Spinal cord injury (SCI) represents a complex pathophysiological process involving the interaction of multiple cell types. Conventional sequencing methods can only detect the average gene expression level of the damaged local cell populations, which is difficult to reflect its heterogeneity. Therefore, new technologies are needed to reveal the intercellular heterogeneity and the complex intercellular interactions of the damaged lesions. The single-cell RNA sequencing (scRNA-seq) technique facilitates high-resolution profiling of gene expression at the single-cell level, providing insights into cellular heterogeneity and function, potential molecular pathways, cell fate transitions, and the intercellular interactions pertinent to disease progression. This technology generates valuable gene expression data that support both basic and translational research efforts aiming at the identification of therapeutic targets for intervention. The scRNA-seq technique and its multifaceted application in the local immune microenvironment of injury after SCI were discussed, which will contribute to a more comprehensive understanding of the pathophysiological processes in the immune microenvironment of SCI.
Spinal Cord Injuries/genetics* ; Humans ; Single-Cell Analysis/methods* ; Sequence Analysis, RNA/methods* ; Animals ; Gene Expression Profiling/methods* ; Cellular Microenvironment/genetics*

Objective To observe the expression characteristics of green fluorescent protein(GFP)-positive myeloid cells(mainly monocytes,macrophages,and neutrophils)and their pattern of response to diphtheria toxin(DT)in Lyz2 internal ribosome entry site(IRES)DT receptor/enhanced GFP(DTREGFP)mice.Methods Transgenic Lyz2 IRES DTREGFP mice(6～8 weeks old)and C57BL/6J mice with the same genetic background were selected randomly and injected intraperitoneally with DT(25 ng/g)for 3 consecutive days.Bone marrow,peripheral blood,and spleen cells were separated and made into single-cell suspensions with GFP and CD11b as markers.Changes in the proportion of GFP+cells to total myeloid cells(CD11b+)was detected in these tissues at different time points before and after drug administration,using flow cytometry.Results Peripheral blood CD11b+GFP+cells accounted for(24.62±5.655)％of CD11b+cells before DT injection,and this proportion gradually decreased after injection and reached a minimum of(7.982±2.729)％on the third day.The proportions in spleen and bone marrow were(13.73±2.994)％and(65.23±4.261)％before DT injection,respectively,decreased most significantly on the first day after injection to(3.468±0.5862)％and(50.98±7.957)％,respectively,and then gradually recovered.Conclusions DT effectively reduced the proportions of monocytes,macrophages,and neutrophils in Lyz2 IRES DTREGFP mice,but the pattern of change varied among tissues.

Objective To observe the expression characteristics of green fluorescent protein(GFP)-positive myeloid cells(mainly monocytes,macrophages,and neutrophils)and their pattern of response to diphtheria toxin(DT)in Lyz2 internal ribosome entry site(IRES)DT receptor/enhanced GFP(DTREGFP)mice.Methods Transgenic Lyz2 IRES DTREGFP mice(6～8 weeks old)and C57BL/6J mice with the same genetic background were selected randomly and injected intraperitoneally with DT(25 ng/g)for 3 consecutive days.Bone marrow,peripheral blood,and spleen cells were separated and made into single-cell suspensions with GFP and CD11b as markers.Changes in the proportion of GFP+cells to total myeloid cells(CD11b+)was detected in these tissues at different time points before and after drug administration,using flow cytometry.Results Peripheral blood CD11b+GFP+cells accounted for(24.62±5.655)％of CD11b+cells before DT injection,and this proportion gradually decreased after injection and reached a minimum of(7.982±2.729)％on the third day.The proportions in spleen and bone marrow were(13.73±2.994)％and(65.23±4.261)％before DT injection,respectively,decreased most significantly on the first day after injection to(3.468±0.5862)％and(50.98±7.957)％,respectively,and then gradually recovered.Conclusions DT effectively reduced the proportions of monocytes,macrophages,and neutrophils in Lyz2 IRES DTREGFP mice,but the pattern of change varied among tissues.