulataneously exploring markers of islet heterogeneity. Islets were obtained from 3 healthy, age-matched, BMI-matched, male human donors: R253, R282, and R317. Each islet prep was split into 3 experimental conditions designed to induce or inhibit calcium signalling pathways: Low, Positive, and Negative. Each experimental condition within each donor prep was handled as an individual "sample" throughout the library preparation and analysis, resulting in 9 samples composed of 3 biological replicates of 3 experimental conditions. For sequencing, all libraries generated from the 3 experimental conditions were pooled per donor, and sequenced on a single flow cell. For each donor's 3 libraries, sequencing was performed twice in order to meet an appropriate number of reads, resulting in 2 technical replicates of sequencing: run1 and run2. In total, there were 18 individual samples' sequencing data carried downstream for analysis, composed of the 9 individual samples, each sequenced across 2 sequencing runs.