An automated method to analyze vocalization of unrestrained rats submitted to noxious electrical stimuli

Unrestrained rats were subjected to electrical stimuli applied to their paws via an electrified cage floor. Intensity, duration, and order of stimulation were chosen after preliminary tests. Vocalization threshold and vocalization as a behavioral response were studied. The vocalization was recorded and the signal analyzed by a simple computerized method that calculated five parameters: delay, maximal amplitude, duration, area, and maximal derivative with respect to time. The last four parameters increased with increasing intensity of stimulation and remained stable when the same stimulation was given repeatedly. Sensitivity to morphine (2.5 and 5 mg/kg s.c.) was tested. Morphine raised the threshold and lowered vocalization parameters, and it was antagonized by naloxone, thus validating the method. The sensitivity of the test and its capacity to separate sensory and affective components of pain are discussed.