Journal: Cell Insight

Article Title: The role of TRPV4 in acute sleep deprivation-induced memory impairment: Mechanisms of calcium dysregulation and synaptic plasticity disruption

doi: 10.1016/j.cellin.2025.100240

Figure Lengend Snippet: Characterization of differentially expressed genes (DEGs) between the control and ASD groups. (A) Timeline of behavioral modeling and mRNA sequencing. (B) Volcano plots showing upregulated (yellow) and downregulated (blue) DEGs in ASD vs. Control (Ctr) in the PL. (C) Heatmap of DEG based on mRNA sequencing data from the two groups ( p < 0.05). (D). Gene Ontology (GO) term analysis of ASD FC vs. Ctr FC, n = 3 per group, p value ​< ​0.05. (E)-(F) Verification of upregulated genes (Trpv4, Pim1, and Htr5b) and downregulated genes (Adnp, Dbp, and Henmt1) using RT-qPCR in two groups. FC: fear conditioned; ASD: acute sleep deprivation; Mean ±SEM. ∗ p ​< ​0.05, ∗∗ p < 0.01.

Article Snippet: Control lentivirus and TRPV4 shRNA lentivirus (approximately 1 μL at a rate of 80 nL/min) were injected bilaterally into the PL (2.34 mm anterior to the bregma, 0.20 mm lateral to the midline, and 1.90 mm ventral to the dura) using a glass pipette connected to a microinjection pump (RWD Life Science, Shenzhen, China).

Techniques: Control, Sequencing, Quantitative RT-PCR

Journal: Cell Insight

Article Title: The role of TRPV4 in acute sleep deprivation-induced memory impairment: Mechanisms of calcium dysregulation and synaptic plasticity disruption

doi: 10.1016/j.cellin.2025.100240

Figure Lengend Snippet: TRPV4 expression is increased in the PL following acute sleep deprivation but is not influenced by fear conditioning. ( A) Acute sleep deprivation significantly increased TRPV4 mRNA expression in both the context control and fear-conditioned groups ( n ​= ​5–6 per group). (B) TRPV4 protein expression was significantly higher in the ASD FC group compared to the Ctr FC group ( n ​= ​3 per group). FC: fear conditioned; ASD: acute sleep deprivation; Mean ±SEM; ∗∗∗ p ​< ​0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Control lentivirus and TRPV4 shRNA lentivirus (approximately 1 μL at a rate of 80 nL/min) were injected bilaterally into the PL (2.34 mm anterior to the bregma, 0.20 mm lateral to the midline, and 1.90 mm ventral to the dura) using a glass pipette connected to a microinjection pump (RWD Life Science, Shenzhen, China).

Techniques: Expressing, Control

Journal: Cell Insight

Article Title: The role of TRPV4 in acute sleep deprivation-induced memory impairment: Mechanisms of calcium dysregulation and synaptic plasticity disruption

doi: 10.1016/j.cellin.2025.100240

Figure Lengend Snippet: Knockdown of TRPV4 in the PL reversed memory impairment induced by sleep deprivation. (A) Fluorescent images showing the location of the TRPV4-shRNA lentivirus infusion. Scale bars: 100 μm (white) for the main image and 10 μm (red) for the inset. (B) The knockdown efficiency of TRPV4-shRNA was approximately 50 ​%. (C) Timeline of lentivirus infusion, sleep deprivation, and behavioral testing. (D) TRPV4-shRNA lentivirus did not affect memory acquisition. (E) The percentage of freezing was significantly higher in animals infused with TRPV4-shRNA compared to those infused with the control virus after sleep deprivation. (F) Timeline of sleep deprivation, behavioral training, and lentivirus infusion. (G) Sleep deprivation did not affect memory acquisition. (H) Freezing levels were significantly higher in ASD animals infused with TRPV4-shRNA compared to the control virus. FC: fear conditioned; ASD: acute sleep deprivation; Mean ±SEM. ∗ p < 0.05.

Article Snippet: Control lentivirus and TRPV4 shRNA lentivirus (approximately 1 μL at a rate of 80 nL/min) were injected bilaterally into the PL (2.34 mm anterior to the bregma, 0.20 mm lateral to the midline, and 1.90 mm ventral to the dura) using a glass pipette connected to a microinjection pump (RWD Life Science, Shenzhen, China).

Techniques: Knockdown, shRNA, Control, Virus

Journal: Cell Insight

Article Title: The role of TRPV4 in acute sleep deprivation-induced memory impairment: Mechanisms of calcium dysregulation and synaptic plasticity disruption

doi: 10.1016/j.cellin.2025.100240

Figure Lengend Snippet: Ca 2+ concentration was reduced, and impaired synaptic plasticity was reversed in the PL following TRPV4-shRNA lentivirus infusion in sleep-deprived mice. (A) Timeline of tissue collection for measuring Ca 2+ concentration and PSD95 expression after TRPV4-shRNA knockdown. (B) TRPV4 knockdown significantly reduced the concentration of Ca 2+ in the PL of sleep-deprived mice. (C) PSD95 mRNA level was significantly increased in the ASD TRPV4-shRNA group. (D) Representative immunoblot images of PSD95 in the two groups. (E) PSD95 protein expression was significantly increased after TRPV4-shRNA infusion compared to scramble virus infusion. (F) Representative immunofluorescence of PSD95 in the two groups. (G) Quantitative analysis showed that PSD95 protein expression was significantly enhanced. (H) Timeline for dendrite spine density analysis in different groups. (I) Schematic diagram showing dendritic spines of PL neurons. Secondary basal spine dendrites were analyzed. Scale bar: 10 μm. (J) Representative dendrites spines of PL neurons from sleep-deprived mice after scramble or TRPV4-shRNA lentivirus infusion. Scale bar: 2 μm. (K) Dendritic spine densities of PL neurons in sleep-deprived mice were quantified after different virus infusions ( n ​= ​12–18 dendrites from 3 mice per group). FC: fear conditioned; ASD: acute sleep deprivation; Mean ±SEM. ∗ p ​< ​0.05, ∗∗ p ​< ​0.01, ∗∗∗ p < 0.001. Scale bar: 40 μm.

Article Snippet: Control lentivirus and TRPV4 shRNA lentivirus (approximately 1 μL at a rate of 80 nL/min) were injected bilaterally into the PL (2.34 mm anterior to the bregma, 0.20 mm lateral to the midline, and 1.90 mm ventral to the dura) using a glass pipette connected to a microinjection pump (RWD Life Science, Shenzhen, China).

Techniques: Concentration Assay, shRNA, Expressing, Knockdown, Western Blot, Virus, Immunofluorescence

Journal: Advanced Science

Article Title: LIPUS Promotes Calcium Oscillation and Enhances Calcium Dependent Autophagy of Chondrocytes to Alleviate Osteoarthritis

doi: 10.1002/advs.202413930

Figure Lengend Snippet: TRPV4 mediated Ca 2+ signaling participates the regulation of LIPUS to inflammatory chondrocyte. A) Heatmap showing ion channel gene expression in all cells from the mouse ACLR model of PTOA. B) The express distribution of TRPV4 in UMAP plots. C) Immunohistochemical staining of TRPV4 in the mouse knee joint. Calcium transient relative fluorescence intensity of ROIs with LIPUS stimuli in control group D), in TRPV4‐knockdown group E), and in the TRPV4‐blocked group F). Quantification of the number G) ( n = 8), the magnitude H) ( n = 8) of calcium peaks, and the responsive rate of cells I) ( n = 4). J) Immunofluorescence was used to detect the LC3‐puncta after the chondrocytes was co‐incubated with GSK205. Scale bar, 5 µm. K) Quantification of LC3‐puncta in chondrocytes ( n = 4). WB L) and densitometry analysis of Collagen II M) and MMP‐13 N) protein expression of chondrocyte co‐incubated with GSK205 following LIPUS treatment ( n = 3). Data are presented as means ± SD. Statistical analysis was performed using Student's t test. ****( P < 0.0001), ***( P < 0.001), **( P < 0.01), *( P < 0.05), ns (0.05 < P).

Article Snippet: Lipofection was used to deliver 100 nmol L −1 TRPV4 siRNA into chondrocyte by Lipofectamine 3000 (Invitrogen, USA) following the manufacturer's instructions.

Techniques: Gene Expression, Immunohistochemical staining, Staining, Fluorescence, Control, Knockdown, Immunofluorescence, Incubation, Expressing

Journal: Advanced Science

Article Title: LIPUS Promotes Calcium Oscillation and Enhances Calcium Dependent Autophagy of Chondrocytes to Alleviate Osteoarthritis

doi: 10.1002/advs.202413930

Figure Lengend Snippet: TRPV4 mediated Ca 2+ signaling participates the regulation of LIPUS to inflammatory chondrocyte. A) Heatmap showing ion channel gene expression in all cells from the mouse ACLR model of PTOA. B) The express distribution of TRPV4 in UMAP plots. C) Immunohistochemical staining of TRPV4 in the mouse knee joint. Calcium transient relative fluorescence intensity of ROIs with LIPUS stimuli in control group D), in TRPV4‐knockdown group E), and in the TRPV4‐blocked group F). Quantification of the number G) ( n = 8), the magnitude H) ( n = 8) of calcium peaks, and the responsive rate of cells I) ( n = 4). J) Immunofluorescence was used to detect the LC3‐puncta after the chondrocytes was co‐incubated with GSK205. Scale bar, 5 µm. K) Quantification of LC3‐puncta in chondrocytes ( n = 4). WB L) and densitometry analysis of Collagen II M) and MMP‐13 N) protein expression of chondrocyte co‐incubated with GSK205 following LIPUS treatment ( n = 3). Data are presented as means ± SD. Statistical analysis was performed using Student's t test. ****( P < 0.0001), ***( P < 0.001), **( P < 0.01), *( P < 0.05), ns (0.05 < P).

Article Snippet: To knockdown TRPV4, scramble small‐interfering RNA (siRNA) and TRPV4 siRNA were constructed by RiboBio (Guangzhou, China).

Techniques: Gene Expression, Immunohistochemical staining, Staining, Fluorescence, Control, Knockdown, Immunofluorescence, Incubation, Expressing