‘Optical Von-Frey’ method to determine nociceptive thresholds: a novel paradigm for preclinical pain assessment and analgesic screening

Abstract

ABSTRACTThe goal of this study was to characterize a model that specifically activates peripheral nociceptors, allowing pure nociceptive thresholds to be monitored over a range of conditions including pathology or in screening analgesic compounds. Transgenic mice expressing channelrhodopsin-2 (ChR2) in cell populations positive for the transient receptor potential cation channel subfamily V member 1 (TRPV1) gene were bred to enable peripheral nociceptor photostimulation. Preliminary experiments confirmed the expected localisation pattern of ChR2 positive profiles in the dorsal root ganglion and superficial dorsal horn, mirroring TPRV1 expression. Brief hindpaw photostimulation with 470nm light caused hindpaw withdrawal and nocifensive behaviours in ChR2 positive animals but not control ChR2 negative animals. Using a simplified up/down approach, ‘optical’ nociceptive thresholds were assessed with a 5-intensity hindpaw photostimulation paradigm, establishing the minimum intensity required to produce a withdrawal response (optical threshold). All testing was also video recorded and analysed post-hoc to assess additional photostimulation evoked behaviours. Repeated testing over several days showed optical nociceptive thresholds and response duration were similar, supporting the stability of these variables across a timeframe relevant to onset of pathology or drug administration. Optical nociceptive thresholds were also assessed following morphine administration (30 mg/kg), which significantly raised thresholds, highlighting analgesic screening utility of this model. Together, these findings demonstrate the peripheral photostimulation with optical thresholding is a useful addition to the preclinical nociception assessment toolkit, with the key advantage of inducing a purely nociceptive response to a non-invasive, non-tissue damaging stimulus.