Transcutaneous electrical nerve stimulation at both high and low frequencies activates ventrolateral periaqueductal grey to decrease mechanical hyperalgesia in arthritic rats

Abstract

Transcutaneous electric nerve stimulation (TENS) is widely used for the treatment of pain. TENS produces an opioid-mediated antinociception that utilizes the rostroventromedial medulla (RVM). Similarly, antinociception evoked from the periaqueductal grey (PAG) is opioid-mediated and includes a relay in the RVM. Therefore, we investigated whether the ventrolateral or dorsolateral PAG mediates antinociception produced by TENS in rats. Paw and knee joint mechanical withdrawal thresholds were assessed before and after knee joint inflammation (3% kaolin/carrageenan), and after TENS stimulation (active or sham). Cobalt chloride (CoCl(2); 5 mM) or vehicle was microinjected into the ventrolateral periaqueductal grey (vlPAG) or dorsolateral periaqueductal grey (dlPAG) prior to treatment with TENS. Either high (100 Hz) or low (4 Hz) frequency TENS was then applied to the inflamed knee for 20 min. Active TENS significantly increased withdrawal thresholds of the paw and knee joint in the group microinjected with vehicle when compared to thresholds prior to TENS (P<0.001) or to sham TENS (P<0.001). The increases in withdrawal thresholds normally observed after TENS were prevented by microinjection of CoCl(2) into the vlPAG, but not the dlPAG prior to TENS and were significantly lower than controls treated with TENS (P<0.001). In a separate group of animals, microinjection of CoCl(2) into the vlPAG temporarily reversed the decreased mechanical withdrawal threshold suggesting a role for the vlPAG in the facilitation of joint pain. No significant difference was observed for dlPAG. We hypothesize that the effects of TENS are mediated through the vlPAG that sends projections through the RVM to the spinal cord to produce an opioid-mediated analgesia.

Figures

Fig. 1

Time line for the experiment.

Fig. 2

Microinjection of cobalt chloride (CoCl2) into the vlPAG 24 h after induction of inflammation increased the mechanical withdrawal threshold of the paw. The effect of CoCl2 peaked 30 min after microinjection and lasted through 90 min.

Fig. 3

Schematic coronal sections of the rat brain adapted from Paxinos and Watson (2005) illustrating approximate sites of microinjections into the vlPAG (A) and dlPAG (B). Numbers indicate that distance from the interaural in millimeters. Only rats with injection sites in or immediately adjacent to the vlPAG or dlPAG were included in data analysis. Symbols represent the microinjection sites; filled symbols indicate animals microinjected with CoCl2 and open symbols, vehicle. Animals were stimulated with high frequency TENS (squares), low frequency TENS (circles) or sham TENS (triangles).

Fig. 4

Bar graph representing mechanical withdrawal threshold of the paw from animals microinjected with either CoCl2 or vehicle into the (A) vlPAG and (B) dlPAG. Mechanical withdrawal thresholds are illustrated prior to induction of inflammation (Baseline), before application of TENS, and after microinjection of the vlPAG or dlPAG. Data are represented as mean±SEM. P value 0.05 was considered statistically significant. * Significantly different from baseline time; † significantly different from vehicle control groups.

Fig. 5

Bar graph representing mechanical withdrawal threshold of the knee from animals microinjected with either CoCl2 or vehicle into the (A) vlPAG or (B) dlPAG. Mechanical withdrawal thresholds are illustrated prior to induction of inflammation (Baseline), before application of TENS, and after microinjection of the vlPAG or dlPAG. Data are represented as mean±SEM. P value <0.05 was considered statistically significant. * Significantly different from baseline time; † significantly different from vehicle control groups.