Mice were anesthetized with 7 μL/g of body weight of a mixture of Xylazine (Rompun 2%; Bayer Health Care, Leverkusen, Germany) and Ketamin (Ketavet; Zoetis, Berlin, Germany) in 0.9% NaCl, immobilized on a stereotactic frame (David Kopf Instruments, Tujunga, CA, USA) in flat-skull position and kept warm. A midline incision was made with a scalpel. One by ten [5 (link)] cells/μL in medium without supplements were implanted by stereotactic injection 1 mm anterior and 1.5 mm right to the bregma with a 22-gauge Hamilton syringe (Hamilton, Bonaduz, Switzerland) after drilling a whole into the skull with a 23G needle. At a depth of 4 mm, cells were slowly injected within 2 min and, after a settling period of another minute, the needle was removed in 1 mm step/minute. The incision was sutured and patched with Opsite spray dressing (Smith&Nephew, London, UK).
Ketavet
Manufactured by Zoetis
Sourced in Germany, United Kingdom
Ketavet is a pharmaceutical product manufactured by Zoetis. It is a sterile injectable solution that contains the active ingredient ketamine hydrochloride. Ketavet is primarily used as a general anesthetic and analgesic in veterinary medicine.
Automatically generated - may contain errors
Lab products found in correlation
Rompun 2, by Bayer (4 mentions)
Rompun, by Bayer (4 mentions)
Stereotactic frame, by Kopf Instruments (2 mentions)
Ketamine hydrochloride, by Zoetis (2 mentions)
Butorphanol tartrate, by MSD (1 mentions)
Domitor, by Vetoquinol (1 mentions)
Nanoject 2, by Drummond (1 mentions)
Spectralis hra oct device, by Heidelberg Engineering (1 mentions)
Bepanthen, by Bayer (1 mentions)
13 protocols using ketavet
1
Stereotactic Implantation of Tumor Cells in Mice
2
Badger Anesthesia for Capture and Handling
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The badger is anaesthetised, in the trap where it was captured, by intramuscular injection in the thigh region of a triple combination of ketamine hydrochloride (100 mg ml–1, Ketavet, Zoetis UK Ltd), medetomidine hydrochloride (1 mg ml–1, Domitor, Vetoquinol UK Ltd) and butorphanol tartrate (10 mg ml–1, Dolerex, MSD Animal Health UK Ltd) at a ratio of 2:1:2 by volume, respectively, and a dose rate of approximately 0.2 ml kg–1 (equivalent to 8 mg kg–1 ketamine hydrochloride, 0.04 mg kg–1 medetomidine hydrochloride and 0.8 mg kg–1 butorphanol tartrate) (de Leeuw et al.2004 (link)) (each badger’s bodyweight is estimated by visual assessment).
3
VEGF-A Intraocular Injection in Neonatal Mice
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Intraocular injection of human recombinant VEGF-A165 (Reliatech, 300-076) was performed in P6 pups that were anesthesized by intraperitoneal injection of xylazine (Bayer, Rompun 2%; 10 mg kg−1) and ketamine (Zoetis, Ketavet 100 mg ml−1; 100 mg kg−1) dissolved in saline. The eyelids were carefully separated using a scalpel and 0.5 µl of VEGF-A165 at a concentration of 5 μg μl−1 were injected into the vitreous humor using glass capillary pipettes with a micromanipulator (Nanoject II, Drummond Scientific). Pups were kept under controlled conditions for 2 h before EdU injection. Samples were collected 2 h after EdU administration (i.e., 4 h after VEGF-A165 injection).
4
Optic Nerve Crush and CaspNPs Injection
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Unilateral optic nerve crush surgery was performed using a custom-made forceps (Sautter and Sabel, 1993). Briefly, the rats were anesthetized by intraperitoneal injection of Ketavet (0.75 mL/kg) (ketamine hydrochloride; Zoetis Deutschland GmbH, Berlin, Germany) and Dormitor (0.5 mL/kg) (medetomidine hydrochloride; Orion Corporation, Espoo, Finland). The optic nerve crush was made with calibrated cross-action forceps (KLS Martin Group, Tuttlingen, Germany) applied for 30 seconds at 2–3 mm from the eye with the forceps jaws spaced 0.2 mm apart to produce a moderate crush. Initially, connective tissue was detached from the sclera and a sharp needle (0.8 mm diameter) was used to puncture it, followed by insertion of a Hamilton syringe with blunt cannula for injection into the vitreous. For the ex vivo study, 3 μL of CaspNPs were injected into the vitreous body of both sides (n = 3) or, for control purposes, blank NPs (loaded with PBS) (n = 3), non-silencing NPs (n = 3), caspase-3 siRNA-Ca2+ particles (n = 3), or PBS (n = 3). For the in vivo study, bilateral ONC was performed, and immediately thereafter, the left eye was treated with 3 μL of CaspNPs and the right eye was treated with 3 μL blank NPs suspended in phosphate buffered saline (PBS, pH 7.4, 0.1 M) as control.
5
Retrograde RGC Labeling in Rat
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One week before retina baseline imaging, RGCs were selectively labeled by retrograde axonal transport of fluorescent tracer (n = 16 rats). To this end, rats were anaesthetized by intraperitoneal injection of Ketavet/Dormitor, Ketavet (0.75 mL/kg) (ketamine hydrochloride; Zoetis Deutschland GmbH, Berlin, Germany) and Dormitor (0.5 mL/kg) (medetomidine hydrochloride; Orion Corporation, Espoo, Finland). After drilling openings in the skull, carboxylate-modified microspheres were stereotactically injected (0.04 μm large FluoSpheres; Ex 580/ Em 605, Invitrogen GmbH, Karlsruhe, Germany) into the right and left superior colliculus (coordinates: 6.9 mm posterior to Bregma and –/+1.2 mm lateral from midline) based on its expected stereotaxic coordinates (Paxinos and Watson, 1998). Slow injections of 0.5 μL were made at each of four different depths below dura (2.5, 3.5, 3.0, 4.0 mm), i.e. 2 μL on each side and allowing 30 seconds for diffusion at each position.
6
Orthotopic GBM Tumor Model in Mice
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Mice received i.p. 7 µL/g body weight of a mixture of 0.1% xylazine (Rompun 2%; Bayer, Leverkusen, Germany) and 1.5% ketamine (Ketavet; Zoetis, Berlin, Germany) in 0.9% NaCl. A middle incision was made on the skin with a scalpel after disinfection with a 10% povidone iodine solution. To prevent the animals’ corneas from drying out, their eyes were covered with Bepanthen cream. Mice were immobilized on a stereotactic frame in a flat-skull position. After drilling a hole into the skull with a 23 G needle tip (coordinates 1.0 m anterior and 1.5 mm right of the bregma), 1 μL of cells (1 × 105 murine GBM cells/μL or 5 × 104 human GBM cells/μL in a supplement-free medium) was slowly injected within two minutes with a 22 G Hamilton syringe at a depth of 3 mm (the syringe was vertically inserted 4 mm and retracted 1 mm). Finally, the syringe was retracted 1 mm/min, and the skin was carefully sutured. For microglia tracing, Cx3cr1::creER2, R26-RFP mice received i.p. 75 mg/kg/d tamoxifen (dissolved in corn oil) in an intraperitoneal injection. The tamoxifen injection was performed three consecutive days. For in vivo depletion of GBM-TK cells, GCV was applied i.p. on four consecutive days from 14 to 17 DPO at 50 mg/kg/d.
7
Unilateral Viral Vector Injection in Mouse SNc
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Mice were deeply anesthetized with a mixture of Ketavet® (100 mg/kg, Zoetis Deutschland GmbH; Berlin, Germany) and 2% Rompun® (5 mg/kg, Bayer; Leverkusen, Germany), diluted in 0.9% NaCl. Later mice were placed in a stereotactic frame (Kopf Instruments, Tujunga, CA) for operation [37] . A total amount of 2µl rAAV5-CBA-human-α-synuclein (wild-type; original titer: 1x10 13 vg/ml) or rAAV5-CBA-luciferase (original titer: 1x10 13 vg/ml) was unilaterally injected into right side of the SNc over ten minutes (flow rate of 200 nl/min) by microinjection pump (Micro4TM, WPI, Sarasota, USA). After the injection, the needle stayed in the brain for an additional 10 minutes before it was slowly retracted. The injections were performed by using a microsyringe, stainless steel needle (33G, WPI, Sarasota, USA). The coordinates of the injection were anteroposterior: -3.1 mm, mediolateral: -1.2 mm, and dorsoventral: -4.2 mm, relative to bregma using a flat skull position [38] .
The viral-mediated vectors were provided by the Michael J. Fox Foundation, and the Gene Therapy Center of the University of North Carolina at Chapel Hill, USA.
The viral-mediated vectors were provided by the Michael J. Fox Foundation, and the Gene Therapy Center of the University of North Carolina at Chapel Hill, USA.
8
Quantifying Retinal Thickness in Mice
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For OCT, mice were anesthetized by intraperitoneal injection of ketamine hydrochloride (100 mg/kg body weight, Ketavet; Zoetis) and xylazine hydrochloride (10 mg/kg body weight, Rompun; Bayer HealthCare) diluted in 0.9% sodium chloride. The pupils of the mice were dilated using 2.5% phenylephrine and 0.5% tropicamide before OCT. Spectral-domain optical coherence tomography (SD-OCT) was performed on both eyes with a Spectralis™ HRA/OCT device (Heidelberg Engineering) to quantify the retinal thickness using the Heidelberg Eye Explorer Software with circular ring scans (circle diameter 3 and 6 mm), centered around the optic nerve head.
9
Orthotopic Glioblastoma Xenograft Model
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Mice received i.p. 7 µL/g body weight of a mixture of 0.1% xylazine (Rompun 2%; Bayer, Leverkusen, Germany) and 1.5% ketamine (Ketavet; Zoetis, Berlin, Germany) in 0.9% NaCl. Their corneas were covered with a moisturizing cream (Bepanthen; Bayer). After disinfection with 10% potassium iodide solution, a midline incision was made in the skin of the skull with a scalpel. Mice were immobilized on a stereotactic frame (David Kopf Instruments, Tujunga, CA, USA). After drilling a hole into the skull with a 23G needle tip at the puncture point 1.5 mm anterior and 1.5 mm right of the bregma, 1 μL PBS or GBM cells (1 × 105 Gl261 or human GBM cells/µL and 5 × 104 murine GBM cells/µL) was slowly injected into the mouse brain at a 3 mm depth within 2 min using a 22G Hamilton syringe (Hamilton, Bonaduz, Switzerland). Afterwards, the syringe was removed in 1 mm steps per minute and the skin was sutured. Tumors were grown for 3 weeks for murine cells and for up to 6 weeks for human GBM cells, as previously reported [24 (link)].
10
Primate Welfare in Animal Research
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Monkeys were housed at the German Primate Center under conditions approved in
accordance with §§ 7–9 of the German Animal Welfare Act and European
Union guidelines (EU directives 86/609/EWG and 2010/63/EU). An external
ethics committee of the Lower Saxony State Office for Consumer Protection and
Food Safety (LAVES) approved the experiments from which animals were analyzed
for this retrospective study. Monkeys were constantly monitored by
veterinarians and animal caretakers, and a scoring system with end point
guidelines was applied upon development of clinical symptoms. The animals
were euthanized through an overdose of pentobarbital
(Narcoren®, Merial, Hallbergmoos, Germany)
after anesthesia with a combination of ketamine
(Ketavet®, Zoetis, Berlin, Germany),
xylazine (Rompun®, Bayer Vital GmbH,
Leverkusen, Germany), and atropine (Atropine Sulfate B.
Braun®, B. Braun Melsungen AG, Melsungen,
Germany).
accordance with §§ 7–9 of the German Animal Welfare Act and European
Union guidelines (EU directives 86/609/EWG and 2010/63/EU). An external
ethics committee of the Lower Saxony State Office for Consumer Protection and
Food Safety (LAVES) approved the experiments from which animals were analyzed
for this retrospective study. Monkeys were constantly monitored by
veterinarians and animal caretakers, and a scoring system with end point
guidelines was applied upon development of clinical symptoms. The animals
were euthanized through an overdose of pentobarbital
(Narcoren®, Merial, Hallbergmoos, Germany)
after anesthesia with a combination of ketamine
(Ketavet®, Zoetis, Berlin, Germany),
xylazine (Rompun®, Bayer Vital GmbH,
Leverkusen, Germany), and atropine (Atropine Sulfate B.
Braun®, B. Braun Melsungen AG, Melsungen,
Germany).
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