Post-weaned male rats (50–55 g) were anaesthetized with a ketamine (75 mg kg−1 i.p.)/xylazine (5 mg kg−1 i.p.) mixture. The depth of anesthesia was checked at regular intervals (20–30 min) by assessing the withdrawal reflex response to noxious pinching of the tail or hind paw. Animals were placed in a stereotaxic frame (tooth bar −3.5 mm below the inter-aural line; David Kopf, Tujunga, USA) and two microinjections (different rostrocaudal levels separated by 300 μm) per side of either PRSx8-AlstR-GFP-LVV or PRSx8-GFP-LVV (50 nl each, over 5 min) were delivered into RVLM (Picospritzer II; Parker Instruments, Cleveland, USA). The injection pipette was angled at 25° and injections were made −3.7 mm ventral from calamus scriptorius and ± 1.7 mm lateral from the midline. Post-surgery, rats were treated with one prophylactic dose of analgesic and antipyretic flunixin meglumine (1 mg kg−1; Schering-Plough, Rio de Janeiro, Brazil) and 0.1 ml of veterinary antibiotic (1.2 million i.u.; Fort Dodge, Campinas, Brazil) via intramuscular injections.
Flunixin meglumine
Manufactured by Merck & Co
Sourced in Brazil
Flunixin meglumine is a pharmaceutical compound used as a laboratory reagent. It is a nonsteroidal anti-inflammatory drug (NSAID) that possesses analgesic, anti-inflammatory, and antipyretic properties. Flunixin meglumine is commonly used in research applications, but its specific core function should be determined based on the intended use of the product.
Automatically generated - may contain errors
Lab products found in correlation
Urethane, by Merck Group (4 mentions)
Xylazine, by Syntec (1 mentions)
Lidocaine, by Dentsply (1 mentions)
Ketamine, by Syntec (1 mentions)
Isoflurane, by Merck & Co (1 mentions)
Cp376395, by Bio-Techne (1 mentions)
Isoforine, by Cristália (1 mentions)
Losartan potassium, by Bio-Techne (1 mentions)
Pd123319 ditrifluoroacetate, by Bio-Techne (1 mentions)
Tribromoethanol, by Merck Group (1 mentions)
Rx821002, by Bio-Techne (1 mentions)
Wb4101, by Bio-Techne (1 mentions)
Propranolol, by Merck Group (1 mentions)
2 2 2 tribromoethanol, by Merck Group (1 mentions)
12 protocols using flunixin meglumine
1
Stereotaxic Targeting of RVLM in Rats
2
Stereotaxic Surgical Implantation of Cannulae in Rat Prefrontal Cortex
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Animals were deeply anesthetized using 1.0 ml/kg of a solution of xylazine (10 mg/ml; Syntec, Brazil) and ketamine (100 mg/ml; Syntec, Brazil), placed in a stereotaxic frame, then local anesthesia with lidocaine 3.0% and epinephrine 1:50,000 (Dentsply, Brazil) was applied. Two stainless steel guide cannulae (22 G, 11 mm) were implanted aimed at the IL cortex following coordinates (AP: + 3.2 mm from Bregma, ML: + 0.6 mm, DV: − 2.1 mm) from the rat brain atlas by Paxinos and Watson26 , and fixed to the skull using acrylic resin and two stainless steel screws. An obturator (30G) was inserted inside each guide cannula to reduce possible occlusion. For post-surgery analgesia, anti-inflammatory, and antipyretic actions, animals received an intramuscular injection of flunixin meglumine (2.5 mg/kg; Schering-Plough, Brazil). Animals were given at least ten days of recovery before behavioral testing.
3
Stereotaxic Electrode Implantation in Rats
4
Antagonist Receptor Binding and Anesthesia
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The selective CRF1 receptor antagonist CP376395 (Tocris, Westwoods Business Park, Ellisville, Missouri, USA), corticotropin-releasing factor (CRF) (Sigma-Aldrich, St. Louis, Missouri, USA), and urethane (Sigma-Aldrich) were dissolved in saline (0.9% NaCl). The veterinary pentabiotic (Fort Dodge, Campinas, SP, Brazil), isoflurane (Cristrália, Itapira, São Paulo, Brazil), and the nonsteroidal anti-inflammatory flunixin meglumine (Banamine, Schering-Plough, Cotia, SP, Brazil) were used as supplied by the manufacturers.
5
Pharmacological Modulation of Angiotensin Receptors
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Losartan potassium (selective AT1 receptor antagonist) (Tocris, Westwoods Business Park, Ellisville, Missouri, USA), A-779 (selective MAS receptor antagonist) (Tocris), PD123319 ditrifluoroacetate (selective AT2 receptor antagonist) (Tocris) and urethane (Sigma-Aldrich, St Louis, Missouri, USA) were dissolved in saline (NaCl 0.9%). The poly-antibiotic formulation (Pentabiotico; Fontoura-Wyeth, Campinas, São Paulo, Brazil), isoflurane (Isoforine; Cristália, Itapira, São Paulo, Brazil), lidocaine (Harvey Química Farmacêutica Ind. e Comércio Ltd.a., Catanduva, São Paulo, Brazil), and flunixin meglumine (Banamine, Schering Plough, Cotia, São Paulo, Brazil) were used as provided.
6
Adrenoceptor Antagonist Cocktail Protocol
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WB4101 (selective α1-adrenoceptor antagonist; Tocris, Westwoods Business Park, Ellisville, MO, USA; cat. # 0946), RX821002 (selective α2-adrenoceptor antagonist; Tocris, cat. # 1324), propranolol (β-adrenoceptor antagonist; Sigma–Aldrich, St. Louis, Missouri, USA; cat. # P0884), tribromoethanol (Sigma–Aldrich, cat. # T48402), and urethane (Sigma–Aldrich; cat. # U2500) were dissolved in saline (NaCl 0.9%). Flunixin meglumine (Banamine, Schering Plough, Cotia, São Paulo, Brazil) and the polyantibiotic preparation of streptomycins and penicillins (Pentabiotico, Fort Dodge, Campinas, São Paulo, Brazil) were used as provided.
7
Targeted Silencing of Locus Coeruleus Neurons
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Male rats (225–250 g) were anaesthetised with a ketamine (75 mg kg−1 i.p.)/xylazine (5 mg kg−1 i.p.) mixture. The depth of anaesthesia was checked at regular intervals (15–20 min) by assessing the withdrawal reflex response to noxious pinching of the tail or hind paw. Animals were placed in a stereotaxic frame (David Kopf, Tujunga, USA) and four microinjections (each 250 μm apart in the dorsoventral axis) per side of PRSx8-AlstR-GFP-LVV (250 nl each, over 2 min) were delivered into LC bilaterally (Picospritzer II; Parker Instruments, Cleveland, USA). The microinjections were made 2 mm caudal to bregma, ±1.2 mm lateral from the midline and 5.5–6.0 mm below the brain surface with a 10° rostral angulation. A stainless-steel guide cannula (13 mm long, 0.6 mm o.d., 0.4 mm i.d.) was implanted into the lateral cerebral ventricle (−0.6 mm to Bregma, 1.5 mm lateral to the midline and −3.6 mm ventral to dura mater). The guide cannula was fixed to the cranium using dental acrylic resin. Post-surgery, rats were treated with one prophylactic dose of analgesic and antipyretic flunixinmeglumine (1 mg/kg; Schering-Plough, Rio de Janeiro, Brazil) and 0.1 ml of veterinary antibiotic (1.2 million i.u.; Fort Dodge, Campinas, Brazil) via intramuscular injections.
8
Generating Parabiotic Mouse Pairs
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To generate parabiotic pairs, we followed our previously described procedure (Casanova-Acebes et al., 2013 (link)). Anesthetized mice were shaved at the corresponding lateral aspects and matching skin incisions were made from the olecranon to the knee joint of each mouse, and the subcutaneous fascia was bluntly dissected to create ∼0.5 cm of free skin. The olecranon and knee joints were attached by a single 5-0 polypropylene suture and tie, and the dorsal and ventral skins were approximated by continuous suture. A single dose of flunixin-meglumine (Schering-Plough) was injected subcutaneously in each partner at the end of the surgical procedure (1 mg/kg). 1 mo after surgery, samples were obtained from each of the partners for analyses. To ensure that partner-derived neutrophils in tissues derived from the circulation rather than from partner progenitor cells that could have also crossed to the host mouse, we generated parabionts that were separated after 1 mo. In the separated mice we confirmed a rapid decline of partner-derived neutrophils which was reduced from 15.5% to 4% in blood and tissues after 4 d, which indicated that the majority of neutrophils found in tissues of the host mice derived from the circulation.
9
Skin Flap Graft Procedure in Mice
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Anaesthetized mice were shaved at the corresponding lateral aspects and matching skin incisions were made from the olecranon to the knee joint of each mouse, and the subcutaneous fascia was bluntly dissected to create ~0.5 cm of free skin. The olecranon and knee joints were attached by a single 5–0 polypropylene suture and tie, and the dorsal and ventral skins were approximated by continuous suture. A single dose of flunixin meglumine (Schering-Plough, 1 mg kg−1) was injected subcutaneously into each partner at the end of the surgical procedure. One month after surgery, blood samples were obtained from each of the partners for analysis of haematopoietic progenitors.
10
Mouse Parabiosis Surgical Procedure
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To generate parabiotic pairs, we followed our previously described procedures (A-Gonzalez et al., 2017) . Anesthetized mice were shaved at the corresponding lateral aspects and matching skin incisions were made from the olecranon to the knee joint of each mouse, and the subcutaneous fascia was bluntly dissected to create about 0.5 cm of free skin. The olecranon and knee joints were attached by a single 5-0 polypropylene suture and tie, and the dorsal and ventral skins were approximated by continuous suture. A single dose of flunixin meglumine (Schering-Plough, Segre, France) was injected subcutaneously in each partner at the end of the surgical procedure (1 mg/kg). One month after surgery blood samples were obtained from each of the partners to assess leukocyte chimerism.
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