Journal: Medical Gas Research

Article Title: Mechanisms by which sevoflurane affects cognitive function in aged marmosets and mice: up-regulation of FKBP5 expression in brain microglia

doi: 10.4103/mgr.MEDGASRES-D-24-00155

Figure Lengend Snippet: Identification of microglia in the hippocampus of marmosets after sevoflurane exposure. (A) Visualization of nine major classes of cells via UMAP from snRNA-seq. Each point represents a single cell (droplet barcode) colored by cluster. (B) UMAP visualization of the hippocampus in control and anesthetized marmosets. Green: control group; red: sevoflurane group. (C) Visualization of FKBP5 in the hippocampus of marmosets via UMAP. After anesthesia, FKBP5 was significantly increased in the microglial cluster. (D) Violin plot showing the expression of FKBP5 in nine major clusters of the hippocampus in control and anesthetized marmosets. Ctrl: Control; FKBP5: FK-506 binding protein; OL: oligodendrocytes; OPC: oligodendrocyte precursor cells; Sev: sevoflurane; snRNA-seq: single-cell nuclear RNA sequencing; UMAP: uniform manifold approximation and projection.

Article Snippet: The sections were incubated with rabbit anti-FKBP5 antibody (1:100) and mouse anti-ionized calcium binding adapter molecule 1 antibody (1:500, Servicebio, Wuhan, China, Cat# GB15105, RRID: AB_3663047) at 4°C overnight.

Techniques: Control, Expressing, Binding Assay, RNA Sequencing

Journal: Medical Gas Research

Article Title: Mechanisms by which sevoflurane affects cognitive function in aged marmosets and mice: up-regulation of FKBP5 expression in brain microglia

doi: 10.4103/mgr.MEDGASRES-D-24-00155

Figure Lengend Snippet: Identification of microglia in the hippocampus of mice after sevoflurane exposure. (A) Visualization of nine major classes of cells using UMAP from snRNA-seq. Each point represents a single cell (droplet barcode) colored by cluster. (B) UMAP visualization of the hippocampus in control and anesthetized mice. Green: Control group; red: sevoflurane group. (C) Visualization of FKBP5 in the hippocampus of mice via UMAP. After anesthesia, FKBP5 was significantly increased in the microglial cluster. (D) Violin plot showing the expression of FKBP5 in nine major clusters of the hippocampus between control and anesthetized mice. Ctrl: Control; FKBP5: FK-506 binding protein; Sev: sevoflurane; snRNA-seq: single-cell nuclear RNA sequencing; UMAP: uniform manifold approximation and projection.

Article Snippet: The sections were incubated with rabbit anti-FKBP5 antibody (1:100) and mouse anti-ionized calcium binding adapter molecule 1 antibody (1:500, Servicebio, Wuhan, China, Cat# GB15105, RRID: AB_3663047) at 4°C overnight.

Techniques: Control, Expressing, Binding Assay, RNA Sequencing

Journal: Medical Gas Research

Article Title: Mechanisms by which sevoflurane affects cognitive function in aged marmosets and mice: up-regulation of FKBP5 expression in brain microglia

doi: 10.4103/mgr.MEDGASRES-D-24-00155

Figure Lengend Snippet: The expression of FKBP5 in the hippocampus of mice after sevoflurane exposure. (A) Western blotting was used to validate the expression of FKBP5 in the brains of sevoflurane-treated mice. (B) Quantification of FKBP5 between the control group and sevoflurane group in A ( n = 5, P = 0.0102, t = 3.340). (C) Western blotting validated the expression of FKBP5 in the brains of sevoflurane/surgery-treated mice. (D) Quantification of FKBP5 between the control group and sevoflurane/surgery group in C ( n = 5, P = 0.0002, t = 6.470). The data are presented as mean ± SD. * P < 0.05, *** P < 0.001 (Student’s t -test). Ctrl: Control; Sev 6 h: sevoflurane anesthesia for 6 hours; Sev + Sur 6 h: sevoflurane anesthesia and surgery for 6 hours.

Article Snippet: The sections were incubated with rabbit anti-FKBP5 antibody (1:100) and mouse anti-ionized calcium binding adapter molecule 1 antibody (1:500, Servicebio, Wuhan, China, Cat# GB15105, RRID: AB_3663047) at 4°C overnight.

Techniques: Expressing, Western Blot, Control

Journal: Medical Gas Research

Article Title: Mechanisms by which sevoflurane affects cognitive function in aged marmosets and mice: up-regulation of FKBP5 expression in brain microglia

doi: 10.4103/mgr.MEDGASRES-D-24-00155

Figure Lengend Snippet: Sevoflurane/surgery induces cognitive decline through FKBP5 in microglia. (A) Immunofluorescence staining of the cortex in FKBP5 CKO mice and control mice. FKBP5 was knocked down in microglia of the FKBP5 CKO mouse brain after sevoflurane/surgery. Green: FKBP5, labeled with Alexa Fluor 488; red: IBA-1, a marker of microglia, labeled with CY3; blue: DAPI, labeled with cell nuclei; purple: merged. (B) The latency for male control mice and male FKBP5 CKO mice to identify the target hole in the training phase after exposure to sevoflurane/surgery ( n = 12, P = 0.0006, F (1, 88) = 12.53). (C) The percentage of total time that male control mice and male FKBP5 CKO mice spent in the target hole area during the test phase after exposure to sevoflurane/surgery ( P = 0.0002, t = 4.367). (D) The latency for female control mice and female FKBP5 CKO mice to identify the target hole in the training test after exposure to sevoflurane/surgery ( n = 14, P < 0.0001, F (1, 104) = 18.77). (E) The percentage of total time that female control mice and female FKBP5 CKO mice spent in the target hole area during the test phase after exposure to sevoflurane/surgery ( P = 0.0045, t = 3.110). Heterozygous mice were used as a control group. The data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001 (Student’s t -test (C, E), two-way analysis of variance followed by Sidak’s multiple comparisons test (B, D)). Ctrl: Heterozygous mice; DAPI: 4′,6-diamidino-2′-phenylindole; FKBP5: FK-506 binding protein; FKBP5 CKO: microglia-specific FKBP5 conditional knockout; IBA-1: ionized calcium binding adapter molecule 1.

Article Snippet: The sections were incubated with rabbit anti-FKBP5 antibody (1:100) and mouse anti-ionized calcium binding adapter molecule 1 antibody (1:500, Servicebio, Wuhan, China, Cat# GB15105, RRID: AB_3663047) at 4°C overnight.

Techniques: Immunofluorescence, Staining, Control, Labeling, Marker, Binding Assay, Knock-Out

Journal: Medical Gas Research

Article Title: Mechanisms by which sevoflurane affects cognitive function in aged marmosets and mice: up-regulation of FKBP5 expression in brain microglia

doi: 10.4103/mgr.MEDGASRES-D-24-00155

Figure Lengend Snippet: Transcriptome sequencing analysis of the brains of FKBP5 CKO mice after sevoflurane/surgery. (A) Heatmapmap comparing FKBP5 CKO mice ( n = 10) and control mice ( n = 7) after sevoflurane/surgery. (B) Volcano plot comparing FKBP5 CKO mice ( n = 10) and control mice ( n = 7) after sevoflurane/surgery. The red dots represent RNAs significantly increased in FKBP5 CKO mice compared with control mice (fold change > 1.5, FDR < 0.05); the blue dots represent RNAs significantly decreased in FKBP5 CKO mice compared with control mice (fold change < 0.67, FDR < 0.05). (C–E) GO functional enrichment analysis in terms of biological process (BP) (C), molecular function (MF) (D), and cell composition (CC) (E). (F) KEGG pathway enrichment analysis. Heterozygous mice were used as a control group. Ctrl: Heterozygous mice; FDR: false discovery rate; FKBP5 CKO: microglia-specific FKBP5 conditional knockout.

Article Snippet: The sections were incubated with rabbit anti-FKBP5 antibody (1:100) and mouse anti-ionized calcium binding adapter molecule 1 antibody (1:500, Servicebio, Wuhan, China, Cat# GB15105, RRID: AB_3663047) at 4°C overnight.

Techniques: Sequencing, Control, Functional Assay, Knock-Out