For the neurogenesis and cell fate study, all animals were sacrificed and perfused at PD 28. A telazol:ketamine:xylazine solution was administered intramuscularly at 4.4 mg/kg BW (50.0 mg of tiletamine, plus 50.0 mg of zolazepam, reconstituted with 2.50 mL ketamine [100 g/L] and 2.50 mL xylazine [100 g/L]; Fort Dodge Animal Health, Fort Dodge, IA, USA). Eye blink and pain reflexes were tested to confirm deep anesthesia before piglets were perfused transcardially with phosphate buffered saline (PBS) and a 4% paraformaldehyde/PBS solution. Brains were extracted and post-fixed overnight. The hippocampus was removed and transferred to PBS with 30% sucrose until the tissue sank (~2 days). The entire hippocampus was sectioned using a cryostat into 40 μm sections in an axial plane from dorsal to ventral and collected into a 12-well plate. Six sections were collected into each well with a distance of 480 μm separating each well. As there is variation along the dorsal-ventral axis, tissue from the dorsal region of the hippocampus was used for staining to maintain consistency.
Tiletamine
Manufactured by Zoetis
Sourced in United States
Tiletamine is a veterinary anesthetic agent manufactured by Zoetis. It is a dissociative anesthetic that induces a trance-like state, providing analgesia and immobilization. The core function of Tiletamine is to facilitate safe and effective anesthesia in veterinary procedures.
Automatically generated - may contain errors
4 protocols using tiletamine
1
Piglet Hippocampus Perfusion and Sectioning
2
Landrace Pig Anesthesia Protocol
3
Porcine Fecal and Tissue Collection
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Fecal samples were collected utilizing a fecal loop on PND 31 (CONT, n = 6; ID, n = 7) and/or PND 60 (N = 20; n = 10 per phase 1 diet), snap frozen in liquid nitrogen, and stored at −80°C until processing. At PND 32 (CONT, n = 6; ID, n = 7) and PND 61 or 62 (N = 20; n = 10 per phase 1 diet), pigs were euthanized to allow tissue collection. All animals were euthanized in a food-deprived state, with access only to water for at least 6 h prior to euthanasia. Pigs were anesthetized using an intramuscular injection of telazol:ketamine:xylazine administered at 0.022 mL/kg bodyweight (50.0 mg tiletamine plus 50.0 mg of zolazepam reconstituted with 2.50 mL ketamine (100 g/L) and 2.50 mL xylazine (100 g/L); Fort Dodge Animal Health, Overland Park, KS). Pigs were euthanized using a 390 mg/mL sodium pentobarbital solution (Patterson Veterinary Supply, Columbus, OH) at 1 mL/5 kg body weight with an intracardiac injection. On the day of tissue collection, AC and fecal samples were collected after euthanasia at PND 32 or PND 61, snap frozen in liquid nitrogen, and stored at −80°C until processing.
4
Rhesus Macaque Blood Collection for Assays
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25 Rhesus macaques were sedated with 2mg/kg of 50mg/ml 1:1 tiletamine/zolazepam intramuscularly (Fort Dodge Animal Health, Fort Dodge, IA, USA) and baseline whole blood collected via femoral blind stick in vacutainers using EDTA as an anti-coagulant during routine quarantine procedures (BD Biosciences, San Jose, CA, USA). Samples were assigned a numerical value for blinding purposes and paired for use in both NOX capacity and whole blood stimulation assays.
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