Ng peripheral nerve injury, alterations in global DNA methylation are observed in the PFC and amydala but not in the visual cortex or thalamus, b) environmental enrichment reduces both behavioural signs of neuropathic pain and pathological changes in PFC global methylation, and c) PFC global methylation significantly correlates with the severity of mechanical and cold sensitivity. Long-term alterations in DNA methylation could therefore provide a molecular substrate for chronic pain-related alterations in the CNS, forming a “memory trace” for pain in the brain that can be targeted therapeutically.tightly ligated with 6.0 silk (Ethicon) and sectioned distal to the ligation. The 
sural nerve was left intact. Sham surgery involved exposing the nerve without damaging it [13].Behavioral AssessmentAll animals underwent baseline behavioral assessments immediately prior to surgery and no differences were observed between groups (data not shown). The first cohort were then re-assessed six months following nerve injury or sham surgery control (Figures 1 and 2). In the 
environmental study (Figures 3 and 4), the presence of nerve injury-induced hypersensitivity was confirmed three months following surgery when the environmental MedChemExpress Thiazole Orange manipulations were implemented and again two months after environmental change. Mechanical Sensitivity. Calibrated monofilaments (Stoelting Co., Wood Dale, IL) were applied to the plantar surface of the hindpaw and the 50 threshold to withdraw (grams) was calculated as previously described [14]. The stimulus intensity ranged from 0.008 g to 4 g. Cold Sensitivity. A modified version of the acetone drop test [15] was used: total duration of acetone-evoked behaviors (flinching, licking or biting) was measured for 1 minute after acetone (,25 ml) 18055761 was applied to the plantar surface of the hindpaw with the aid of a blunt needle attached to a syringe. Motor Function. The accelerating rotarod assay was used (IITC Life Science Inc., Woodland Hills, CA) with the mouse adapter [16]. The task includes a speed ramp from 0 to 30 rpm over 60 s, followed by an additional 240 s at the maximal speed. Overall Activity. Mice were individually placed individually into the centre of a transparent open field (26626 cm2) in a quiet room illuminated with white light and their spontaneous behavior was videotaped. The floor of the apparatus was Madrasin equally divided into nine squares. The total number of squares visited in a 5 minute period was assessed. An animal must fully enter the square for it to be considered as visited. Since each square is similar in size to an average mouse (,8?0 cm), the number of squares visited serves as a proxy measure for general motor activity. Anxiety-like behavior. The same open field was used with the primary measure being the time spent in the central square during the 5 minute task [17].Materials and Methods AnimalsTwo cohorts of 8?0 week-old male CD1 mice (Charles River, St-Constant, QC, Canada) were used. Animals were housed in ventilated polycarbonate cages and received water and rodent diet (Teklad Rodent Diet 2020X) ad libitum. Animals in the standard environment (Figures 1 2) were housed in groups of 3? with a cardboard hut and cotton nesting material. In contrast, the enriched environment consisted of three mice/cage, a home cage running wheel mounted on a plastic hut (Mouse IglooH with Fast-Trac running wheel, http://www. bio-serv.com), and marbles. In the impoverished environment, each animal was housed singly.Ng peripheral nerve injury, alterations in global DNA methylation are observed in the PFC and amydala but not in the visual cortex or thalamus, b) environmental enrichment reduces both behavioural signs of neuropathic pain and pathological changes in PFC global methylation, and c) PFC global methylation significantly correlates with the severity of mechanical and cold sensitivity. Long-term alterations in DNA methylation could therefore provide a molecular substrate for chronic pain-related alterations in the CNS, forming a “memory trace” for pain in the brain that can be targeted therapeutically.tightly ligated with 6.0 silk (Ethicon) and sectioned distal to the ligation. The sural nerve was left intact. Sham surgery involved exposing the nerve without damaging it [13].Behavioral AssessmentAll animals underwent baseline behavioral assessments immediately prior to surgery and no differences were observed between groups (data not shown). The first cohort were then re-assessed six months following nerve injury or sham surgery control (Figures 1 and 2). In the environmental study (Figures 3 and 4), the presence of nerve injury-induced hypersensitivity was confirmed three months following surgery when the environmental manipulations were implemented and again two months after environmental change. Mechanical Sensitivity. Calibrated monofilaments (Stoelting Co., Wood Dale, IL) were applied to the plantar surface of the hindpaw and the 50 threshold to withdraw (grams) was calculated as previously described [14]. The stimulus intensity ranged from 0.008 g to 4 g. Cold Sensitivity. A modified version of the acetone drop test [15] was used: total duration of acetone-evoked behaviors (flinching, licking or biting) was measured for 1 minute after acetone (,25 ml) 18055761 was applied to the plantar surface of the hindpaw with the aid of a blunt needle attached to a syringe. Motor Function. The accelerating rotarod assay was used (IITC Life Science Inc., Woodland Hills, CA) with the mouse adapter [16]. The task includes a speed ramp from 0 to 30 rpm over 60 s, followed by an additional 240 s at the maximal speed. Overall Activity. Mice were individually placed individually into the centre of a transparent open field (26626 cm2) in a quiet room illuminated with white light and their spontaneous behavior was videotaped. The floor of the apparatus was equally divided into nine squares. The total number of squares visited in a 5 minute period was assessed. An animal must fully enter the square for it to be considered as visited. Since each square is similar in size to an average mouse (,8?0 cm), the number of squares visited serves as a proxy measure for general motor activity. Anxiety-like behavior. The same open field was used with the primary measure being the time spent in the central square during the 5 minute task [17].Materials and Methods AnimalsTwo cohorts of 8?0 week-old male CD1 mice (Charles River, St-Constant, QC, Canada) were used. Animals were housed in ventilated polycarbonate cages and received water and rodent diet (Teklad Rodent Diet 2020X) ad libitum. Animals in the standard environment (Figures 1 2) were housed in groups of 3? with a cardboard hut and cotton nesting material. In contrast, the enriched environment consisted of three mice/cage, a home cage running wheel mounted on a plastic hut (Mouse IglooH with Fast-Trac running wheel, http://www. bio-serv.com), and marbles. In the impoverished environment, each animal was housed singly.