Ret 3 rectal probe

Mice were anesthetized by inhalation of isoflurane (4%, v/v air) for 3 min, and their backs were shaved. Because anesthesia affects thermoregulation, the mice were placed on a heating plate (HP-4M Small Animal Heating Plate, Physitemp) that was connected to a temperature controller (TCAT -2LV; Physitemp), while being kept under anesthesia by isoflurane (<2.5%, v/v air) during the whole subsequent intervention period. A rectal temperature probe (RET-3 Rectal Probe, Physitemp) connected to the temperature controller was inserted into the rectum of mice to measure their core body temperature. Their core body temperature was set to be stable at approx. 36.6°C by automatically switching the heating plate on and off. Approached from the back, both pads of interscapular BAT (left and right) were exposed by a midline incision in the skin and white fat along the upper dorsal surface. Flexible probes (IT-18 Flexible Implantable Microprobe, Physitemp) were plugged into both the left and right BAT pad to monitor the local temperature in BAT. The core body temperature and temperature in BAT were recorded per second during the whole intervention by a sensitive temperature data acquisition system (THERMES-USB; Physitemp) connecting to a laptop using Dasylab software (Version 12.0).

Naïve BALB/c mice fed either HFD to induce obesity or LFD as control were sensitized with PN flour as we have described previously [17 (link)]. In each experiment, mice were split into four groups: LFD PBS, LFD PN, HFD PBS, and HFD PN. In total across two experiments, 5 mice were in each PBS group, 6 mice were in the LFD PN group, and 8 mice were in the HFD PN group. Briefly, mice were exposed to either 100 μg peanut flour in 50 μL PBS or PBS alone twice per week for 4 weeks. On day 27, mice were retroorbitally bled to determine serum levels of PN-specific IgE and IgG1. The next day (day 28), mice were challenged with 2.5 mg PN peanut extract in 500 μL PBS via intraperitoneal injection to induce anaphylactic reaction. Rectal temperature and clinical symptoms were monitored before (0 min) and after PN challenge (every 10 min for 1 h). Rectal temperatures were recorded with an electronic thermometer (Oakton Instruments, Vernon Hills, Ill) equipped with a RET-3 rectal probe (Physitemp Instruments, Clifton, NJ). Clinical symptoms were scored based on the following published criteria [18 (link)]: 0, no symptoms; 1, scratching of ear and mouth; 2, puffiness around eyes and mouth, pilar erection, labored breathing; 3, prolonged period of motionlessness; 4, severely reduced motility, tremors, severe respiratory distress; and 5, death.

F1.Scn1a^+/- mice between P14 to P16 received an intraperitoneal injection of 0, 0.3, 1, or 30 mg/kg doses of clobazam solubilized in vegetable oil (10 ml/kg volume). After 15 minutes, each mouse was placed in a cylindrical container and fitted with a RET-3 rectal probe (Physitemp) connected to a heat lamp via a temperature controller (TCAT-2DF (Physitemp) reconfigured with a Partlow 1160+ controller). At 20 minutes post-injection, mouse body temperature was elevated by 0.5°C every two minutes until a maximum of 42.5°C was reached. When a GTC seizure occurred (rearing and falling with forelimb clonus), mice were removed from the container and temperature was recorded. If after 3 minutes at 42.5°C no seizure occurred, the mouse was removed and recorded as a seizure-free.