Generation of CaMKIId-Deficient Lepr db/db Mice CaMKIId-KO mice (12) were crossed to Lepr db/db mice (The Jackson Laboratory) to obtain CaMKIId-deficient Lepr db/db (Lepr db/db /KO) and Lepr 1/1 (Lepr 1/1 /KO) compared with Lepr db/db /wild-type (WT) and Lepr 1/1 /KO. These mice were maintained in a C57BL/6J genetic background, were fed a standard diet, and were kept on a 12-h light and dark cycle at 22 6 2°C and room humidity of 55%. All experimental procedures were reviewed and approved by the Regierungspräsidium Institutional Animal Care and Use Committee, Karlsruhe, Germany.
Leprdb db
Manufactured by Jackson ImmunoResearch
Sourced in Montenegro
Leprdb/db is a mouse strain that carries a mutation in the leptin receptor gene, which leads to obesity, diabetes, and other metabolic disorders. This strain is commonly used in research to study the underlying mechanisms of these conditions.
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Lab products found in correlation
C57bl 6j, by Jackson ImmunoResearch (5 mentions)
Lepob ob, by Jackson ImmunoResearch (3 mentions)
Wild type c57bl 6, by Jackson ImmunoResearch (2 mentions)
Balb c, by Jackson ImmunoResearch (2 mentions)
C57bl 6, by Charles River Laboratories (2 mentions)
B6 sjl, by Jackson ImmunoResearch (1 mentions)
Ot 2, by Jackson ImmunoResearch (1 mentions)
C57blks j, by Jackson ImmunoResearch (1 mentions)
Glucometer elite, by Bayer (1 mentions)
D12492, by Research Diets (1 mentions)
C57bl 6, by Research Diets (1 mentions)
Lfd d 12450b, by Research Diets (1 mentions)
C57bl 6j, by Research Diets (1 mentions)
Loxtbmc4r, by Jackson ImmunoResearch (1 mentions)
Lepr db db mice, by Jackson ImmunoResearch (1 mentions)
Amln diet, by Research Diets (1 mentions)
F0173, by Bio-Serv (1 mentions)
D09100301, by Research Diets (1 mentions)
Td 06414, by Inotiv (1 mentions)
Thbs1 mice, by Jackson ImmunoResearch (1 mentions)
11 protocols using leprdb db
1
Generation of CaMKIId-Deficient Lepr db/db Mice
2
Notch1 Knockout Mice Generation
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The Notch1lox/lox and Cd4-Cre::Notch1lox/lox (Notch1-/-) strains were a gift from Freddy Radtke (Wolfer et al., 2001 (link)). Foxp3tm4(YFP/Cre)Ayr/J, C57BL/6J, B6SJL, OT-II and Leprdb/db strains were obtained from the Jackson Laboratory. Notch1lox/lox and Foxp3tm4(YFP/Cre)Ayr/J strains were crossed to generate Foxp3-Cre::Notch1lox/lox mice. Notch1lox/lox mutant mouse stains were backcrossed with C57BL/6 mice. Except where specified, all experiments used mice within the age group of 8–12 weeks. Mice were housed in controlled temperature and light environments that are maintained in high barrier conditions with specific IVC (individually ventilated cages) controlled systems. The housing environment is tested and routinely monitored for the full pathogen panel recommended by FELESA (Federation of Laboratory Animal Science Associations). Breeding colonies were maintained in-house and all experimental protocols were approved by the Institutional Animal Ethics Committee (NCBS-AEC-AS-6/1/2012; INS-IAE-2016/01[N]) and are in compliance with the norms of the Committee for the Purpose of Control and Supervision of Experiments on Animals, Govt. of India.
3
Animal Procedures in Metabolic Disease
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All animal procedures were performed according to protocols approved by the Institutional Animal Care and Use Committee and in agreement with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Animals were housed in AALAC approved animal facilities of the Cleveland Clinic. Wild type C57BL6, BALB/c, Lepr db/db, Dock7 m + /Lepr db (heterozygote control, lean and normoglycemic) were purchased from The Jackson Laboratories.
Animals were sacrificed at end point by exsanguination under anesthesia with ketamine/xylazine (100 mg/15 mg/kg), and organs were collected. Mice used for cell isolation were euthanized by CO2 asphyxiation followed by the cervical dislocation.
Animals were sacrificed at end point by exsanguination under anesthesia with ketamine/xylazine (100 mg/15 mg/kg), and organs were collected. Mice used for cell isolation were euthanized by CO2 asphyxiation followed by the cervical dislocation.
4
Aldosterone-Induced Cardiomyocyte Changes in Diabetic Mice
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Twenty‐four adult (10‐week‐old, both sexes) Leprdb/db (stock #000697) and 24 corresponding wild‐type (WT) mice on C57BL6/J background were obtained from Jackson Laboratory. The animals were kept at standard temperature, humidity, and lighting. Food (Teklad, 2018) and drinking water were provided ad libitum. Osmotic minipumps (Alzet, model 2004) that delivered a continuous infusion of either d‐aldosterone (0.3 μg/h)21 or vehicle (saline with 5% ethanol) for 4 weeks were implanted subcutaneously in 12‐week‐old mice (Figure 1A ). We used block randomization with a block size of 4 animals (for 1 genotype and treatment group in 1 sex), with 16 control (WT+vehicle) and 16 two‐hit (db/db+Aldo) mice included (allowing for detailed isolated myocyte studies), and for the one‐hit controls we used 8 WT+Aldo and 8 db/db+vehicle mice. For proper allocation concealment, animals were recruited blinded based on sequential ear tag numbers randomly assigned by the animal housing facility. Each treatment group included an equal number of male and female animals. Enzymatic isolation of left ventricular (LV) cardiomyocytes was performed as previously described.26
5
Diabetic Nephropathy Study in db/db Mice
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Six 8-week-old male db/db (Lepr db/db) mice (36.5–38.9g) and six nondiabetic control male db/m mice C57BLKs/J (Jackson Laboratories, Bar Harbor, ME) (18.6–20.6g) were purchased from Nanjing Model Animal Center, Jiangsu, China. All animals were housed at proper room temperature (26°C) and humidity (70%) under a controlled light/dark cycle and had free access to water. All animals should be adaptively fed.46 (link) The db/m mice and the db/db mice were fed with standard diet (12% fat, 28% protein and 60% carbohydrates). Blood glucose was randomly measured with Glucometer Elite (Bayer) after 6 h of fasting. Then, DN can be diagnosed when the urine albumin creatinine ratio (ACR) of the db/db mice was over 50 μg/mg and random blood glucose of the db/db mice was over 16.7 mM.46 (link) After collecting the data of blood glucose and 24-hour urine for 4 weeks, animals were anesthetized and killed, then the renal tissues were dissected for the following experiments, respectively. All animal experiments were guided in accordance with the guidelines of the Care and Use of Laboratory Animals by the National Institutes of Health, and all efforts were made to minimize animal suffering.
6
Metabolic Phenotyping of Fam13a Knockout Mice
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All animal procedures were approved by Augusta University’s Institutional Animal Care and Use Committee (IACUC). Mice were maintained under standard conditions with controlled 12 h/12 h-light–dark cycle and 21 ± 1 °C room temperature. Lepob/ob, Leprdb/db and C57BL/6 J mice were obtained from Jackson Laboratory (Bar Harbor, ME). Fam13a−/− mice were backcrossed to C57BL/6 (Stock#: 027, Charles River) for seven generations and genotyped as previously reported [14 (link)]. 4–5 week old Fam13a+/+ and Fam13a−/− littermates were fed with high fat diet (HFD, D12492, 60% Kcal from fat, Research Diets, NJ) for up to 12 weeks. Mice were not randomized. Six week old C57BL/6 J male mice were simply randomized into two experimental groups for ad libitum access to low fat control diet (LFD, D12450B, 10% Kcal from fat, Research Diets, NJ) and 60% HFD for up to 12 weeks as reported [16 (link)]. All mice studies were not blinded. All mice were sacrificed after 4 h fasting with isoflurane.
7
Obesity and Metabolic Phenotypes in Mice
8
Leptin-receptor-deficient Diabetic Mouse Model
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Male C57BLKS/J homozygous Leprdb/db mice and their heterozygous leptin-receptor-deficient nondiabetic lean littermate controls Leprdb/+ were obtained from Jackson’s Laboratories (Sacramento, CA, USA) and housed at the Primate Unit and Delft Animal Centre (PUDAC) of the South African Medical Research Council (SAMRC) in a controlled environment with a 12 h light/dark cycle in a temperature range of 23–25 °C (relative humidity: ~50%). The mice received standard laboratory chow pellets (Afresh Vention, Cape Town, South Africa ) ad libitum and had free access to drinking water. The study was performed according to principles and guidelines of the South African Medical Research Council’s Guidelines on Ethics for Medical Research: Use of Animals in Research and Training, 2004 (http://ww.mrc.ac.za/ethics/ethicsbook3.pdf ) under the institutional ethical approval of the SAMRC (ECRA No. 07/13) as well as Stellenbosch University Ethics Committee (SU-ACUM13-00021).
9
Mouse Acclimation and Housing
10
Obesity and NASH Mouse Models
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12 week old C57Bl/6J (wild-type, WT) mice, leptin-receptor deficient mice (Lepr db/db ), and 16week-old mice with diet-induced obesity (DIO) were purchased from Jackson Laboratories. Mice were fed normal chow (5V5R, Lab Supply), dustless pellet diet (F0173, Bio-Serv), 12 weeks of high fat diet (60% calories from fat, TD.06414, Envigo Teklad), or 24 weeks of NASH-high fat diet (AMLN-diet, D09100301, Research Diets), where indicated. Prior to all interventions, mice were standardized across groups ensuring equal distribution of body weight for the various treatment groups.
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