Advancements in Single-Cell Profiling in Human Brain Development

The study titled, "Single-cell spatiotemporal transcriptomic and chromatin accessibility profiling in developing postnatal human and macaque prefrontal cortex," was conducted by researchers at Peking University First Hospital and Beijing Normal University.

The study involved 14 female and 23 male human samples for single-nuclei RNA sequencing, four female and 10 male samples for single-nuclei ATAC sequencing, and six female and seven male samples for spatial transcriptomic sequencing.

Ethical approvals were obtained from the Medical Ethics Committee of Peking University First Hospital and the Ethics Committee of Beijing Normal University, with informed consent secured from children's parents and legal guardians for the use of surgical samples.

In addition, the research utilized crab-eating macaques provided by Xieerxin Biology Resource, with animal care procedures approved by the Institutional Animal Care and Use Committee of Beijing Normal University.

The human and macaque tissues were processed for experiments, including freezing in liquid nitrogen or fixing in paraformaldehyde for various analyses. The study employed sophisticated methodologies including single-nuclei RNA sequencing, single-nuclei ATAC sequencing, and spatial transcriptomic sequencing to profile cellular dynamics in the developing prefrontal cortex.

Researchers isolated nuclei from the tissue samples and prepared them for sequencing, utilizing 10x Genomics platforms to generate single-cell libraries. The sequencing data was then analyzed, focusing on integrating datasets across species to identify cellular and molecular differences in brain development.

Key findings highlighted specific gene expression patterns and cell type annotations, revealing distinct clusters of neurons and glial cells in both human and macaque samples. The researchers also identified differentially expressed genes and conducted Gene Ontology enrichment analyses to understand functional implications.

Pseudotime trajectory analysis was utilized to trace developmental pathways of oligodendrocyte precursor cells and astrocytes. The implications of these findings could enhance the understanding of neurological disorders and brain development mechanisms, as noted in the report which emphasizes the relevance of single-cell transcriptomic profiling in advancing neuroscience research.