Fig. 7

GLUT1-specific knockdown in hippocampal microglia alleviates microglial proinflammatory state and CUMS-related spatial learning and memory impairment. (A) Schematics of the AAV construct expressing GLUT1-targeted microRNAs specifically in microglia (AAV-mirGLUT1) (upper). AAV-mirGLUT1 or AAV-control was injected into the mouse hippocampus, and the experimental timeline is shown (lower). (B) Representative fluorescence image of GFP in the mouse hippocampus after AAV-mirGLUT1 infection. Scale bar, 100 μm. (C) qRT-PCR assay monitoring the expression of GLUT1 in the mouse hippocampus injected with AAV-mirGLUT1 or AAV-control (n = 6, Student’s t-test). (D&E) qRT-PCR assays monitoring expression levels of proinflammatory phenotype markers (D) and anti-inflammatory phenotype markers (E) in hippocampal samples from CTRL and CUMS mice infected with AAV-mirGLUT1 or AAV-control (n = 6, One-way ANOVA with Tukey’s post hoc test). (F) Levels of IL-6, IL-1β, and TNF-α in hippocampus lysates from CTRL and CUMS mice infected with AAV-mirGLUT1 or AAV-control as determined by ELISA (n = 6, One-way ANOVA with Tukey’s post hoc test). (G) Escape latency to the platform during the training trials in the MWM test of CTRL and CUMS mice injected with AAV-mirGLUT1 or AAV-control (n = 4, Two-way ANOVA with Tukey’s post hoc test). (H-J) Latency to enter the platform (H), swimming distance of first time to enter the platform (I), and platform crossings (J) in the probe trial of the MWM test of CTRL and CUMS mice after AAV-mirGLUT1 or AAV-control infection (n = 4, One-way ANOVA with Tukey’s post hoc test). *p < 0.05, **p < 0.01