The mice were anesthetized by inhalation of Isoflurane (Isoflurane, Abbott, Maidenhead, UK; dosages detailed in the MCAO procedure description). The spleen was removed via a dorsolateral approach, the fur around the 13th rib was shaved, the area was disinfected with 70% ethanol, and an incision was made below the 13th rib to access the spleen. It was separated from its blood vessels using electrocautery, and the wound was closed with metal clips. The animals were monitored for one day in a heated cage with analgesia using 0.1 mL of buprenorphine (Buprenovet, Bayer, Leverkusen, Germany). Full recovery from the operation was deemed after a week (22 (link),23 (link)).
Buprenorphine
Manufactured by Bayer
Sourced in Germany
Buprenorphine is a laboratory-grade product manufactured by Bayer for use in scientific research and analysis. It is a synthetic opioid compound that can be used as a reference standard or analytical tool in various chemical and pharmaceutical applications. The core function of Buprenorphine is to serve as a reliable and consistent material for research purposes, without any interpretation or extrapolation on its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Baytril, by Bayer (3 mentions)
Buprenovet, by Bayer (2 mentions)
Propofol, by Fresenius (2 mentions)
Ketamine, by Zoetis (2 mentions)
Isoflurane, by Abbott (1 mentions)
Cuy21 sc square wave electroporator, by Nepa Gene (1 mentions)
Fast green, by Merck Group (1 mentions)
Picospritzer 3, by Parker Hannifin (1 mentions)
Rimadyl, by Pfizer (1 mentions)
Carprofen, by CP-Pharma (1 mentions)
Forene, by Abbott (1 mentions)
Enrofloxacin, by Bayer (1 mentions)
Naloxon, by B. Braun (1 mentions)
Perma hand silk suture, by Johnson & Johnson (1 mentions)
Microprocessor controlled minipump, by World Precision Instruments (1 mentions)
Domitor, by Orion Pharma (1 mentions)
Antisedan, by Orion Pharma (1 mentions)
Xylocain, by AstraZeneca (1 mentions)
10 protocols using buprenorphine
1
Spleen Removal in Murine Ischemic Stroke
2
In Utero Electroporation of Mouse Embryos
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In utero electroporation was performed as described before.14 (link) Time pregnant (E14) CD1 wild-type mice were deeply anesthetized with isoflurane. Ketoprofen (5 mg/kg - Mibe) and Buprenorphine (0.05 mg/kg - Bayer) were used as analgesia. Uterine horns were exposed to inject each embryo once into the lateral ventricle with 1–2 μL DNA with a concentration of 1.5 μg/μL and fast green (1 mg/mL, Sigma, USA) using a pulled and beveled glass capillary (Drummond Scientific, USA) and a microinjector (Picospritzer III, Parker Hannifin Corporation, USA). Five electric pulses with 50 ms duration were applied at 950 ms intervals with a triple-electrode using a CUY21SC Square Wave Electroporator charged to 30 V (Nepa Gene, Japan). Electrode forceps were positioned to IUE cortical progenitor cells in the somatosensory and motor cortex.58 (link)
3
Post-Surgical Care and Recovery Monitoring
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After surgery and the two following days the rats received 2–4 ml saline, 0.03 mg/kg buprenorphine (Temgesic, Pharmaceuticals limited, UK), 5 mg/kg carprofen (Rimadyl, Pfizer), 10 mg/kg enrofloxacin (Baytril, Bayer) all given s.c. and 0.4 mg/kg buprenorphine mixed in Nutella (Ferrero, Germany). In order to obtain proper and valid ICP data it is of great importance that the animals recover well from surgery and remain healthy, therefore body weight and water intake was monitored daily for a week post-surgery. Thereafter weight was monitored once a week. The following days after surgery, the rats lost some weight (< 15%) and from day 7 weight and water intake increased (Additional file 1 : Figure S1). It was observed that their general well-being and activity level also increased.
4
Postoperative Care and Monitoring in Animal Model
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All of the animals received buprenorphine (0.3 mg/mL bw; Buprenovet®, Bayer AG, Leverkusen, Germany) as the analgesic medication perioperatively, and for the following 4 days every 12 h. After each procedure, postoperative stress was recorded using a score sheet. These included daily monitoring of body weight, general condition, spontaneous behavior in the cage, and clinical examination (temperature, respiration, pulse, and warmth of extremities). These parameters were checked daily until the animals regained their preoperative bodyweight and a good clinical condition. Furthermore, the animals were explored for clinical signs of infection of the operated limb (swelling, reddening, impairment of wound healing, and loss of passive motion in the left hind leg). Eight weeks after the second surgery, the animals were sacrificed.
5
Transverse Aortic Constriction Mouse Model
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LVPO was induced by TAC in 9‐week‐old mice.
22 (link) Mice were subcutaneously pretreated with 1 mg/kg buprenorphine (Bayer, Leverkusen, Germany) and 5 mg/kg carprofen (Cp‐Pharma, Burgdorf, Germany). Anesthesia was induced with 5% isoflurane, sustained with 2% isoflurane, and monitored by toe pinching. Eye care solution was applied to prevent corneal injury and mice were placed in the supine position on a heating pad (37 °C). Mice were orally intubated and mechanically ventilated using a rodent ventilator (Hugo Sachs Elektronik, March‐Hugstetten, Germany). Following horizontal skin incision at the level of the suprasternal notch, the transverse aortic arch was visualized through a median sternotomy. Next, the transverse aortic arch was ligated using a 6–0 silk suture that was tightened around a blunt 25‐G needle, which was placed between the innominate artery and the left common carotid artery. The needle was quickly removed, leaving a discrete region of stenosis. The chest and overlying skin were closed. Animals received metamizole (1A Pharma, Holzkirchen, Germany) in the drinking water (200 mg/kg) for 4 days total, starting the day before surgery. The sham procedure was performed identically, except no suture was placed around the aortic arch.
22 (link) Mice were subcutaneously pretreated with 1 mg/kg buprenorphine (Bayer, Leverkusen, Germany) and 5 mg/kg carprofen (Cp‐Pharma, Burgdorf, Germany). Anesthesia was induced with 5% isoflurane, sustained with 2% isoflurane, and monitored by toe pinching. Eye care solution was applied to prevent corneal injury and mice were placed in the supine position on a heating pad (37 °C). Mice were orally intubated and mechanically ventilated using a rodent ventilator (Hugo Sachs Elektronik, March‐Hugstetten, Germany). Following horizontal skin incision at the level of the suprasternal notch, the transverse aortic arch was visualized through a median sternotomy. Next, the transverse aortic arch was ligated using a 6–0 silk suture that was tightened around a blunt 25‐G needle, which was placed between the innominate artery and the left common carotid artery. The needle was quickly removed, leaving a discrete region of stenosis. The chest and overlying skin were closed. Animals received metamizole (1A Pharma, Holzkirchen, Germany) in the drinking water (200 mg/kg) for 4 days total, starting the day before surgery. The sham procedure was performed identically, except no suture was placed around the aortic arch.
6
Bilateral Drug Infusion into the Lateral Septum
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Guide cannulas (23 G, 8 mm length; Injecta GmbH) for bilateral drug infusion into the LS were implanted stereotaxically with the tip resting 2 mm above the LS target region (from bregma: anterior–posterior: −0.3 mm, mediolateral: ±0.5 mm, dorsoventral: −1.6 mm [23 ]), under isoflurane anesthesia (Forene, Abbott GmbH) [10 (link), 24 ]. All mice received a subcutaneous injection of antibiotics (10 mg/kg Baytril, Bayer GmbH) and analgesics (0.05 mg/kg Buprenorphine, Bayer) to avoid post-surgical infections and pain. In order to reduce post-surgical stress, all mice were handled once a day for at least 5 days before experiments wherein dummy cannulas closing the guide cannulas were disinfected.
7
Stereotaxic Surgery for Intra-CeA Cannulation
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Stereotaxic surgery was performed under semi-sterile conditions as previously described (De Jong et al., 2014 (link); Oliveira et al., 2021 (link)). Briefly, rats were anesthetized with isoflurane, injected i.p. with the analgesic Buprenovet (0.05 mg/kg Buprenorphine, Bayer, Germany) and the antibiotic Baytril (10 mg/kg Enrofloxacin, Baytril, Bayer, Germany), and mounted in the stereotaxic frame. Intra-CeA (AP: -2.5; ML: ±4.0; DV: +6.0) guide cannulas (25 G, 12 mm; Injecta GmbH, Germany) were implanted bilaterally, 2 mm above the target region to avoid lesioning. Local cannulas were fixed to the skull with two jeweler’s screws and dental cement (Kallocryl, Speiko-Dr. Speier GmbH, Muenster, Germany) and closed by a stainless steel stylet (27 G). Cannulated females were left single-housed and undisturbed for 5 days to recover, prior to the start of the aggression training protocol.
8
Viral Injections in Mouse Hippocampus
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Viral injections were performed under aseptic conditions in Prox1::cre or C57BL/6 mice up to 10 months old. The mice were anesthetized with a mixture of Fentanyl (Rotexmedica, Germany), Midazolam (Rotexmedica, Germany), and Domitor (Orion Pharma, Finland) via intraperitoneal injection ( ). Analgesia ( of buprenorphine; Buprenovet, Bayer, Germany) was administered intraperitoneally prior to the injection, and Xylocain (AstraZeneca, Germany) was used for local anesthesia. Stereotaxic injections were performed using an injection frame (WPI Benchmark/Kopf) and a microprocessor-controlled minipump (World Precision Instruments, Sarasota, Florida), 1000 nl of the viral solution was injected bilaterally into the hippocampus (rAAV-DIO-eGFP; rAAV-syn-GFP; coordinates: rostrocaudal: from the Bregma; mediolateral: from the midline; dorsoventral: ). After the injection, the scalp was sutured with PERMA-HAND Silk Suture (Ethicon), and an antibacterial ointment (Refobacin®, Almirall, Germany) was applied, followed by the intraperitoneal injection of a mixture of Naloxon (B. Braun, Germany), Flumazenil (B. Braun, Germany), and Antisedan (Orion Pharma, Finland) ( ). To prevent the wound pain, analgesia was administered on the 3 following days.
9
Electrophysiological Studies in Anesthetized Pigs
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Electrophysiological studies were performed in 17 anesthetized pigs of both sexes (<6 months of age; body weight 30–45 kg). After sedation with azaperon (5 mg/kg IM; Elanco, Bad Homburg, Germany), midazolam (1 mg/kg IM; Hameln Pharma Plus GmbH, Hameln, Germany), and ketamine (10 mg/kg IM; Zoetis Deutschland GmbH, Berlin, Germany), animals were anesthetized with propofol (1.5 mg/kg IV bolus followed by 4–8 mg/kg/h IV; Fresenius Kabi, Bad Homburg, Germany). For analgesia, buprenorphine (0.02 mg/kg IV; Bayer Vital GmbH Tiergesundheit, Leverkusen, Germany) was administered. Mechanical ventilation was performed using the Draeger Primus system (Draeger, Luebeck, Germany). Before surgical jugular vein preparation, a single dose of cefuroxime was administered (750 mg IV; Ratiopharm GmbH, Ulm, Germany). No volatile anesthetics were used to avoid pharmacologic interaction with cardiac K2P channels. Pigs were kept under specific pathogen‐free conditions at a room temperature of 20°C±2°C with a maximum housing density according to directive 2010/63/EU. Room lighting had a light/dark cycle of 12/12 h. Water was offered ad libitum and pigs were fed twice a day with balanced complete feed (SAF 130M, ZG Raiffeissen, Karlsruhe, Germany). Environmental enrichment was provided with biting woods, chains, and feeding balls.
10
Anesthesia protocol for porcine studies
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Upon sedation with midazolam (1 mg/kg, intramuscular [i.m.]; Hameln Pharma Plus GmbH, Berlin, Germany), azaperon (5 mg/kg, i.m.; Elanco, Bad Homburg, Germany), and ketamine (10 mg/kg, i.m.; Zoetis Deutschland GmbH, Berling, Germany), a peripheral venous catheter was established and an initial bolus of propofol (1.5 mg/kg, i.v.; Fresenius Kabi, Bad Homburg Germany) and buprenorphine (0.02 mg/kg, i.v.; Bayer Vital GmbH Tiergesundheit, Leverkusen, Germany) was administered. Subsequently pigs were intubated and mechanically ventilated (Dräger Primus system; Dräger, Lübeck, Germany). Anesthesia was perpetuated with propofol (4–8 mg/kg/h, i.v.) and isoflurane (0.5–2 vol.%; Baxter Deutschland GmbH, Heidelberg, Germany). Of note, no isoflurane was administered before completion of EP measurements to avoid interference of volatile anesthetics with cardiac K2P-channel function (Putzke et al., 2007a (link)).
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