Fig 1: Hypocretin‐1 induced the decrease of lactate and glycolysis rate in hippocampal astrocytes. a) Schematic diagram showing the detection of glycolytic analysis, lactate, RT‐qPCR and immunofluorescent staining. SB‐334867, a HCRTR1 antagonist. b) The co‐expression of HCRTR1 and GFAP. c) The lactate in astrocyte medium in different groups. CTR, control; HCRT‐1, hypocretin‐1; SB‐334867, HCRT‐1 + SB‐334867. d) Changes of mRNA expression of glycolytic‐related factors in astrocytes. *Compared with CTR, #compared with HCRT‐1. e–g) Glycolysis rate test. The basal, compensatory glycolysis rate and extracellular acidification rate (ECAR) of astrocytes, respectively. h) Quantification of mean fluorescence intensity of the HIF‐1α expression in astrocytes. i) Representative microscopic images showing expression of HIF‐1α in astrocytes. j) Schematic diagram showing treatment with HIF‐1α agonist. k) Changes of mRNA expression of glycolytic‐related factors in astrocytes treatment with HCRT‐1 and HIF‐1α agonist. Dots in panels represent individual samples. Data were shown as mean ± SD. One‐way ANOVA followed by Dunnett's post hoc test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001; ns, no significant difference, p ≥ 0.05.
Fig 2: Hypocretin‐1 induced anxiety, depressive‐behaviors, cognitive impairment and dysregulation of energy metabolism. a) Schematic illustration of the intraventricular injection hypocretin‐1 (icv.HCRT‐1) in hippocampus and construction of CUMS‐induced depression model. b) Both icv.HCRT‐1 and CUMS resulted in decreased center time in open field test. c) Both icv.HCRT‐1 and CUMS decreased open arm time in elevated plus test. d) Both icv.HCRT‐1 and CUMS resulted in decreased new zone exploration time in Y‐maze test. e) Both icv.HCRT‐1 and CUMS resulted in decreased sucrose preference. f) Both icv.HCRT‐1 and CUMS resulted in increased immobility time in forced swim test. g) Representative microscopic of PSD‐95 and SYP in hippocampus. scale bar, 200 µm. h) Representative microscopic of DCX positive cells in the DG. scale bar, 100 µm. i,j) Quantification of mean fluorescence intensity SYP and PSD‐95, respectively. k) Quantification of DCX positive cells in the DG. l) mRNA expression of synapse‐related gene (SYP and PSD‐95) in hippocampus. m) Changes of mRNA expression of glycolytic‐related factors in hippocampus of different groups. n) Hippocampal lactate level in different groups. o) Representative 18F‐FDG PET images of rat hippocampus from control (CTR), icv.HCRT‐1 and CUMS groups. p) Glucose metabolism in hippocampus. Dots in panels represent individual samples. Data were shown as mean ± SD. One‐way ANOVA followed by Dunnett's post hoc test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, no significant difference, p ≥ 0.05.
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