Study On Cells Heterogeneity Of Rat Cerebral Cortex Based On High-throughput Single Cell Chromatin Accessibility Sequencing Technology

Background and ObjectivesCell theory provides a new framework for understanding biology and disease by asserting that cells are the basic unit of life.Then it was discovered that DNA is a genetic program that encodes proteins that perform cell functions.This discovery led to the development of modern genetics and genomics.The Human Genome Project results show that DNA codes are transformed into functions depending not only on protein-coding sequences but also on gene expression regulatory elements in nonprotein-coding regions.In the brain's central nervous system,people have been lacked a comprehensive catalog of their gene expression.The construction of the brain's apparent regulatory network is essential for understanding the of the brain's complex functions.At present,the research on the epigenetic regulatory network at the population level is very mature.However,the population level is often the mean value of gene expression or represents the dominant cell life activity information.The biological differences between cells may be averaged or be covered up by mistakenly thinking of technical noise.At this stage,some single-cell epigenomics sequencing technologies have emerged.Due to the low throughput of these technologies and the low coverage of the whole genome,the available single cells and the information captured by every single cell are limited,which gives large-scale the precise definition of the single-cell state presents obstacles.Recently mature single-cell RNA omics technology has been used to reveal cells' diversity and specificity in the cerebral cortex.However,the development of single-cell chromatin access technology has many challenges in data quality,throughput,cost,and operability.Methods1.The first part of this project developed a high-throughput single-cell chromatin accessibility technology.The high-throughput single-cell capture rate provides a basis for the large-scale study of the chromatin open area of species and good data quality,which performed.Operability accelerates chromatin accessibility maps' construction and provides public resources for in-depth analysis of gene expression regulation mechanisms.2.Based on human brain samples' preciousness and the rat brain is a better model for studying human brains than mouse brains.We are the first to use this technology to construct a chromatin accessibility map of the brain's essential cortical regions.ResultsBy obtaining 59,023 single cells,we systematically described the single-cell chromatin access map in the four cerebral cortex regions of adult Sprague Dawley(SD)rats.We identified the cell types in the cerebral cortex and found that different cortical regions showed diversity in cell composition and gene regulatory regions.And further analysis of specific transcription factors related to diseases,several cell type-specific transcription factors(TFs),including SPI1,KLF4,KLF6,and NEUROD2,have been shown to play an essential role in the pathogenesis of various neurological diseases,such as Alzheimer's disease(AD),astroglioma,autism spectrum disorder(ASD)and intellectual disability.ConclusionsIn this project,we use the developed single-cell ATAC sequencing(single-cell assay for transposase-accessible chromatin using sequencing,scATAC-seq)technology to study the rat cortex and correlate gene expression regulatory elements and gene expression.It's a valuable resource to analyze the regulatory mechanism of the fate of different cortical cells.And further,reveal the regulatory network of neuropathogenesis.The new technology developed provides essential research tools for epigenome research.