However, it remains to be seen whether the presence of inflammation in the face would affect the quality of the “grimace”. Also, this method is only valuable for pain of short-to-moderate duration and would not be useful for chronic studies. Cytoskeletal Signaling inhibitor stimulus-evoked behavior testing methods The whisker pad region of rodents is a tricky area to study stimulus-evoked behavior. This region has a rich mechanosensory receptor sheet, which is stimulated in nearly continuous haptic activities during exploratory behavior, and these complex whisker movements can complicate the testing. On the other hand, the IoN is a large and relatively easily Inhibitors,research,lifescience,medical accessible sensory-only nerve,

and innervates a large area which has been the region of choice for many studies. When studying stimulus-evoked behavior in the orofacial region, one of the major pitfalls is the criterion of the “response”. Inhibitors,research,lifescience,medical In the paw region, a reflex-like withdrawal of the paw from the stimulation source is usually considered as the response. In the facial region, the responses may vary from scratching and blinking to grimaces and removing the head. All possible responses need to be classified before testing and analyzing. Vos et al. (1994) have

set a standard for orofacial pain testing in the first report of IoN-CCI in rats. Inhibitors,research,lifescience,medical They have thoroughly studied the rat’s behavior following the CCI intervention, including spontaneous activity (face grooming, exploratory behavior) and evoked Inhibitors,research,lifescience,medical behavior which included stimulation with various thicknesses of von Frey filaments and a pin prick. Based on the responses, a “response score” was attributed, combining the various criteria. We have recently adapted a simplified version of such quantification in mice, where face-grooming behaviors, withdrawal and aggressiveness toward the probe have been totaled to achieve a response score (Krzyzanowska Inhibitors,research,lifescience,medical et al. 2011). Apart from the challenges of approaching the testing probes to the area of interest, the facial region is tricky to stimulate as rodents tend to actively move their heads, which is especially pronounced in mice.

In addition, mice are particularly active and escape when the stimulating object (such as a von Frey hair) is approached. Rats, on the other hand, are much calmer and it is possible to perform stimulations with von Frey hairs, as demonstrated in numerous publications (Vos et al. 1994; Idanpaan-Heikkila ADP ribosylation factor and Guilbaud 1999; Deseure et al. 2002; Martin and Avendano 2009; Martin et al. 2010). In mice, to date, only a few publications have reported the use of von Frey hairs in the orofacial region. Recently, in a study involving partial IoN ligation, the authors have behaviorally tested mice placed on a mesh floor, restricted within a 8-cm-diameter plastic cup from the top, and stimulated by von Frey hairs from underneath (Xu et al. 2008; Aita et al. 2010).