For sodium appetite feeding studies (Figure S6G,H ), male Hsd11b2-Cre mice aged 8–16 weeks were acclimated to a two-diet choice of high sodium diet (1.6% Na+ / 4% NaCl; Envigo TD.92034) or standard diet (0.2% Na+ / 0.49% NaCl; Envigo TD.96208) for at least 5 days before commencing food intake experiments in the home cage. Because the high sodium diet formulation differed slightly from our in-house standard chow, these experiments used the appropriate control diet matching its formulation offered by the manufacturer. For dark-cycle feeding experiments, the two diets were pre-weighed and male Hsd11b2-Cre mice received a vehicle or CNO injection approximately 10 minutes prior to the onset of dark. Food was again weighed each hour for the next 3 hours and the remaining food weight values were subtracted from the initial pre-weighed food to provide the amount eaten. In water restriction experiments, animals had ad libitum access to high sodium and standard diets and were acclimated to three-hour restricted H2O access (11:00 – 14:00) for two days prior to the experiment. Next, diets were pre-weighed and animals received vehicle or CNO injections 10 minutes prior to H2O access. Food intake was measured for the next three hours.
Td 96208
Manufactured by Inotiv
The TD.96208 is a piece of laboratory equipment. It is a general-purpose instrument used for various applications in a research or laboratory setting. The core function of the TD.96208 is to perform specific tasks or measurements, but a detailed description without interpretation or extrapolation is not available.
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4 protocols using td 96208
1
Sodium Appetite Feeding Studies
2
Plasma Renin Activity Measurement in Olfr558 Mice
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Plasma renin activity was measured in 3-month-old Olfr558 WT and KO mice with a modified angiotensin I measurement kit (S-1188, Peninsula Laboratories). Plasma was collected from male and female Olfr558 WT and KO mice treated with 0.49% NaCl diet (catalog no. TD.96208 modified with orange color, Envigo). From males, we also collected plasma from mice given Teklad diet (Teklad 2018SX, 18% protein). Plasma was diluted 15-fold and then incubated with excess porcine angiotensinogen (SCP0021, Sigma-Aldrich) for 20 min at 37°C in a buffer containing 50 mM sodium acetate (pH 6.5), 10 mM 4-(2-aminoethyl) benzenesulfony fluoride hydrochloride (Sigma-Aldrich), 10 mM EDTA (pH 8.0), 1 μM porcine angiotensinogen, and 10 mM 8-hydroxyquinoline (Sigma-Aldrich). After incubation, the sample was analyzed according to the provided protocol. Plasma renin activity was assayed by competitive binding of angiotensin I antibody.
3
Rodent Models for Cardiovascular Research
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Male and female (16–20 weeks of age) Sprague Dawley rats from Envigo (Indianapolis, IN) were used. Rats used in intramedullary infusion experiments were maintained on our standard 7917 Irradiated NIH‐31 Mouse/Rat diet (0.8% NaCl; Envigo). Rats used in telemetry experiments were maintained on TD 96208 (0.49% NaCl [Envigo], matched control diet for our high salt diet, TD 92034, 4% NaCl [Envigo]). Male and female wild‐type and GPER knockout mice (18–25 weeks of age) were generated and genotyped as previously described.22 , 23 Mice were maintained on 5053 Picolab Rodent diet 20 (0.82% NaCl; LabDiet, St. Louis, MO). Animals were conventionally housed in a temperature (18°C–23°C) controlled room with a 12:12‐hour light‐dark cycle with free access to food and water. All protocols were in accordance with the Guide for the Care and Use of Laboratory Animals and were approved in advance by the University of Alabama at Birmingham or Tulane University Institutional Animal Care and Use Committee.
4
Sprague Dawley Rats: Age-Matched Dietary Sodium Study
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Male and female age‐matched (13–15 weeks old) Sprague Dawley rats were purchased from Envigo (Indianapolis, IN). Rats were maintained on an NS diet (0.49% NaCl, TD 96208; Envigo) for 2 weeks. Then, a nutrient‐matched HS diet (4% NaCl, TD 92034; Envigo) was introduced for 5 consecutive days. During the entire experimental period, animals were housed in a temperature‐controlled room (22–24 °C) with a 12:12‐hour light‐dark cycle, with free access to water. Experiments were conducted over 4 sequential days to ensure that different stages of the estrus cycle were presented in female rats. All animal protocols were in accordance with the Guide for the Care and Use of Laboratory Animals and were approved in advance by the University of Alabama at Birmingham Institutional Animal Care and Use Committee.
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