Determination of key events in mouse hepatocyte maturation at the single-cell level

Hepatocytes, the liver's predominant cells, perform numerous essential biological functions. However, crucial events and regulators during hepatocyte maturation require in-depth investigation. In this study, we performed single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA sequencing (snRNA-seq) to explore the precise hepatocyte development process in mice. We defined three maturation stages of post-natal hepatocytes, each of which establishes specific metabolic functions and exhibits distinct proliferation rates. Hepatic zonation is gradually formed during hepatocyte maturation. Hepatocytes or their nuclei with distinct ploidies exhibit zonation preferences in distribution and asynchrony in maturation. Moreover, by combining gene regulatory network analysis with in vivo genetic manipulation, we identified critical maturation-and zonation-related transcription factors. This study not only delineates the comprehensive transcriptomic profiles of hepatocyte maturation but also presents a paradigm to identify genes that function in the development of hepatocyte maturation and zonation by combining genetic manipulation and measurement of coordinates in a single-cell developmental trajectory.

Automated summary

In this study, the process of hepatocyte development in mice was investigated using single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA sequencing (snRNA-seq). Three maturation stages of post-natal hepatocytes were defined, each with specific metabolic functions and distinct proliferation rates. Hepatic zonation gradually formed during hepatocyte maturation, with hepatocytes or their nuclei exhibiting preferences in distribution and asynchrony in maturation based on ploidy. Furthermore, critical transcription factors related to maturation and zonation were identified through gene regulatory network analysis and in vivo genetic manipulation.