Single-cell RNA Sequencing Reveals And Identifies The Main Cell Type Markers In Mice Eyes

The eyes are the organs of the visual system with the ability to receive and process visual details.They detect light signals and convert it into electrochemical impulses in neurons,which are then transmitted to the brain.Several previous studies have revealed the cell types within the retina,but no study has focused on the cell types in the eyeball and their roles in the visual system.In addition,many inherited eye diseases exist and the research and knowledge of these diseases need to be deepened.In this study,we used scRNA sequencing to treat experimental mice.We removed the eye tissues within the sclera of the experimental mice and prepared them into single-cell suspensions for singlecell RNA sequencing.After obtaining the experimental data,we analyzed the genes with higher expression levels in the experimental group and consulted the relevant literature to find out the marker genes that have been reported in different cell types and the richly expressed genes in the eye tissues.Furthermore,we performed frozen sections of the eye tissues within the sclera of the control group mice for antibody verification experiments,and repeated the relevant verification experiments to understand the expression levels of different marker genes in various parts of the eye tissues,so as to find new marker genes of the eye tissues within the sclera,and comprehensively explore the main cell types of the eye tissues within the sclera of mice,and to elucidate the biology of vision and gene expression patterns associated with eye diseases.Through single-cell RNA sequencing,we obtained 1831 single cells with 18,237 expressed genes of the eye tissues within the sclera from the experimental group.In the data analysis,14 different cell populations were found by t-SNE,and we identified 14 corresponding cell types and revealed the main mechanisms of each cell type.Given that the molecular mechanisms of eye diseases are complex and diverse,we also analyzed 546 genes related to human eye diseases,including 35 cataract genes,163 genes of two categories of polygenic retinal diseases,and 348 genes of 31 categories of monogenic retinal diseases.Among these 31 categories of monogenic retinal diseases,we mainly studied 203 genes in 16 categories(a total of seven kinds of diseases)of them.Since TFs play an important role in regulating eye development,we also revealed the expression patterns of 1035 TFs,which are expressed in at least one of the 14 identified cell types.Finally,we found several new marker genes,identified 16 stem cell markers of eyes and concluded that besides M�ller cells,hyalocytes and RPE cells are also highly expressed stem cell markers.Our study will provide rich information for understanding the biological functions of the eyeball and human eye diseases.