rns® system  (NeuroPace)

Journal: Cancer Science

Article Title: The Serotonergic and Cholinergic Signaling Pathways Drive Gastric Precancerous Lesions and Its Potential Mechanism

doi: 10.1111/cas.70563

Figure Lengend Snippet: 5‐HTR1D and TPRV4 were involved in 5‐HT‐induced currents in gastric cells. Representative vehicle control recordings showing that application of the vehicle (≤ 5% DMSO in extracellular solution) alone did not induce detectable membrane currents in GES‐1 (a), AGS (b), or MKN45 (c) cells. (d) The change rate of currents induced by vehicle in GES‐1, AGS, and MKN45 cells. (e–g) Representative whole‐cell currents recorded in GES‐1 (e), AGS (f), and MKN45 (g) cells in response to 5‐HT (30 μM) and subsequent inhibition by the selective 5‐HTR1D antagonist LY310762 (10 μM). (h) The change rate of currents induced by LY‐310762 in GES‐1, AGS, and MKN45 cells. (i–k) Representative whole‐cell current recordings in GES‐1 (i), AGS (j), and MKN45 (k) cells showing the response to 5‐HT (30 μM) and its suppression by the selective TRPV4 antagonist RN‐1734 (10 μM). (l) The change rate of current induced by RN‐1734 in GES‐1, AGS, and MKN45 cells. (m,q) qRT‐PCR analysis of TRPV4 mRNA expression following transfection with three independent TRPV4 siRNA sequences in AGS and MKN45 cells, respectively; siRNA‐2 exhibited the highest knockdown efficiency and was used for subsequent electrophysiology experiments. (n–p) Representative whole‐cell currents evoked by 5‐HT in AGS cells transfected with negative control siRNA or TRPV4 siRNA‐2, and corresponding quantification of peak current amplitudes (p). (r–t) Representative 5‐HT‐triggered current traces from MKN45 cells transfected with negative control siRNA or TRPV4 siRNA‐2, along with statistical analysis of maximum current magnitudes (t). Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 versus GES‐1 or scrambled siRNA control.

Article Snippet: The 5‐HTR1D antagonist group was administered LY310762 (1 mg/kg/day in 0.4 mL vehicle; Cat. No. GC17138, Glpbio, USA) [ ], and the TRPV4 antagonist group received RN‐1734 (10 μmol/L in 0.4 mL vehicle; Cat. No. HY‐19975, MedChemExpress, USA).

Techniques: Control, Membrane, Inhibition, Quantitative RT-PCR, Analysis, Expressing, Transfection, Knockdown, Negative Control

Journal: Cancer Science

Article Title: The Serotonergic and Cholinergic Signaling Pathways Drive Gastric Precancerous Lesions and Its Potential Mechanism

doi: 10.1111/cas.70563

Figure Lengend Snippet: 5‐HTR1D and TRPV4 are involved in intestinal metaplasia and dysplasia of stomach with concomitant reduction of AKT/mTOR activation. (a) Schematic diagram of the GPL modeling protocol and experimental timeline. (b) Administration of 5‐HTR1D antagonist or TRPV4 antagonist significantly attenuated the GPL‐induced decrease in body weight ( n = 12). (c) Representative gastric mucosal morphology of mice in 4 groups. (d) Representative AB‐PAS staining of gastric mucosa of mice in 4 groups. (e) 5‐HTR1D mRNA expression in gastric tissues from control and GPL mice, determined by qRT‐PCR ( n = 6). (f) Representative Western blot bands showing protein levels of AKT, p‐AKT, mTOR, and p‐mTOR in gastric mucosal tissues of 4 groups. (g–j) Quantitative analysis of AKT (g), p‐AKT (h), mTOR (i), and p‐mTOR (j) expression levels ( n = 6). * p < 0.05, ** p < 0.01, *** p < 0.001 versus Control group; # p < 0.05, ## p < 0.01 versus GPL group.

Article Snippet: The 5‐HTR1D antagonist group was administered LY310762 (1 mg/kg/day in 0.4 mL vehicle; Cat. No. GC17138, Glpbio, USA) [ ], and the TRPV4 antagonist group received RN‐1734 (10 μmol/L in 0.4 mL vehicle; Cat. No. HY‐19975, MedChemExpress, USA).

Techniques: Activation Assay, Morphology, Staining, Expressing, Control, Quantitative RT-PCR, Western Blot, Analysis

Journal: Cancer Science

Article Title: The Serotonergic and Cholinergic Signaling Pathways Drive Gastric Precancerous Lesions and Its Potential Mechanism

doi: 10.1111/cas.70563

Figure Lengend Snippet: Expression of 5‐HT and ACh and M3R/5‐HT co‐localization in gastric antral tissues of GPL model mice. (a) 5‐HT levels were measured by ELISA. (b) Representative images showing M3R/5‐HT co‐localization in gastric antral mucosa, as detected by immunofluorescence. (c) The increased percentage of M3R/5‐HT co‐localization neurons in GPL mice. (d) ACh expression was determined using ELISA. (e) The schematic diagram of FG retrograding test. (f) The FG labeling ChAT‐positive neurons in the DMV 7 days after FG injection ( n = 6). (g) Representative images showing c‐Fos expression of ChAT‐positive neurons in control, GPL, LY310762 and RN1734 groups. (h) The percentage of c‐Fos + DMV cholinergic neurons in all groups. Data are presented as mean ± SD ( n = 6). Scale bar: 100 μm. * p < 0.05, ** p < 0.01, *** p < 0.001 versus Control; # p < 0.05, ## p < 0.01 versus GPL group.

Article Snippet: The 5‐HTR1D antagonist group was administered LY310762 (1 mg/kg/day in 0.4 mL vehicle; Cat. No. GC17138, Glpbio, USA) [ ], and the TRPV4 antagonist group received RN‐1734 (10 μmol/L in 0.4 mL vehicle; Cat. No. HY‐19975, MedChemExpress, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Labeling, Injection, Control

Journal: Cancer Science

Article Title: The Serotonergic and Cholinergic Signaling Pathways Drive Gastric Precancerous Lesions and Its Potential Mechanism

doi: 10.1111/cas.70563

Figure Lengend Snippet: The graphical abstract of the proposed mechanism through which the central neural system may modulate the GPL process. Vagal cholinergic neurons may promote gastric 5‐HT secretion, which may contribute to gastric precancerous lesions through 5‐HTR1D and TRPV4, highlighting these as candidate targets for early intervention.

Article Snippet: The 5‐HTR1D antagonist group was administered LY310762 (1 mg/kg/day in 0.4 mL vehicle; Cat. No. GC17138, Glpbio, USA) [ ], and the TRPV4 antagonist group received RN‐1734 (10 μmol/L in 0.4 mL vehicle; Cat. No. HY‐19975, MedChemExpress, USA).

Techniques: