Single cell transcriptome profiling of mouse pancreatic a-lineage cells

As a strategy for regenerative medicine in diabetes, a-to-ß reprogramming, which involves differentiating a cells into ß cells, is an important option. However, while there are many reports on ß cell differentiation, there are insufficient reports on a cell differentiation. There are still many unknowns regarding how a cells mature. In this study, we used the Gcg-Timer mouse model, which enables high temporal resolution to evaluate the development process of a cells. We sorted newly generated and more differentiated a cells from E17.5 Gcg-Timer and performed single-cell RNA sequencing. As a result, we found six clusters within the endocrine population and revealed transcriptional heterogeneity. In addition, the fluorescent characteristics of Gcg-Timer (newly generated and more differentiated cells) did not match the pseudotime information, which was in contrast to the results of our previous analysis of transcriptional characteristics during ß-cell development using Ins1-eGFP;Timer. Furthermore, on the pseudotime axis of Gcg-Timer, Gcg mRNA showed a transient expression pattern, which was different from the expression pattern of Ins1 mRNA in Ins1-eGFP;Timer. These results suggest that (1) a-cell differentiation is probabilistic and (2) glucagon mRNA activation is probabilistic. Thus, evaluating the expression patterns of Gcg mRNA during a-cell development will lead to a deeper understanding of a-cell differentiation in the future. Overall design: This study characterizes the single-cell transcriptomes of mouse pancreatic a-lineage cells.