Journal: Neural Regeneration Research

Article Title: Gamma-glutamyl transferase 5 overexpression in cerebrovascular endothelial cells improves brain pathology, cognition, and behavior in APP/PS1 mice

doi: 10.4103/NRR.NRR-D-23-01525

Figure Lengend Snippet: GGT5 expression is downregulated in Aβ 1–42 –treated hCMEC/D3 and bEnd.3 cells and in the brains of APP/PS1 mice. (A) mRNA-seq analysis of hCMEC/D3 cells identified 178 genes that were differentially expressed between the vehicle and Aβ 1–42 intervention groups. (B) Nine downregulated genes and two upregulated genes were selected by comparing the 178 differentially expressed genes with an existing AD single-cell sequencing database. (C) qPCR verification of the expression levels of the 11 selected genes ( n = 3 per group). (D, E) Representative western blots (D) and quantitative analysis (E) of AGRN, LRP1, and GGT5 expression in hCMEC/D3 cells treated with vehicle or Aβ 1–42 (each row is an image from a different blot) ( n = 3–4 per group). (F, G) Representative western blots (F) and quantitative analysis (G) of GGT5 expression in bEnd.3 cells treated with vehicle or Aβ 1–42 ( n = 4 per group). (H–K) Representative western blots of GGT5 and Aβ in the Ctx (H) and Hip (I) and quantitative analysis of GGT5 (J) and Aβ (K) expression in the Ctx and Hip of mice ( n = 4 per group). Data are expressed as mean ± SEM. * P < 0.05, *** P < 0.001 (one-way analysis of variance followed by Tukey’s post hoc test [J and K], or independent-samples t -test [C, E and G]). AD: Alzheimer’s disease; AGRN: Agrin; ARHGEF17: Rho guanine nucleotide exchange factor (GEF) 17; Aβ: amyloid-β; Ctx: cortex; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GGT5: gamma-glutamyltransferase 5; Hip: hippocampus; LRP1: low density lipoprotein receptor related protein 1; MCAM: melanoma cell adhesion molecule; MRC2: mannose receptor c type 2; PLOD1: procollagen lysine-1,2-oxoglutarate-5-dioxygenase 1; RPS27A: ribosomal protein S27A; WT: wide type.

Article Snippet: One milligram of recombinant human Aβ 1−42 peptide (ChinaPeptides, Shanghai, China) was dissolved in 60 μL of dimethyl sulfoxide (DMSO) (Li et al., 2022b) and then diluted to the final concentration (5 μM) with complete medium.

Techniques: Expressing, Sequencing, Western Blot

Journal: Neural Regeneration Research

Article Title: Gamma-glutamyl transferase 5 overexpression in cerebrovascular endothelial cells improves brain pathology, cognition, and behavior in APP/PS1 mice

doi: 10.4103/NRR.NRR-D-23-01525

Figure Lengend Snippet: GGT5 overexpression in cerebrovascular endothelial cells reduces the Aβ load in the brains of APP/PS1 mice by inhibiting BACE1. (A–F) Representative western blots (each row is an image from a different blot) (A) and quantitative analysis of flAPP (B), CTF (C), BACE1 (D), total Aβ (E), and Aβ 1–42 (F) expression in the mouse cortex ( n = 3–5 per group). (G–I) Representative western blots (each row is an image from a different blot) (G) and quantitative analysis of BACE1 (H) and total Aβ (I) expression in the mouse hippocampus ( n = 4–5 per group). Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s post hoc test). APP: Amyloid precursor protein; Aβ: amyloid-β; BACE1: β-site APP cleaving enzyme1; CTF: C-terminal fragment; Ctx: cortex; flAPP: full-length APP; GGT5: gamma-glutamyltransferase 5; Hip: hippocampus; WT: wide type.

Article Snippet: One milligram of recombinant human Aβ 1−42 peptide (ChinaPeptides, Shanghai, China) was dissolved in 60 μL of dimethyl sulfoxide (DMSO) (Li et al., 2022b) and then diluted to the final concentration (5 μM) with complete medium.

Techniques: Over Expression, Western Blot, Expressing