The Single Cell Transcriptome Study Of Apoe^-/- Mice And Ldlr^-/- Mice Retina

Abnormal lipid metabolism can cause many diseases and seriously endanger human health.Abnormal lipid metabolism is related to many eye diseases.In order to alleviate or treat retinal physiological changes caused by abnormal lipid metabolism,we usesingle-cell transcriptome sequencing technology to study the retinal tissue of Apoe gene-deficient mices and Ldlr gene-deficient mices,which is to excavate differential expression gene,signal pathway,and carry out functional research to provide theoretical basis for the treatment of retinal lipid metabolism disorders.The main research content is divided into three parts.The single cell transcriptome of the mouse retinas were sequenced.Mices were induced by treating high-fat diet to exacerbate symptoms of abnormal lipid metabolism.Retinas were stripped and prepared into single-cell suspensions for detecting and sequencing.After sorting and analyzing the sequencing results,we summarized the cell types and the differentially expressed genes of each cell type in Apoe^-/-mice and Ldlr^-/-mice by comparing with wild-type mice.The differentially-expressed genes were sorted out and screened in Apoe^-/-mice and Ldlr^-/-mice which had up regulated differently expressed genes.In eventually,we found that Rtn4 was up regulating differentially expressed gene in both Apoe^-/-mice and Ldlr^-/-mice,mainly expressed in Müller cells.Combined with literatures,Rtn4 is a nerve regeneration inhibitory factor which can inhibit the repair function of nerve cells after injury.Then,by adding LDL or NEP1-40 to treat the human Müller cells in order to measure the expression fo Rtn4 by real-time luorescence quantitative PCR and the immunocytochemical experimental technology.It found that the expression of RTN4had increased after LDL treatment of Müller cells;the expression of RTN4had be inhibited after a certain amount of NEP1-40 treatment of Müllercells.RTN4 was mainly expressed in Müller's cytoplasm.Its expression pattern was similar to cytoskeleton proteins,and it was more strongly expressed in pseudopods and intercellular junctions.In summary,Rtn4 is mainly expressed in Müller cells of the retina,and its regrowth inhibiting function can be inhibited by the antagonist NEP1-40.NEP1-40treat the vitreous of mices can increase the thickness of the outer nuclear layer of mice retina.The expression pattern of Rtn4 in the retina may be affect by Apoe.The expression of RTN4 increased after LDL treatment of Müller cells,and a certain amount of NEP1-40 treatment can inhibited the expressio.Combining all results,we find that the lipid transport pathway of APOE\LDL\LDLR may regulate the expression or functional of RTN4,which may cause abnormal lipid metabolism pathways and related eye diseases.The use of NEP1-40 antagonists may inhibit or alleviate eye diseases which caused by abnormal LDLR gene and APOE gene.