vip e el h2155  (Elabscience Biotechnology)

Journal: International Journal of Molecular Medicine

Article Title: Apelin-13 in the paraventricular nucleus (PVN) attenuates myocardial ischemia through V1a receptors in PVN/nucleus tractus solitarii (NTS) and GARγ2 in NTS

doi: 10.3892/ijmm.2025.5652

Figure Lengend Snippet: Effects of apelin-13 microinjection into the PVN on cardiac function in MI model rats. (A) MI models were established one day after implanting an osmotic mini-pump delivering apelin-13 into the PVN. Cardiac function was assessed and tissue samples were collected 28 days after mini-pump implantation. (B) Representative western blots for the V1a receptor and GARγ2 in the PVN and NTS from MI model rats and MI model rats continuously microinjected with apelin-13 into the PVN for 28 days. (C) Quantification of V1a receptor protein levels. (D) Quantification of GARγ2 receptor protein levels. (E) Representative ultrasound images (M-mode) performed on days 28 after MI and sham-operated control. Echocardiographic results for (F) LVEDD, (G) LVESD, (H) LVEF and (I) LVFS (n=6 in the sham-operated control group; n=5 in the MI group, one rat succumbed at 20 days; n=6 in the MI + apelin group). Plasma levels of (J) SST, (K) CCK, (L) VIP, (M) GLP-1, (N) noradrenaline and (O) vasopressin (n=6 in the sham-operated control group; n=5 in the MI group, one rat succumbed at 20 days; n=6 in the MI + apelin group). Normality was tested using the Shapiro-Wilk test. The data in C-D show mean ± SEM; F-O show mean ± SD. Statistical significance was determined by one-way ANOVA, followed by Tukey's multiple-comparisons test. * P<0.05; ** P<0.01; *** P<0.001. Post-hoc power exceeded 80% for all comparisons. PVN, paraventricular nucleus; MI, myocardial infarction; V1a, Vasopressin 1a; GAR, GABA A receptor; NTS, nucleus tractus solitarii; LVEDD, left ventricular end-diastolic diameter; LVESD, lower left ventricular end-systolic diameter; LVEF, left ventricular ejection fraction; LVFS, left ventricular fraction shortening; SST, isomatostatin; CCK, cholecystokinin; VIP, vasoactive intestinal peptide; GLP-1, glucagon-like peptide 1.

Article Snippet: The levels of somatostatin, cholecystokinin, glucagon-like peptide-1 and vasoactive intestinal peptide in the serum were quantified using enzyme-linked immunosorbent assay (ELISA) kits: Somatostatin (cat. no. NBP2-80271; Novus Biologicals; Bio-Techne), cholecystokinin (cat. no. EIAR-CCK-1; RayBiotech, Inc.), glucagon-like peptide-1 (cat. no. E-EL-R3007-96T; Wuhan Elabscience Biotechnology Co., Ltd. Wuhan Elabscience Biotechnology Co., Ltd.), and vasoactive intestinal peptide (cat. no. NBP2-82466; Novus Biologicals; Bio-Techne).

Techniques: Microinjection, Western Blot, Control, Clinical Proteomics

Journal: International Journal of Molecular Medicine

Article Title: Apelin-13 in the paraventricular nucleus (PVN) attenuates myocardial ischemia through V1a receptors in PVN/nucleus tractus solitarii (NTS) and GARγ2 in NTS

doi: 10.3892/ijmm.2025.5652

Figure Lengend Snippet: Mechanisms of the effect of apelin-13 overexpression (AAV2-apelin-13 gene transfer) in the PVN on V1a receptor-mediated improvement in cardiac function in MI model rats. (A) Representative western blot image of V1a receptor expression in the PVN or NTS and GARγ2 expression in the NTS. (B-D) Quantification of V1a receptor expression. (E) Representative western blotting to assess Bcl-2, Bax, TGF-β1 and Smad2 protein levels in the heart and (F-I) quantification of the protein levels. Plasma levels of (J) SST, (K) CCK, (L) VIP and (M) GLP-1 (n=7 in the sham-operated control group; n=5 in the MI group, 2 rats succumbed; n=7 in the MI + apelin-13 group; n=5 in the MI + apelin-13 + SR49059 (PVN) group, 2 rats succumbed; n=6 in the MI + apelin-13 + SR49059 (NTS) group, 1 rat succumbed; n=6 in the MI + apelin-13 + muscimol (NTS) group, 1 rat succumbed). Representative plasma levels of (N) noradrenaline and (O) vasopressin (n=7 in the sham-operated control group; n=6 in the MI group, 1 rat succumbed; n=7 in the MI + apelin-13 group; n=6 in the MI + apelin-13 + SR49059 (PVN) group, 1 rat succumbed; n=6 in the MI + apelin-13 + SR49059 (NTS) group, 1 rat succumbed; n=7 in the MI + apelin-13 + muscimol (NTS) group). Normality was tested using the Shapiro-Wilk test. The data in B-D and F-I show mean ± SEM; data in J-O are shown as mean ± SD and were analyzed via one-way ANOVA, followed by Tukey's multiple-comparisons test. In B-D and F-O, * P<0.05 vs. sham-operated control group; † P<0.05 vs. MI group; ‡ P<0.05 vs. MI + apelin-13 group; § P<0.05 vs. MI + apelin-13 + SR49059 (PVN); # P<0.05 vs. MI + apelin-13 + SR49059 (NTS). Post-hoc power exceeded 80% for all key comparisons. PVN, paraventricular nucleus; V1a, Vasopressin 1a; MI, myocardial infarction; NTS, nucleus tractus solitarii; GAR, GABA A receptor; SST, isomatostatin; CCK, cholecystokinin; VIP, vasoactive intestinal peptide; GLP-1, glucagon-like peptide 1.

Article Snippet: The levels of somatostatin, cholecystokinin, glucagon-like peptide-1 and vasoactive intestinal peptide in the serum were quantified using enzyme-linked immunosorbent assay (ELISA) kits: Somatostatin (cat. no. NBP2-80271; Novus Biologicals; Bio-Techne), cholecystokinin (cat. no. EIAR-CCK-1; RayBiotech, Inc.), glucagon-like peptide-1 (cat. no. E-EL-R3007-96T; Wuhan Elabscience Biotechnology Co., Ltd. Wuhan Elabscience Biotechnology Co., Ltd.), and vasoactive intestinal peptide (cat. no. NBP2-82466; Novus Biologicals; Bio-Techne).

Techniques: Over Expression, Western Blot, Expressing, Clinical Proteomics, Control

Journal: International Journal of Molecular Medicine

Article Title: Apelin-13 in the paraventricular nucleus (PVN) attenuates myocardial ischemia through V1a receptors in PVN/nucleus tractus solitarii (NTS) and GARγ2 in NTS

doi: 10.3892/ijmm.2025.5652

Figure Lengend Snippet: Graphical model summarizing the hypothesized pathway. Overexpression of apelin-13 in the PVN promotes upregulation of V1a receptors in both the PVN and NTS, while downregulating GARγ2 in the NTS. These neural modifications collectively enhance parasympathetic outflow. Furthermore, paraventricular endocrine system secretes circulating neurohumoral factors that systemically mediate cardioprotection. PVN, paraventricular nucleus; V1a, Vasopressin 1a; MI, myocardial infarction; NTS, nucleus tractus solitarii; GAR, GABA A receptor; SST, isomatostatin; CCK, cholecystokinin; GLP-1, glucagon-like peptide 1; VIP, vasoactive intestinal peptide.

Article Snippet: The levels of somatostatin, cholecystokinin, glucagon-like peptide-1 and vasoactive intestinal peptide in the serum were quantified using enzyme-linked immunosorbent assay (ELISA) kits: Somatostatin (cat. no. NBP2-80271; Novus Biologicals; Bio-Techne), cholecystokinin (cat. no. EIAR-CCK-1; RayBiotech, Inc.), glucagon-like peptide-1 (cat. no. E-EL-R3007-96T; Wuhan Elabscience Biotechnology Co., Ltd. Wuhan Elabscience Biotechnology Co., Ltd.), and vasoactive intestinal peptide (cat. no. NBP2-82466; Novus Biologicals; Bio-Techne).

Techniques: Over Expression

Journal: Molecular Pharmacology

Article Title: Extracellular ATP increases agonist potency and reduces latency at class B G protein-coupled receptors

doi: 10.1016/j.molpha.2025.100040

Figure Lengend Snippet: Extracellular ATP enhances cyclic AMP (cAMP) accumulation in response to activation of PAC1R, VPAC1R, and VPAC2R. (A, D, G, and J) COS-7 cells were cotransfected with pGloSensor-22F cAMP biosensor plasmid and a plasmid encoding PAC1R (A), VPAC1R (D), or VPAC2R (G). SaOS-2 cells were transfected with pGloSensor-22F cAMP biosensor plasmid (J). At time 0, cells were stimulated with 0.1 nM PACAP, 0.1 nM VIP, or vehicle (Veh 1 ) in the presence of ATP (1.5 mM) or its vehicle (Veh 2 ). (B, E, H, and K) Cells were stimulated with the indicated concentration of PACAP, VIP, or vehicle (Veh 1 ) in the presence of ATP (1.5 mM) or vehicle (Veh 2 ). Maximum rate of cAMP accumulation was determined as the maximum slope of each cAMP level vs time curve. Data were normalized as a fraction of the greatest value of cyclase activity in each experiment. Values are means ± SEM ( n = 3-5 independent experiments, each performed in triplicate). pEC 50 values for PACAP or VIP in the presence of ATP were significantly greater than the pEC 50 values for PACAP or VIP alone (based on extra sum-of-squares F test, ). (C, F, I, and L) Time-to-maximum-slope was evaluated for the indicated concentration of PACAP or VIP in the presence of ATP (1.5 mM) or vehicle (Veh 2 ). Values are means ± SEM ( n = 3-5 independent experiments, each performed in triplicate). The asterisk (∗) indicates significant difference between agonist + ATP and agonist + Veh 2 ( P < .05, based on 2-way analysis of variance and Bonferroni test). PACAP, pituitary adenylyl cyclase–activating polypeptide; PAC1R, pituitary adenylyl cyclase–activating polypeptide type 1 receptor 1; VIP, vasoactive intestinal peptide; VPAC1R, vasoactive intestinal peptide receptor 1; VPAC2R, vasoactive intestinal peptide receptor 2.

Article Snippet: Salmon calcitonin, pituitary adenylate cyclase–activating peptide (PACAP-38), and vasoactive intestinal peptide (VIP) were purchased from Bachem.

Techniques: Activation Assay, Plasmid Preparation, Transfection, Concentration Assay, Activity Assay

Journal: Molecular Pharmacology

Article Title: Extracellular ATP increases agonist potency and reduces latency at class B G protein-coupled receptors

doi: 10.1016/j.molpha.2025.100040

Figure Lengend Snippet: Extracellular ATP enhances PACAP-induced recruitment of β -arrestin-1 to PAC1R. (A) HEK293H cells were cotransfected with plasmids encoding N-terminal luciferase fragment- β -arrestin-1 and PAC1R-C-terminal luciferase fragment. At time 0, cells were stimulated with the indicated concentration of PACAP (10 nM) or vehicle (Veh 1 ) in the presence of ATP (1.5 mM) or its vehicle (Veh 2 ). Luminescence intensity, which corresponds to β -arrestin-1 recruitment to PAC1R, was then measured from live cells every 2 minutes. Panel illustrates the results of individual experiments, representative of 3 independent experiments each performed in triplicate. (B) Cells were stimulated with indicated concentrations of PACAP or its vehicle (Veh 1 ). The maximum rate of β -arrestin-1 recruitment was determined from the β -arrestin-1 recruitment vs time curves as the greatest slope. Data were normalized as a fraction of the greatest rate of recruitment in each experiment. Values are means ± SEM ( n = 3 independent experiments, each performed in triplicate). pEC 50 for PACAP in the presence of ATP was significantly greater than pEC 50 for PACAP alone (based on extra sum-of-squares F test, ). PACAP, pituitary adenylyl cyclase–activating polypeptide; PAC1R, pituitary adenylyl cyclase–activating polypeptide type 1 receptor 1.

Article Snippet: Salmon calcitonin, pituitary adenylate cyclase–activating peptide (PACAP-38), and vasoactive intestinal peptide (VIP) were purchased from Bachem.

Techniques: Luciferase, Concentration Assay

Journal: Experimental Physiology

Article Title: Intact microdissection of stellate ganglia in a Parkinson's disease model reveals aggregation of mutant human α‐synuclein in their cell bodies

doi: 10.1113/EP092261

Figure Lengend Snippet: Sympathetic and parasympathetic marker expression in stellate ganglion. Sympathetic marker TH and parasympathetic marker VIP were co‐stained in stellate ganglion. Nuclei were stained with DAPI. (a) Stellate ganglion. TH was detected in neuronal cell bodies. (b) Higher magnification of a neuronal cell body (white arrow). (c1) Positive control, cerebral cortex. (c2) Positive control, brainstem. (d1) Higher magnification of c1, TH‐positive cell body (yellow arrow). (d2) Higher magnification of c2, VIP‐positive cells (magenta arrow). (e) Negative control in stellate ganglion. (f) Higher magnification of (e). Scale bar 100 µm in (a, c1, c2, e); scale bar 20 µm in (b, d1, d2, f). DAPI, 4′,6′‐diamidino‐2‐phenylindole; TH, tyrosine hydroxylase; VIP, vasoactive intestinal peptide.

Article Snippet: All the antibodies were diluted in 1% normal bovine serum in PBS, and the dilution rates were the following: α‐synuclein human monoclonal antibody‐15G7 (1:125 dilution, overnight at −4°C, cat. no. ALX‐804‐258, Enzo Life Science, Farmingdale, NY, USA), tyrosine hydroxylase (TH) polyclonal antibody (1:250, cat. no. PA5‐18372, Thermo Fisher Scientific, Waltham, MA, USA), vasoactive intestinal peptide (VIP) polyclonal antibody (1:250, cat. no. PA1‐85958, Thermo Fisher Scientific), neuronal nitric oxide synthase (nNOS) polyclonal antibody (1:250, overnight, PA1‐033, Thermo Fisher Scientific), anti‐α‐synuclein aggregate [MJFR‐14‐6‐4‐2] (1:5000, 2 h, cat. no. ab209538, Abcam, Cambridge, UK), and anti‐Thy‐1 monoclonal (1:500, overnight, cat. no. ab307736, Abcam).

Techniques: Marker, Expressing, Staining, Positive Control, Negative Control