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8 protocols using sr49059

1

Competitive V1a Receptor Antagonist in Cerebral Ischemia

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SR49059 (Sigma-Aldrich, St Louis, MO, USA), the most potent nonpeptide competitive V1aR antagonist,31 (link) was initially dissolved in dimethyl sulphoxide at a concentration of 30 mM, and then diluted with 0.9% NaCl to 0.3 mM as a finial concentration. The drug application timeline was shown in Supplementary Figure S1. Since the 15th day after 2VO surgery, the 2VO + HTS group was given 5.3% NaCl (refs 32 (link), 33 (link)) as a single intraperitoneal injection of 2% bodyweight, 1 h before the Morris water maze (MWM) or LTP every day.32 (link), 34 (link) The 2VO + HTS + SR49059 group was given 0.3 mg kg−1 (1.67 ml kg−1, 0.3 mM) SR49059 (refs 35 , 36 (link)) intraventrally through the tail vein at 30 min following injection of 5.3% NaCl as described above, and the sham and 2VO groups received 0.9% NaCl as control. Water bottles were removed at the moment of first injection for all groups until 10 min before MWM and LTP.
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2

Vasopressin and SR49059 Protocol

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Vasopressin and SR49059 were obtained from Sigma-Aldrich and dissolved in ultrapure distilled water on the day of the study.
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3

Avian Chick Photoperiod Experiment

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One-day old male chicks (Cobb 500) were obtained from a commercial hatchery and raised in an environmentally controlled room and fed a standard starter diet ad libitum. Birds were maintained on continuous light (L) with no dark (D) periods (LD 24:0) for 3 days, and then moved to a long-day photoperiod of LD 16:8. The selected mammalian V1bR antagonists (SSR- 149415, Manning compound, L-368899) and V1aR antagonist (SR-49059) were purchased from Sigma-Aldrich (St. Louis, MO, USA), and AVT and CRH were purchased from Bachem Americas Inc. (Torrance, CA, USA).
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4

Arginine Vasopressin Antagonist Study

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Arginine vasopressin (AVP), SR49059, neutral red, and Hexamethonium were purchased from Sigma, USA, and chloral hydrate and NaCl were purchased from Tiancheng, Chemical Ltd., China. All chemicals were in technical grade of more than 95% purity. All drugs were dissolved in normal saline (NS) and prepared immediately before use.
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5

Peptide and Pharmacological Compounds Procurement

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MTII and alpha-MSH were purchased from Bachem (Torrence, CA, USA). 5′-AMP, 8-cyclopentyl-1,3-dimethylxanthine (CPT), isoproterenol (Iso), and SR49059 (SR) were purchased from Sigma (St. Louis, MO, USA).
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6

Comparative Evaluation of Vasopressor Agents

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SR-49059 (Relcovaptan) is a selective nonpeptidic V1a receptor antagonist. It was obtained from Sanofi-Aventis (Sanofi-Aventis, Toulouse, France). SR-49059 was dissolved in DMSO (48 mg/mL) then extemporaneously emulsified in Cremophor® RH40 (Sigma-Aldrich, Saint-Quentin-Fallavier, France) and diluted in 9% sodium chloride solution before injection. Treatment consisted of three intraperitoneal injections of SR-49059 solution performed, 1 h before anesthesia (4.5 mg/kg), 8 h after the surgery (6 mg/kg) and 1 h before sacrifice (4.5 mg/kg).
Terlipressin is a well-documented vasopressin receptor agonist inducing intense and prolonged vasoconstriction. Commercial terlipressin, Glypressine®, was obtained from Ferring GmbH (Kiel, Germany). Its purity has been previously evaluated by mass spectrometry [30 (link)]. Terlipressin solution (20 µg/mL) was prepared by solubilization in 9% sodium chloride. Treatment consisted of three intraperitoneal injections of terlipressin solution performed 1, 12 and 23 h after surgery and corresponding to a total dose of 80 µg/kg/24 h. To reduce the risk of bleeding due to the increase of blood pressure, terlipressin was not administered either preoperatively or immediately after surgery.
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7

Murine Allergic Inflammation Modulation

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Six to eight-week-old female mice (BALB/c, 20-24 g) obtained from Shanghai Experimental Animal Center (Shanghai, China) were given standard diet and water. All the experimental protocols were approved by the Animal Ethics Committee of Shanghai Jiao Tong University. Rhodamine (X-rhodamine-5-(and-6)isothiocyanate), ovalbumin (OVA, Grade V), SR-49059 (V1A antagonist), SR-121463A (V2 antagonist), KT-5720 (PKA inhibitor), Go-7874 (PKC inhibitor), hexadecyltrimethylammonium bromide (HTAB), and 3, 3', 5, 5'-tetramethylbenzidine (TMB) were collected from Sigma-Aldrich (St Louis, MO). Southern Biotech (Birmingham,USA) provided HRP labelled anti-mouse IgE secondary. ELISA kits of IL-6, IL-10, and IFN-γ were supplied from Biolegend (San Diego, CA)
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8

Pharmacological Agents for Research

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ELA-21 and F13A were obtained from Phoenix Pharmaceuticals (Belmont, CA). Hexamethonium, SR49059, losartan, and LY294002 were purchased from Sigma Chemical (St. Louis, MO). MK-2206 was obtained from Selleck Chemicals (Houston, TX).
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