Body composition was measured using a Bruker MiniSpec NMR. Whole-body VO2, RER, and activity levels were measured using a CLAMS system (Columbus Instruments). Cold-tolerance testing was carried out in a 4 °C cold room. Prior to cold-tolerance testing, the mice were injected with a temperature-sensitive transponder (Bio Medic Data Systems, IPTT 300). One week after the injections, the mice were transferred to a 4 °C cold room for 6–8 h, and their core temperature was assessed using a Reader-Programmer (Bio Medic Data Systems, DAS 8007). The mice were single-housed in cages containing bedding with access to food and water throughout the cold-tolerance test. For long-term cold-exposure experiments, the mice were acclimated to the cold following a protocol used to acclimate UCP1 null mice to the cold [25 (link)]. Briefly, the mice were housed in a rodent incubator at 18°C for 2 weeks, after which the temperature was lowered to 6.5°C for 7 days. The mice were euthanized after a 4 h fast and their tissues were harvested. For glucose-tolerance testing, their blood glucose levels were measured following an IP injection of 20% glucose (5 μL/g body mass) at time points 0, 15, 30, 60, and 120 min.
Das 8007
Manufactured by Avidity Science
The DAS 8007 is a high-performance digital acquisition system designed for use in scientific and industrial applications. It features multiple analog input channels, high-speed data acquisition, and advanced signal processing capabilities. The core function of the DAS 8007 is to capture, digitize, and process analog signals with high accuracy and precision.
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3 protocols using das 8007
1
Metabolic Phenotyping of Cold Tolerance
2
Intermittent Cold Exposure Experiments
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Cold exposure experiments were performed as described previously44 (link). In brief, subcutaneous, biocompatible, and sterile microchip transponders (IPTT-300; Bio Medic Data Systems, Seaford, DE, USA) were implanted in male and female mice in all groups at least 5 days prior to experimentation. On the day of the experiment, mice were housed singly in pre‐chilled cages at 4 °C with free access to water. Body temperature was assessed every 30–45 min for 6–8 h using a wireless reader system (DAS-8007; Bio Medic Data Systems).
According to our IACUC protocol we defined “survival” when mouse body temperature remained >28 °C. At any temperature equal or below 28 °C, mice were rescued by removal to room temperature. The time of removal was recorded and considered as drop-out.
For experiments in which cell viability was detected in vivo, we applied an intermittent cold exposure which allows for longer-term cold exposure while preventing animal death. Animals were exposed to cold at intervals of 4–6 h intermitted by recovery periods till the mice gained back their core body temperature on a course of 5 days.
According to our IACUC protocol we defined “survival” when mouse body temperature remained >28 °C. At any temperature equal or below 28 °C, mice were rescued by removal to room temperature. The time of removal was recorded and considered as drop-out.
For experiments in which cell viability was detected in vivo, we applied an intermittent cold exposure which allows for longer-term cold exposure while preventing animal death. Animals were exposed to cold at intervals of 4–6 h intermitted by recovery periods till the mice gained back their core body temperature on a course of 5 days.
3
Assessing Cold Tolerance and BAT Activity in Mice
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Cold tolerance testing was conducted on single housed mice at 4°C with access to water and light bedding. Core body temperature was measured using a temperature-sensitive transponder injected into dorsal subcutaneous adipose tissue (Bio Medic Data Systems, IPTT 300). The transponder was placed in the mice 1 week before cold tolerance testing. Transponder readings were assessed using a Reader-Programmer (Bio Medic Data Systems, DAS 8007). Whole-body VO2 was measured using the Comprehensive Lab Animal Monitoring System (Columbus Instruments). Intraperitoneal injected CL 316,243 (Sigma-Aldrich, C5976) was used to stimulate BAT activity.
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