Comparison of Threshold and Tolerance Nociceptive Withdrawal Reflexes in Horses

Simple Summary Nociception is the physiological basis of the complex experience of pain. An established model for its quantification in equine studies is based on the nociceptive withdrawal reflex evoked by electrical stimulation of a sensory nerve. The reflex is recorded via electromyography and it is common to determine the threshold at which a nociceptive-specific reflex activity can be observed. In the present study, the classical methodology was expanded for a deeper understanding of the physiology of nociceptive reflexes in horses. First, for each individual horse, a threshold was determined as the minimal stimulation intensity able to evoke a nociceptive withdrawal reflex. Second, the stimulation intensity was stepwise increased up to tolerance, which was defined as the stimulus that is able to elicit the maximal tolerable behavioral reaction. The characteristics of the reflex activity on the electromyographic records were compared for threshold and tolerance stimulation intensities. At tolerance, the reflex became faster and wider than at threshold, indicating that either a spinal summation mechanism or the recruitment of faster sensory fibers occurs in response to high-intensity noxious stimuli. A novel endpoint (i.e., tolerance) can now be considered when applying the nociceptive withdrawal reflex model in equine studies. The nociceptive withdrawal reflex (NWR) is used to investigate nociception in horses. The NWR threshold is a classical model endpoint. The aims of this study were to determine NWR tolerance and to compare threshold and tolerance reflexes in horses. In 12 horses, the NWR was evoked through electrical stimulation of the digital nerve and recorded via electromyography from the deltoid. Behavioral reactions were scored from 0 to 5 (tolerance). First, the individual NWR threshold was defined, then stimulation intensity was increased to tolerance. The median NWR threshold was 7.0 mA, whereas NWR tolerance was 10.7 mA. Upon visual inspection of the records, two main reflex components R1 (median latency 44 ms) and R2 (median latency 81 ms) were identified at threshold. Increasing stimulation intensity to tolerance led to a significant increase in the amplitude and duration of R1 and R2, whereas their latency decreased. At tolerance, a single burst of early, high-amplitude reflex activity, with a median latency of 39 ms, was detected in 15 out of 23 stimulations (65%). The results of this study suggest that (1) it is feasible to determin