Four-week-old male C57BL/6J mice were purchased from Harlan Laboratory (Houston, TX, USA). All animal experiments were approved by the Institutional Animal Care and Use Committee of Texas A&M University and were performed in compliance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research. Mice were housed under temperature- and humidity-controlled, 12L:12D cycle conditions. All mice were given food and water ad libitum. At 5 weeks of age (body weight, 20 g), mice were fed a standard laboratory chow (control diet: 10% fat calories, 20% protein calories, and 70% carbohydrate calories; Research Diets, Inc., New Brunswick, NJ, USA) or an HFD (59.4% fat calories, 18.1% protein calories, and 22.5% carbohydrate calories; TestDiet, St. Louis, MO, USA). After 2 months of HFD regimen, half of the HFD mice were given daily metformin treatments at a dose of 150 mg/kg through oral gavage. Body weight and food intake were measured weekly. Nonfasting blood glucose levels and glucose tolerance were measured monthly by taking blood from the tail vein. Glucose levels were measured using Clarity Plus blood glucose monitoring system (Diagnostic Test Group, Boca Raton, FL, USA).
High fat diet (hfd)
Manufactured by TestDiet
Sourced in United States
The HFD is a laboratory equipment used for the preparation and analysis of high-fat diets. It is designed to accurately measure and mix the various components required for formulating high-fat dietary compositions for research purposes. The core function of the HFD is to provide a consistent and controlled environment for the development and study of high-fat diets.
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10 protocols using high fat diet (hfd)
1
High-Fat Diet and Metformin Treatment in Mice
2
Exercise Interventions for High-Fat Diet-Induced Obesity
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Mice were fed a high-fat diet (HFD; 58Y1, TestDiet, Richmond, IN, USA) and were administered the intervention for 4, 8, and 16 weeks. The SED group received no intervention. Swimming was selected as an exercise intervention because mice are natural and self-motivated swimmers [29 (link)]. Mice were subjected to swimming without workload in tanks containing water at 31 ± 1 °C for 2 h a day, 5 days a week [30 (link),31 (link)]. The WBV group were exposed to vibrations on a vertically oscillating platform (BW-760, BodyGreen, Taipei, Taiwan). During WBV, the mice were temporarily housed in one of the six compartments of an acrylic cage fixed to the top of the platform. The WBV stimulus was applied for 30 min a day, 5 days a week, at a frequency of 13 Hz, and at an acceleration of 0.68 g (1 g = 9.81 m/s2) and the maximum amplitude of 2 mm [32 (link)].
3
Inducing Insulin Resistance and Lipid Sensing Defects
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The 3-day HFD model has previously been shown to induce hepatic and hypothalamic insulin resistance and upper small intestinal lipid-sensing defects39 (link),42 (link)–44 (link),49 (link). The day following surgery, rats were given free access to a 10% lard-enriched HFD (Test Diet, St. Louis, MO, USA). Food intake was monitored daily and rats that were hyperphagic and consumed more calories than rats receiving regular chow were included in the study.
4
Dietary Obesity and Depression Mechanisms
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Male C57BL/6N mice (8-week-old) were randomly assigned to regular chow diet (CD) and HFD groups with computer-based randomization. HFD mice were fed with commercial HFD (Cat# 58Y1, TestDiet, St. Louis, MO, USA) for 12 weeks to induce obesity, systemic insulin resistance, and depressive phenotypes. Five cohorts of mice were subjected to the following studies: 1) examination of the effects of HFD on the exhibition of depression-like behaviors and activities of glutamatergic afferents to the NAc; 2) identification of the neural circuit that determines the depression-like behavior in HFD mice using the chemogenetic approach; 3 and 4) clarifications of the role of glial glutamate transporters in the developments of circuit maladaptations and behavioral deficits by lentiviral knockdown and overexpression approaches; 5) examination of the therapeutic effect of glutamate modulator, riluzole (RLZ), on HFD-induced depression-like behaviors.
5
Transgenic Murine Model for CEACAM1 Study
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This study was conducted under protocol number 08017 and was approved by the Institutional Animal Care and Use Committee of the City of Hope (Association for Assessment and Accreditation of Laboratory Animal Care [AAALAC] assurance number A3001‐01). The HFD was from TestDiet (catalog #1810740). Mice were housed in a specific pathogen‐free facility, four per cage, and were fed ad libitum. Human CEACAM1 transgenic mice (huTg) were generated on the FVB/NJ mice background (13 (link)) and backcrossed over seven generations to the Ceacam1−/− background C57/BL6 mice in our lab. Because the homozygous huTg mice were sterile, only heterozygous huTg mice were used in this study. The number of animals per group (total of 92), parameters measured, and outcomes for each study are shown in the figures and figure legends.
6
HFSTZ-induced Metabolic Dysfunction in Mice
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HFSTZ has been shown to induce metabolic stresses such as hyperglycemia, obesity, insulin resistance, and glucose intolerance [19 –21 (link)]. Male C57BL/6J wild-type (WT) siblings and APPswe/PS1dE9 transgenic (AD) mice were fed with a normal chow diet (NCD, MF-18, Oriental Yeast Co. Ltd., Tokyo, Japan) with water ad libitum. At the age of 10 weeks, half of WT and AD mice randomly chosen were fed with an HFD (60% energy from fat, TestDiet, St. Louis, MO, USA). After 2 weeks, HFD-fed mice were intraperitoneally injected with 50 mg/kg STZ as the HFSTZ induction group. NCD-fed mice were injected with vehicle (0.1 M citrate buffer, pH 4.5). Four experimental groups including NCD WT, NCD AD, HFSTZ WT, and HFSTZ AD mice were sacrificed after 11 weeks of dietary manipulations. The average weight of NCD WT, NCD AD, HFSTZ WT, and HFSTZ AD mice prior to the dietary manipulations were not significantly different (24.70 ± 0.69, 23.85 ± 0.93, 26.07 ± 0.65, and 25.73 ± 0.62, respectively).
7
HFSTZ Mice Model for Metabolic Disorders
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HFSTZ Mice were created by feeding a high-fat diet and injecting single-dose STZ [3 (link),4 (link),5 (link)]. Briefly, 10 week-old mice were fed a HFD (60% energy from fat; TestDiet, St. Louis, MO, USA) with water ad libitum. For control, a normal chow diet (NCD; MF-18, Oriental Yeast Co. Ltd., Tokyo, Japan) was used. Mice on HFD also received intraperitoneal injections of STZ (50 mg/kg, in 0.1 M citrate buffer, pH 4.5) 2 weeks after HFD initiated (HFSTZ group). On the contrary, NCD mice were injected with vehicle (0.1 M citrate buffer, pH 4.5). The HFD continued for 11 weeks of HFD manipulation. To evaluate the effect of APS on ameliorating the impairment in HFSTZ mice, APS (500 mg/kg) was orally administrated twice per day for 7 weeks after 4 weeks of HFD administration. The body weight and blood glucose were recorded every week after dietary manipulations except the week for STZ injection.
8
High-Fat Diet Induced Obesity in Rats
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Twelve-week-old male Wistar rats (body weight 350 to 400 g) were kept in a temperature-controlled room (25 ± 1°C) with 12 h light-dark cycle (lights on at 06:00). The rats were given water and food ad libitum and were divided into two groups according to the diet. One group was fed with standard laboratory diet (3.04 kcal/g). The other group was fed with HFD (5.16 kcal/g) (TestDiet, Richmond, IN, USA) for 6 months. The study protocol was approved by institutional animal care and use committee. All animal-handling procedures were performed according to the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health, as well as the guidelines of the Animal Welfare Act.
9
Genetic Mouse Models for Metabolic Research
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C57BL/6, Rag1-/- and Tbx21-/- (formal gene name Tbx21) mice were purchased from The Jackson Laboratory (Bar Harbor, ME) Rag1-Tbet DKO mice were obtained through crossbreeding of Rag1−/− mice with Tbx21−/− mice. Genotyping for Rag1-Tbet DKO was performed by Transnetyx. Mice were fed either standard chow (normal diet, ND) or a lard-based high fat diet (HFD). The ND, consisting of 4.09 kcal/gram,13.4% kJ/fat, was purchased from Lab Diet (St. Louis, MO). The HFD, consisting of 5.10 kcal/gram, 60% kJ/fat, was purchased from TestDiet (St. Louis, MO). ND was started at week four of life and maintained for 12-15 weeks for the ND group. For the HFD group experiment, HFD feeding was started between weeks five and seven of life. HFD continued for 12 weeks. All mice were bred and maintained within a pathogen-free facility in the Division of Comparative Medicine at Georgetown University Medical Center, with a standard 12-hour light-dark cycle. All procedures on animal subjects were fully approved by the Georgetown University Institutional Animal Care and Use Committee (protocol #2016-1351).
10
Gut Microbiome Dynamics in Trisomic Mice
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All mice were individually housed in multi-take metabolism cages (PheCOMP, Panlab-Harvard Instruments, Barcelona, Spain) and provided with standard chow (SC) and water for two weeks to allow habituation to the housing conditions. After habituation, mice were assigned to each diet-condition. These mice served for the assessment of the genotype-dependent differences in feeding behavior and microbiome composition in a nutritionally balanced diet condition. To investigate the effect of obesogenic high-fat diet (HFD) of microbiome composition in trisomic mice, all mice were fed free-choice HFD and faecal samples were collected before the introduction of HFD and after 2, 14 and 28 days of HFD. To obtain information about the gut microbial profile of trisomic mice we sequenced the: 16S rRNA gene sequences from 6 Ts65Dn and 6 WT mice; and the total RNA from 2 Ts65Dn and 3 WT mice, to validate the 16S results. The mice fed HFD had free access to HFD pellets (HFD, Test Diet, USA) and SC (SC SDS, UK). For the 16S rRNA sequencing the faecal samples were collected under standard chow diet (SC SDSand after 28 days of HFD, while for the total RNA sequencing after 2 and 14 days of HFD (day 14: WT n=2). Pellets were renewed at least twice a week to ensure the maintenance of their organoleptic properties.
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