Single-cell transcriptome analysis of cavernous tissues reveals the key roles of pericytes in diabetic erectile dysfunction
- School of Life Sciences, Gwangju Institute of Science and Technology (GIST), Republic of Korea
- National Research Center for Sexual Medicine and Department of Urolog, Inha University School of Medicine, Republic of Korea
- Program in Biomedical Science & Engineering, Inha University, Republic of Korea
Figures
Figure 1 with 4 supplements
Single-cell transcriptional landscape of mouse cavernous tissues in normal and diabetic conditions.
(A) Schematic workflow of this study. Cavernous tissues from 16-week-old male mice were used for single-cell RNA sequencing (n=5 for each group). DM, diabetes mellitus. (B) Visualization of …
Figure 1—figure supplement 1
Chondrocyte subsets in single-cell RNA sequencing data of mouse cavernous tissues.
(A) Uniform Manifold Approximation and Projection (UMAP) of three chondrocyte clusters in single-cell data of mouse cavernous tissues. (B) Expression of marker genes of three chondrocyte subsets. Myl…
Figure 1—figure supplement 2
Volcano plots showing differentially expressed genes (DEGs) between diabetic and normal conditions in each cell type.
(A) DEGs between diabetic and normal conditions in chondrocytes. (B) DEGs between diabetic and normal conditions in myofibroblast (MFB). (C) DEGs between diabetic and normal conditions in …
Figure 1—figure supplement 3
Gene ontology high in normal compared to diabetes in each cell type.
(A) Gene ontology analysis of the genes higher in normal compared to diabetes in chondrocytes. (B) Gene ontology analysis of the genes higher in normal compared to diabetes in myofibroblast (MFB). (C…
Figure 1—figure supplement 4
Gene ontology high in diabetes compared to normal in each cell type.
(A) Gene ontology analysis of the genes higher in diabetes compared to normal in chondrocytes. (B) Gene ontology analysis of the genes higher in diabetes compared to normal in myofibroblast (MFB). (C…
Figure 2 with 2 supplements
Identification of Lbh as a marker of pericytes.
(A) Expression of well-known marker genes of SMC and pericyte (Cnn1 for SMC, Rgs5 and Pdgfrb for pericyte). (B) Significantly enriched gene sets associated with function of SMC and pericyte. Gene …
Figure 2—figure supplement 1
Pericyte-specifically expressed genes in single-cell RNA sequencing data.
(A) Violin plots showing the expression of known marker genes of pericyte (Cspg4, Kcnj8, Higd1b, Cox4i2) and SMC (Acta2, Myh11, Tagln, Actg2). (B) Violin plots showing the expression of Ednra, Gpc3, …
Figure 2—figure supplement 2
Expressions of Lbh in single-cell RNA sequencing data from tissues other than the penis.
ALH, ascending limb of loop of Henle; PST, proximal straight tubule. (A–D) Violin plots showing the expression of endothelial cell marker (Pecam1), known pericyte markers (Rgs5, Pdgfrb, Cspg4) and Lb…
Figure 3 with 3 supplements
Cell-cell interactions between pericytes and other cell types in normal and diabetes.
(A) CellPhoneDB dot plots showing angiogenesis-associated ligand-receptor interactions from pericytes to other cell types in normal and diabetic samples. p-Values are indicated as circle sizes. The …
Figure 3—figure supplement 1
Cell-cell interactions between pericytes and other cell types showing significant differences between normal and diabetes.
CellPhoneDB dot plots showing ligand-receptor interactions between pericytes and other cell types in normal and diabetes. The plot shows all ligand-receptor interactions with significant differences …
Figure 3—figure supplement 2
Cell-cell communication between cell types in normal and diabetes using CellChat.
(A) The inferred fibroblast growth factor (FGF) signaling network in normal and diabetes. (B) The inferred NT signaling network in normal and diabetes. (C) The inferred PDGF signaling network in …
Figure 3—figure supplement 3
RT-PCR validation of differentially expressed genes from single-cell RNA sequencing analyses in MCPs exposed to NG or HG conditions for 72 hr.
(A) Representative RT-PCR for Lmo2, Junb, Elk1, Hoxd10 in MCPs. (B–E) Normalized band intensity values (n=4). *p<0.05 vs. NG group. The relative ratio of the NG group was arbitrarily set to 1. NG, …
Figure 4 with 2 supplements
LBH improves erectile function under diabetic conditions through induction of neurovascular regeneration.
(A and B) Representative images of immunofluorescence staining of LBH (green)/CD140b (red) in cavernosum tissues and LBH (red) in MCPs under normal and diabetic conditions (in vivo and in vitro). …
Figure 4—figure supplement 1
LBH immunofluorescence staining in corpus cavernosum tissues after infection with lentiviruses containing ORF mouse clone of Lbh.
(A) Immunofluorescence staining of CD31 (green) and LBH (red) in corpus cavernosum tissues treated with lentiviruses ORF control particles (NC) and ORF clone of mouse Lbh (LBH O/E) in diabetic mice. …
Figure 4—figure supplement 2
LBH enhances pericyte angiogenesis and MPG neurite sprouting under high glucose conditions.
(A, B, C, G, and I) TUNEL assay (A, green), immunofluorescence staining of PH3 (B, red), migration (C), tube formation (G) in MCPs, and immunofluorescence staining with neurofilament in MPG tissues …
Figure 5 with 1 supplement
LBH as a marker of pericyte in human corpus cavernosum.
(A) Visualization of single-cell data from human corpus cavernosum using Uniform Manifold Approximation and Projection (UMAP). Each cell type is indicated by a different color. EC, endothelial cells;…
Figure 5—figure supplement 1
Expression of marker genes of pericyte and smooth muscle cells in human penis single-cell RNA sequencing data.
(A) Violin plots showing known pericyte marker genes, CSPG4 and COX4I2. (B) A violin plot showing known SMC marker genes, ACTG2. EC, endothelial cells; FBs, fibroblasts; SMCs, smooth muscle cells; …
Figure 6
LBH-interacting protein identification in mouse cavernous pericytes.
(A) LBH was immunoprecipitated (IP) from whole-MCPs lysates, resolved on SDS-PAGE gels, and stained with Coomassie blue solution. Gel bands indicated by red frame dot line were analyzed by liquid …
Additional files
-
Supplementary file 1
Relative density of mouse angiogenesis array spots.
- https://cdn.elifesciences.org/articles/88942/elife-88942-supp1-v1.xlsx
-
Supplementary file 2
Physiological and metabolic parameters: 2 weeks after treatment with PBS, NC, LBH O/E.
Values are the mean ± SEMs for n=5 animals for each group. STZ, streptozotocin; MSBP, mean systolic blood pressure; NC, lentiviruses ORF control particles; LBH O/E, ORF clone of mouse Lbh lentiviruses, *p<0.05 vs. control group.
- https://cdn.elifesciences.org/articles/88942/elife-88942-supp2-v1.xlsx
-
MDAR checklist
- https://cdn.elifesciences.org/articles/88942/elife-88942-mdarchecklist1-v1.pdf
