Weight matched mice were acclimated to metabolic cages for 48 hours, and on day 2 were injected with ASO. Thereafter, physical activity, oxygen consumption (VO2), carbon dioxide production (VCO2), and respiratory exchange ratio (RER) were continually monitored for an additional 48 hr using the Oxymax CLAMS system (Columbus Instruments) at a temperature of 22°C. Data represent the last 6 a.m. to 6 a.m. period after adequate acclimation.
Oxymax clams system
Manufactured by Columbus Instruments
Sourced in United States
The Oxymax CLAMS system is a compact, integrated laboratory instrument designed for the measurement of oxygen consumption, carbon dioxide production, and other metabolic parameters in small animals. The system utilizes an open-circuit indirect calorimetry method to provide continuous, high-resolution data on the animal's metabolic activity.
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Lab products found in correlation
25 protocols using oxymax clams system
1
Metabolic Monitoring in Acclimated Mice
2
Metabolic Profiling in Rodents
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Respirometric heat production, respiratory exchange ratio (RER), physical activity by photoelectric beam break, and food intake/patterning were assessed using a 16-chamber OxyMax CLAMS system (Columbus Instruments). Animals were housed individually in specialized OxyMax home cages for four nights in total. Ambient temperature was maintained at 30 ± 1 °C throughout the testing period. Animals were acclimated to the system for 48 hours, and data were recorded and analyzed from the remaining 48 hours of the testing period. Data from 17 minute bins were averaged across days within animal before group comparisons were made. Aerobic heat was calculated by the system using the Lusk equation, as above72 .
3
Metabolic Profiling of Mice
4
Metabolic Analysis of Mice in Chambers
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During week 17, six mice in each group were housed individually in metabolic chambers at 22°C, allowed free access to food and water, and acclimated 24 hours prior to metabolic assessments. Measurements were taken for a 24-h period, including a 12-h light cycle and a 12-h dark cycle. Oxygen consumption (VO2), carbon dioxide production (VCO2), and physical activity (by infra-red beam breaks) were measured every 20 minutes using a computer-controlled, open-circuit Oxymax/CLAMS System (Columbus Instruments, Columbus, OH, USA). Respiratory exchange ratio (RER) was calculated as the ratio of VCO2 to VO2. Heat, the standard measure of energy expenditure, was calculated with the formula [18 (link)],
5
Systemic Energy Metabolism Profiling
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The profile of systemic energy metabolism was determined by indirect calorimetry using the Oxymax CLAMS system (Columbus Instruments) at 7th month post infection. Mice were housed individually in CLAMS cages and allowed to acclimate for 24 h (12 h day and 12 h night) in 25°C with unrestricted excess to food and water. O2 consumption (VO2), CO2 release (VCO2), Respiratory Exchange Ratio (RER) and food intake were recorded for 24 h spanning a single light-dark cycle. Carbohydrate oxidation was calculated using the formula [(4.585∗VCO2)–(3.226∗VO2)]∗4, in which the 4 represents the conversion from mass per time unit to kcal per time unit (Péronnet and Massicotte, 1991 (link)). Similarly, fat oxidation was calculated using the formula [(1.695∗VO2)-(1.701∗VCO2)]∗9 (Péronnet and Massicotte, 1991 (link)).
6
Metabolic Profiling of Mice
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Seven weeks into the dietary protocol, mice were individually placed into metabolic cages at thermoneutrality (28 °C) to determine oxygen consumption (VO2), carbon dioxide production (VCO2), and the respiratory quotient by indirect calorimetry. Animals were initially placed in their individual metabolic cage for 24 h before the beginning of the experiment. Measurements were made continuously during the following 2 days (36 measurements · mouse−1 · day−1) in an Oxymax CLAMS system (Columbus Instruments, Columbus, OH, USA). Data presented are the average of all measurements made over 2 days of monitoring. Body composition was measured by nuclear magnetic resonance with a Bruker Minispec (LF90) apparatus.
7
Respiration and Locomotion Monitoring
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Respiration rates (in breaths per minute) and animal locomotive activity (as ambulation) were assessed using the Oxymax/CLAMS system (Columbus Instruments, Columbus, OH, USA) as described previously [60 (link),63 (link)]. Mice were habituated to their individual sealed housing chambers for 60 min before testing. Mice were administered [Nal(2′)4]CJ-15,208 (100 nmol, i.c.v.), morphine (30 nmol, i.c.v.), or vehicle, as indicated, and 5 min later confined to the CLAMS testing chambers. Pressure monitoring within the sealed chambers measured frequency of respiration. Infrared beams located in the floor measured locomotion as number of beam breaks. Respiration and locomotive data were averaged over 20 min periods for 80 min post-injection of the test compound. Data are presented as % vehicle response ± SEM; ambulation, or breaths per minute.
8
Indirect Calorimetry for Mouse Energy Metabolism
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Mouse energy metabolism was determined by indirect calorimetry using the Oxymax CLAMS system (Columbus Instruments) in the Rodent Behavioral Core of the Cleveland Clinic Lerner Research Institute. Mice were housed individually in CLAMS cages and allowed to acclimate for 48 h with unrestricted excess to food and water. Thereafter, O2 consumption (VO2), CO2 release, RER and heat generation were recorded for 24 h spanning a single light-dark cycle.
9
Comprehensive Assessment of Body Composition and Exercise Capacity in Mice
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Body composition from wild‐type and Tazkd mice at 3 and 6 months of age was measured with dual‐energy x‐ray absorptiometry (DEXA Lunar PIXImus; GE, Madison, WI, USA). Animals were anesthetized with 100 mg·kg−1 ketamine and 10 mg·kg−1 xylazine. For CLAMS, mice were housed individually and acclimatized for 1 day. Oxygen consumption, carbon dioxide release, energy expenditure (EE) and activity were measured using a Columbus Instruments Oxymax‐CLAMS system according to guidelines for measuring energy metabolism in mice [41 , 42 ].
Mice were physically challenged on a lidded motorized treadmill (Columbus Instrument, Columbus, OH, USA), which had adjustable speed and inclination and electric shock stimulation grid. The stimulus intensity was set to 1 mA. Mice were acclimatized in the treadmill for 3 days before the test. Acclimation protocol: 5 m·min−1 for 5 min, followed by 1 m·min−1 increment in speed every min until 10 min. Mice were allowed to rest for 5 min and then run for 10 min at 10 m·min−1. On the test day, wild‐type and tazkd mice were placed in the treadmill and the test started with the mice running for 5 min at 5 m·min−1; then, the speed was increased 1 m·min−1 every min up to 20 min. Exhaustion was achieved when a mouse stayed more than 10 s on the stimulus grid or touched the grid more than 10 consecutive times.
Mice were physically challenged on a lidded motorized treadmill (Columbus Instrument, Columbus, OH, USA), which had adjustable speed and inclination and electric shock stimulation grid. The stimulus intensity was set to 1 mA. Mice were acclimatized in the treadmill for 3 days before the test. Acclimation protocol: 5 m·min−1 for 5 min, followed by 1 m·min−1 increment in speed every min until 10 min. Mice were allowed to rest for 5 min and then run for 10 min at 10 m·min−1. On the test day, wild‐type and tazkd mice were placed in the treadmill and the test started with the mice running for 5 min at 5 m·min−1; then, the speed was increased 1 m·min−1 every min up to 20 min. Exhaustion was achieved when a mouse stayed more than 10 s on the stimulus grid or touched the grid more than 10 consecutive times.
10
Morphine-Induced Respiratory Changes Assessed
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Respiration rates (in breaths per minute) were assessed using the Oxymax/CLAMS system (Columbus Instruments, Columbus, OH, USA) as described previously [27 (link)]. Mice were habituated to their individual sealed housing chambers for 60 min before testing. Mice were administered morphine (20 mg/kg, i.p.) or vehicle, as indicated, and 5 min later, confined to the CLAMS testing chambers. Additional mice received pretreatment with saline (i.n.s.) or OL-CTOP (100 or 600 μg, i.n.s.) 45 min prior to morphine (20 mg/kg, i.p.). Pressure monitoring within the sealed chambers measured the frequency of respiration. Respiration data were averaged over 20 min periods for 120 min post-i.p. injection of the saline or morphine. Data are presented as % vehicle response ± SEM, breaths per minute.
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