Indirect calorimetry of mice was measured using the metabolic cages (Harvard Apparatus) from the Behavioral Analysis Unit at the Institute of Neurobiology, UNAM. Animals were housed at 22°C on a 12 h/12 h light/dark cycle with free access to food and water. Respiratory quotient (RQ) and energy expenditure (EE) were measured for two days in the fourth week of treatment with sulpiride.
Metabolic cage
Manufactured by Harvard Apparatus
Sourced in United States
About the product
Metabolic cages are laboratory equipment designed to study the metabolic activities of small animals, such as rodents. These cages allow for the collection and measurement of various parameters, including food and water intake, urine, and feces production. The cages are designed to maintain the animal's natural environment while providing researchers with the necessary data to analyze the subject's metabolic processes.
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39 protocols using metabolic cage
1
Metabolic effects of sulpiride in mice
2
Optimization of Mercuric Chloride Exposure in Guinea Pigs
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We purchased 4-week-old male Hartley guinea pigs from Charles River Laboratories (Wilmington, MA, USA). We fed them a normal diet (No. 5025, LabDiet, St. Louis, MO, USA) and provided water ad libitum in an environmentally controlled room with a 12:12-h light-dark cycle and the temperature set at 23 °C. All experiments were conducted in accordance with the Guide for the Care and Use of Laboratory Animals of the United States Department of Agriculture, the Vanderbilt University Institutional Animal Care and Use Committee, and the U.S. Army Medical Research and Development Command Animal Care and Use Review Office.
Guinea pigs were kept for 7 days prior to any treatment to recover from the stress of conveyance and to acclimate to their new housing environment. Then, we anesthetized them with isoflurane and implanted a sterile silicone catheter (0.5 mm inner diameter, 0.94 mm outer diameter) into the right jugular vein. The free end of the catheter was passed subcutaneously to the back of the neck, where it was fixed and occluded with a metal plug following a flush of heparinized saline (200 U heparin/mL saline) [90 ]. After the procedure, animals were housed individually in a metabolic cage (Harvard Apparatus, Holliston, MA, USA) for 7 days to recover from the stress associated with the surgery before we began the studies for optimization of mercuric chloride dose and length of exposure and for measuring the toxicant-induced changes in gene expression.
Guinea pigs were kept for 7 days prior to any treatment to recover from the stress of conveyance and to acclimate to their new housing environment. Then, we anesthetized them with isoflurane and implanted a sterile silicone catheter (0.5 mm inner diameter, 0.94 mm outer diameter) into the right jugular vein. The free end of the catheter was passed subcutaneously to the back of the neck, where it was fixed and occluded with a metal plug following a flush of heparinized saline (200 U heparin/mL saline) [90 ]. After the procedure, animals were housed individually in a metabolic cage (Harvard Apparatus, Holliston, MA, USA) for 7 days to recover from the stress associated with the surgery before we began the studies for optimization of mercuric chloride dose and length of exposure and for measuring the toxicant-induced changes in gene expression.
3
Energy Metabolism and Body Fat Analysis
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Mice were subjected to energy metabolism analysis using a metabolic cage (Harvard Apparatus, Holliston, MA, USA) and body fat content analysis with a magnetic resonance imaging (MRI) system 3.0T (Siemens, Munich, Germany) at the Peking University Health Science Center.
4
Urine Collection from Mice Exposed to PG:13C-VG Aerosols
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Prior to exposures, mice were held and a small drop
ofd -glucose:saccharin solution (3.0%/0.125% w/w; Sigma-Aldrich;
St.
Louis, MO) was touched to their mouth. For the 13C-VG study,
we mixed PG (1.0 mL), VG (0.8 mL), and 13C-VG (0.2 mL)
for a final 50:50 (PG:VG) ratio and exposed mice to aerosols for 6
h. After 6 h exposures (air or PG:13C-VG), mice were placed
singly per metabolic cage (Harvard Apparatus; Cambridge, MA) for urine
collection without food yet with access to glucose:saccharin drinking
water. Urine was collected in graduated cylinders surrounded by 4
°C water-jacketed organ baths from 0 to 3 h post exposure, as
well as in a second overnight collection (3–16+ h, O/N) during
which mice were provided both glucose/saccharin solution and food.36 (link) Urine samples were centrifuged (1800g, 5 min; to pellet feces or food) before being decanted
and stored at −80 °C.
of
St.
Louis, MO) was touched to their mouth. For the 13C-VG study,
we mixed PG (1.0 mL), VG (0.8 mL), and 13C-VG (0.2 mL)
for a final 50:50 (PG:VG) ratio and exposed mice to aerosols for 6
h. After 6 h exposures (air or PG:13C-VG), mice were placed
singly per metabolic cage (Harvard Apparatus; Cambridge, MA) for urine
collection without food yet with access to glucose:saccharin drinking
water. Urine was collected in graduated cylinders surrounded by 4
°C water-jacketed organ baths from 0 to 3 h post exposure, as
well as in a second overnight collection (3–16+ h, O/N) during
which mice were provided both glucose/saccharin solution and food.36 (link) Urine samples were centrifuged (1800g, 5 min; to pellet feces or food) before being decanted
and stored at −80 °C.
5
Creatinine Clearance Measurement in Rats
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Urine samples were collected at the fifth week of the experiment by housing the rats in metabolic cages (Harvard Apparatus; Holliston, MA, USA) for 24 h. Urine volumes were recorded and blood samples were collected from the lateral tail vein. Urine and blood samples were centrifuged at 3500 rpm for 15 min and immediately stored at − 80 °C. Serum and urine creatinine levels were measured using a creatinine assay kit (Sigma-Aldrich). The creatinine clearance rate (Ccr) was used to estimate the glomerular filtration rate (GFR) and calculated using the standard formula: urinary flow rate × urine creatinine level/serum creatinine level.
6
Quantifying Acrolein Exposure and Metabolism
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Immediately prior to HEPA‐ and charcoal‐filtered air exposure or CAP exposure, mice were weighed and briefly exposed to D‐glucose/saccharin solution (w/v; 3.0%/0.125%; Sigma) on the mouth (Conklin, Haberzettl, Lesgards, et al., 2009 ; Wood et al., 2001 (link)). After 6h air or CAP exposure, mice were placed singly per metabolic cage (Harvard Apparatus) with D‐glucose/saccharin solution to collect urine (in graduated cylinders inside 4°C water‐jacketed organ baths) for 3 h. After urine collection, mice were placed in home cages overnight with food and water per normal housing arrangements. Urine samples were centrifuged (600 × g, 5 min, 4°C; to pellet any feces and food particles), decanted, and stored at −80°C. The major metabolite of acrolein, 3‐hydroxypropylmercapturic acid (3HPMA), was quantified in urine by mass spectrometry (negative ion mode) as previously described (Conklin et al., 2017 (link)). To account for urinary dilution, values for urinary 3HPMA were normalized to urinary creatinine (mg/dL).
7
Aging-Related Changes in Cardiovascular and Renal Function
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Tail systolic blood pressure (SBP) and heart rate (HR) were recorded monthly by using tail-cuff plethysmography in unanesthetized rats (LE 5001-pressure meter, Letica SA, Barcelona, Spain) from 2 to 8 months old. Each animal was placed in a heater at 37°C for 10 min and placed in a warm electric carpet to maintain vasodilation during determinations. SBP and HR were measured at least 10 times per rat waiting for a period of 2-3 min between each measurement. The mean value of the last 3 determinations that achieved a difference <5 mm Hg was taken as SBP and HR for each animal. All rats were weighed and housed monthly in metabolic cages (Panlab, Barcelona, and Spain) with free access to food and drinking water for a 24-h period during which food and fluid intakes were measured and 24-h urine samples collected. Urine samples were centrifuged for 15 min at 1,000 g and frozen at -80°C until assay. We measured monthly urine volume, proteinuria, AlaAp, GluAp, and DPP4 urinary activities. Urinary excretion of Klotho was measured at 2, 5, and 8 months old. These variables were expressed per mg of urinary creatinine. DOI: 10.1159/000516843
After completion of the experimental period (8 months old), rats were anaesthetized with ketamine (80 mg/kg)/xylazine (15 mg/kg) i.p. Blood samples were then drawn by abdominal aortic puncture to determine plasma variables. Blood samples were centrifuged for 15 min at 1,000 g and stored at -80°. Finally, the rats were killed by cervical dislocation and kidneys and heart were quickly removed and weighed. Plasma variables were: urea, creatinine, AlaAp, GluAp, DPP4, and Klotho. The tibial length was measured to normalize the heart and kidney weight, since body weight cannot be used for this purpose in the present experimental setting. One kidney was harvested without perfusion, fixed in 10% neutral buffered formaldehyde solution for 48 h and subsequently placed in 70% ethanol for histological studies.
After completion of the experimental period (8 months old), rats were anaesthetized with ketamine (80 mg/kg)/xylazine (15 mg/kg) i.p. Blood samples were then drawn by abdominal aortic puncture to determine plasma variables. Blood samples were centrifuged for 15 min at 1,000 g and stored at -80°. Finally, the rats were killed by cervical dislocation and kidneys and heart were quickly removed and weighed. Plasma variables were: urea, creatinine, AlaAp, GluAp, DPP4, and Klotho. The tibial length was measured to normalize the heart and kidney weight, since body weight cannot be used for this purpose in the present experimental setting. One kidney was harvested without perfusion, fixed in 10% neutral buffered formaldehyde solution for 48 h and subsequently placed in 70% ethanol for histological studies.
8
Metabolic Cage Assessment of Injury Response
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One day prior to injury and at the specified end-points (1 day, 2 days, 7 days, 14 days, and 28 days post-injury), animals were placed in metabolic cages (Harvard Apparatus) in which food (grams of chow consumed over 6 h) and water intake, as well as urine output were recorded. All rats were placed in metabolic cages for about 6 h (started approximately at 08:30 and ended at 14:30). After 6 h was over, rats were removed from the metabolic cages and returned to their home cages. Urine, remaining water, and remaining rat chow were measured and recorded. If rats had not yet urinated after 6 h, they were tickled gently to induce urination. Urine was collected to a chilled collection cup placed on ice and centrifuged at 5000 rpm for 5 min at 4°C, to remove any undissolved material, and supernatant was stored at -80°C, until subsequent processing/analyses.
9
Chronic Exposure Metabolic Profiling
10
Metabolic Cage Assessment of Neuroendocrine Function
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