WT and KO mice obtained from SLC48A1 HET crosses were weaned at 21 days of age (P21) and placed on their respective diets, supplemented with deionized water. Standard rodent diet was obtained from Envigo and the two ppm (TD.09127) Fe diet was custom ordered from Envigo, Madison, WI. Body weights and feed intake per cage were measured weekly to ensure the conditions of the animals. Blood was collected by retro-orbital bleeding using microcapillary tubes (Fisher Scientific, cat. number 22–362566). At the end of five weeks, mice were sacrificed by cardiac perfusion using Dulbecco's phosphate-buffered saline (DPBS) (Gibco, cat. number 14190250) under anesthesia (10% ketamine, 8% xylazine mix). Prior to perfusion, whole blood was collected into tubes and allowed to clot at room temperature for 45 min and serum was separated from the sample by centrifugation at 2000 g for 10 min. For the prolonged dietary iron study where the Kaplan Meier survival curve was obtained, whole blood hematocrits were obtained every two weeks until week 14. All surviving KO mice were sacrificed when the survival rate fell below 50% (~week 16), while WT mice were kept on the diets until week 18.
Standard rodent diet
Manufactured by Inotiv
The Standard Rodent Diet is a complete and balanced feed formulated to meet the nutritional requirements of rodents. It provides essential vitamins, minerals, and macronutrients necessary for maintaining the health and well-being of laboratory rodents.
Automatically generated - may contain errors
Lab products found in correlation
Two ppm td 09127 fe diet, by Inotiv (2 mentions)
Microcapillary tubes, by Thermo Fisher Scientific (2 mentions)
Dulbecco s phosphate buffered saline dpbs, by Thermo Fisher Scientific (2 mentions)
Brown norway rat, by Charles River Laboratories (1 mentions)
Female sprague dawley rats, by Charles River Laboratories (1 mentions)
5 protocols using standard rodent diet
1
Dietary Iron Deficiency in Mice
2
Dietary Iron Deficiency in Mice
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WT and KO mice obtained from SLC48A1 HET crosses were weaned at 21 days of age (P21) and placed on their respective diets, supplemented with deionized water. Standard rodent diet was obtained from Envigo and the two ppm (TD.09127) Fe diet was custom ordered from Envigo, Madison, WI. Body weights and feed intake per cage were measured weekly to ensure the conditions of the animals. Blood was collected by retro-orbital bleeding using microcapillary tubes (Fisher Scientific, cat. number 22–362566). At the end of five weeks, mice were sacrificed by cardiac perfusion using Dulbecco's phosphate-buffered saline (DPBS) (Gibco, cat. number 14190250) under anesthesia (10% ketamine, 8% xylazine mix). Prior to perfusion, whole blood was collected into tubes and allowed to clot at room temperature for 45 min and serum was separated from the sample by centrifugation at 2000 g for 10 min. For the prolonged dietary iron study where the Kaplan Meier survival curve was obtained, whole blood hematocrits were obtained every two weeks until week 14. All surviving KO mice were sacrificed when the survival rate fell below 50% (~week 16), while WT mice were kept on the diets until week 18.
3
Aged Brown Norway Rat Housing Protocol
4
Sprague-Dawley Rat Breeding and Housing
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Female Sprague‐Dawley rats used for generating control and PNS offspring were purchased from Charles River (Margate, UK). All rats were group‐housed in individually ventilated cages (IVCs), under a 12:12 hour light/dark photocycle (lights on at 08.00 am ) at 22 ± 1˚C and 58 ± 3% relative humidity, and with access to drinking water and standard rodent diet (Harlan Teklad) available ad lib., unless otherwise stated. Breeding females were fed a 50:50 mixture of 14% and 19% protein diet (Harlan Teklad) throughout pregnancy and lactation. All animal experiments were approved by the local Animal Welfare and Ethical Review Body and performed in accordance with the UK Animals (Scientific Procedures) Act 1986 and the European Directive (2010/63/EU).
5
Evaluating Brain Water Content in Rats
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All animal protocols were approved by the Johns Hopkins Institutional Animal Care and Use Committee and were carried out in male Sprague-Dawley rats (Charles River Laboratories, Inc., Wilmington, DE, USA). Before being used for the experiments, animals were fed with a standard rodent diet (Teklad, Madison, WI, U.S.A.) and were housed at 24.1°C and 30% humidity with a 11:00 A.M.– 11:00 P.M. light-dark cycle. Spontaneously breathing rats were initially anesthetized by isoflurane (1–2%) in an oxygen-air mixture delivered through a face mask. Then, after being weighed, they were sacrificed by exsanguination from a cardiotomy performed under deep isoflurane anesthesia. During exsanguination, the head was elevated to ensure elimination of blood from the brain. The whole brain, including olfactory bulbs, cerebellum, and brainstem, was procured, weighed, dried at 35–38°C for 2 days, and weighed again. Brain water content was calculated as follows: % H2O = (1 –dry weight/wet weight) x 100 [11 (link)]. A total of 129 animals were studied in 12 groups. Each group was of a different average age and weight when ordered from the supplier, ranged in number from 2 to 20 animals, and did not overlap in age with other groups. The median [interquartile range (IQR)] group size was 9 [8 (link), 12 (link)] rats.
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