Ret 3 rectal temperature probe

Hyperthermia-induced seizure experiments were conducted in P20-28 Scn1a +/-mice and wildtype littermates at age P18-25 in order to prime animals to have more frequent spontaneous seizures (Hawkins et al., 2017) . A rodent temperature regulator (TCAT-2DF, Physitemp Instruments, Inc, Clifton, NJ) connected to a heat lamp and RET-3 rectal temperature probe was used. The rectal probe was inserted, and mice acclimated to the temperature probe for 5 min before the hyperthermia protocol was started. Mouse core body temperature was elevated 0.5°C every 2 min until the onset of the first clonic convulsion with loss of posture or until 42.5°C was reached. Mice that reached 42.5°C were held at temperature for 3 min. Spontaneous behavioral seizures were assessed 24-72 h after hyperthermia protocol by an observer blinded to genotype. Animals were brought to an isolated room in the lab and kept in their home cage with ad libitum access to food and water during the daylight cycle. After witnessing a tonic-clonic convulsive seizure, the animal that had the seizure and a wildtype littermate were killed one hour after the event. To capture baseline c-fos levels, mice were handled and acclimated each day in an isolated room in the lab for three days, after which the animals were perfused as previously discussed.