Three-week-old CD1 strain male mice (Charles Rivers) were used for the studies. Mice were housed in a light-and a temperature-controlled room with ad libitum access to diet and water. Fe-deficient diet (TD 80396, 3/kg Fe, and Fe-replete diet TD 80396, 48 mg/kg ferric citrate (Harlan Teklad) were used for the experiment and the diets composition, a modification of AIN-76A is given by Chaudhury et al. (11) .
Td 80396
Manufactured by Inotiv
Sourced in United States
The TD.80396 is a piece of laboratory equipment. It is used for performing specific tasks or functions within a laboratory setting. However, a detailed description of its core function while maintaining an unbiased and factual approach is not available at this time.
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8 protocols using td 80396
1
Dietary Iron Deficiency in Mice
2
Serum and Liver Iron Overload Mice Models
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All animals were maintained in a pathogen-free, temperature-controlled environment under a 12-h light/dark cycle at Beijing Friendship Hospital. All animal protocols were approved by the Institutional Animal Care and Ethics Committee, and all work was performed under the ethical guidelines of the Ethics Committee of Beijing Friendship Hospital, Capital Medical University (No. 20-2035). The serum iron overload mouse model was established as previously described,8 (link),13 (link) with some modifications. Briefly, 6-week-old male C57BL/6 mice (3–4 per treatment) were intraperitoneally injected with 10 mg of human Holo-Tf in 200 µL of saline or saline alone and were analyzed after 2, 4, and 6 h. A mouse model of liver iron overload was established as previously described,14 (link) with some modifications. Briefly, 6-week-old male C57BL/6 mice (5 per treatment) were fed a high-iron diet (2% carbonyl iron; TD.08496; Envigo, Indianapolis, IN, USA) for 1 week followed by a low-iron diet (2–6 ppm iron; TD.80396; Envigo) for 2 days before sacrifice. Mice that received a standard rodent diet were used as controls.
3
Iron Deficiency Induces Brain ID in Rats
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All experiments were approved by the University of Minnesota Institutional Animal Care and Use Committee. Brain ID was induced in rat pups using an iron-deficient diet regimen described previously [31 (link)]. In brief, timed-pregnant Sprague–Dawley dams (Charles River, Wilmington, MA, USA) were randomly assigned to either iron-deficient (2–6 ppm iron, TD.80396, Envigo, Indiana, IN, USA) or iron-sufficient (200 ppm iron, TD.09256, Envigo, Indianapolis, IN, USA) fortified diets ad libitum from gestational day 3 until postnatal day 7 (P7). Shortly after birth, litters were culled to 8 pups containing at least 2 females. On P7, all dams were placed on the iron-sufficient diet. On P15, pups were sacrificed, and hippocampi were micro-dissected, flash-frozen in liquid nitrogen, and stored at −80 °C until use.
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Dietary Iron Deficiency and Neurodevelopment
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Hepatic PCBP1 Knockout Mice on Iron Diets
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Hepatocyte-Specific PCBP1 Depletion in Mice
7
Fetal-Neonatal Iron Deficiency Protocol
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The University of Minnesota Institutional Animal Care and Use Committee approved all experiments in this study. Gestational day 2 (G2) pregnant Sprague-Dawley rats were purchased from Charles Rivers (Wilmington, MA). Rats were kept in a 12 h:12 h light:dark cycle with ad lib food and water. Fetal-neonatal iron deficiency was induced by dietary manipulation as described previously [32 (link)]. In brief, pregnant dams were given a purified ID diet (4 mg Fe/kg, TD 80396, Harlan Teklad, Madison, WI) from G2 to P7, at which time nursing dams were given a purified iron sufficient (IS) diet (200 mg Fe/kg, TD 01583, Harlan Teklad) to generate ID pups. Both ID and IS diets were similar in all contents with the exception of iron (ferric citrate) content. Control IS pups were generated from pregnant dams maintained on a purified IS diet. All litters were culled to eight pups with six males and two females at birth. Only male offspring were used in experiments.
8
Dietary Iron Deficiency in Mice
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Twelve male CD1 mice (3 wk of age) were made Fe deficient through use of a low-iron diet of ∼3-mg Fe/Kg diet based on the modified AIN-76A purified rodent diet (24 (link)) (TD.80396; Harlan Teklad) for 3 wk (i.e., until they were 6 wk of age). Four mice were also placed on a normal iron-sufficient diet (48-mg Fe/Kg diet) (TD.80394; Harlan Teklad) to serve as control. The diets were of identical composition except that the iron-sufficient diet contained iron added as ferric citrate. After this, blood was withdrawn from the tails to determine the initial Hb concentrations of the mice. The Fe-deficient mice were then divided into 3 treatment groups based on similar Hb concentrations. These 12 mice were maintained on the low-Fe diet in groups of 4, of which 1 group did not receive any iron supplementation (low-iron diet); the 2 other groups were gavaged daily with 150-μg Fe as nano Fe(III) compound or FeSO4 for 7 d (until mice were 7 wk of age). After the 7 d, mice were weighed, anesthetized, and blood samples were taken for Hb and serum iron determinations. The mice were then killed by isoflurane anesthesia followed by neck dislocation, and the spleen, duodenum, kidney, and liver samples were excised, snap frozen in liquid nitrogen, and stored at −80°C until further analysis.
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