Single-cell RNA sequencing of zebrafish pancreatic cells during beta-cell regeneration

To better understand the underlying mechanism of beta-cell regeneration in adult zebrafish, we performed single-cell transcriptomic profiling of the pancreatic tissue (using 10X Genomics) at various stages post beta-cell ablation. Overall design: The adult fish (expressing NTR gene under the insulin promoter) were treated with Mtz (Sigma-Aldrich, M3761), dissolved at 10mM concentration in fish water. The animals were treated for 24 hours, protected from light. After 24h, Mtz solution was removed and the animals were placed 3 times in fresh fish water before returning them to standard maintenance conditions. Both untreated and Mtz-treated animals were euthanized by an overdose of tricaine. Samples for each time-point comprised of 6 animals (3 males and 3 females). Single-cell suspension was prepared and 15000 live-cells/per condition were FACS sorted for 10X Genomics.