Home Cage Wheel Running Research Articles

Diurnal sex differences in morphine withdrawal revealed by continuous assessment of voluntary home cage wheel running in the rat

Animal studies modeling recreational opioid use show more severe withdrawal symptoms in male compared to female rats, whereas our study modeling opioid use for pain showed a greater withdrawal-induced decrease in wheel running in female rats. The objective of this experiment was to determine whether sex differences in spontaneous morphine withdrawal are caused by differences in assessment method (i.e., wheel running vs. somatic symptoms). Twice daily injections of morphine (5 – 20 mg/kg, s.c.) for 5 days produced a dose and time dependent decrease in wheel running that was greater in male compared to female rats. Termination of morphine administration resulted in an overall decrease in running and a decrease in the amount of running during the dark phase of the light cycle from 95 % to approximately 75 %. In male rats, this decrease in the percent of dark running was caused by a large decrease in dark phase running, whereas female rats had a slightly higher increase in light phase running. Withdrawal also reduced maximal running speed and caused a decrease in body weight that was larger in male than female rats. Withdrawal symptoms were greatest on the day following the last morphine injection, but persisted for all 3 days of assessment. Morphine withdrawal produced a greater decrease in dark phase wheel running and body weight in male rats and a greater increase in light phase running in female rats. Voluntary home cage wheel running provides a continuous measure of opioid withdrawal that is consistent with other measures of opioid withdrawal.

Continuous fentanyl administration and spontaneous withdrawal decreases home cage wheel running in rats with and without hindpaw inflammation

Fentanyl is a potent analgesic with a rapid onset and short half-life that make it a useful treatment for pain and a lethal drug of abuse. The present study used voluntary home cage wheel running to assess the effect of hindpaw inflammation, fentanyl administration, and spontaneous fentanyl withdrawal. Fentanyl (0.32 or 1.0 mg/kg/day) or placebo osmotic pumps were implanted subcutaneously and rats received an intraplantar injection of Complete Freund's Adjuvant (CFA) or saline. Rats with hindpaw inflammation caused by CFA administration were less active than rats injected with saline into the hindpaw. The antinociceptive effect of 0.32 mg/kg/day of fentanyl was evident as a recovery of wheel running in these rats. Administration of 1 mg/kg/day of fentanyl almost completely inhibited wheel running during the first day in rats with and without hindpaw inflammation. Wheel running increased each subsequent day until the pumps were surgically removed after day 3. Withdrawal from 0.32 or 1 mg/kg/day of fentanyl caused a decrease in wheel running that lasted 2 days in rats without hindpaw inflammation. In contrast, withdrawal was only evident following termination of 1 mg/kg/day of fentanyl in rats with hindpaw inflammation. This decrease in running seemed to persist beyond the 3 days of assessment. These data demonstrate that fentanyl can either depress or restore activity depending on the dose and pain condition. Moreover, termination of 3 days of continuous fentanyl administration resulted in a dose and time dependent decrease in wheel running consistent with opioid withdrawal.

Evaluation of tarsal injuries in C57BL/6J male mice.

Tarsal joint abnormalities have been observed in aged male mice on a C57BL background. This joint disease consists of calcaneal displacement, inflammation, and proliferation of cartilage and connective tissue, that can progress to ankylosis of the joint. While tarsal pathology has been described previously in C57BL/6N substrains, as well as in STR/ort and B10.BR strain, no current literature describes this disease occurring in C57BL/6J mice. More importantly the behavioral features that may result from such a change to the joint have yet to be evaluated. This condition was observed in older male mice of the C57BL/6J lineage, around the age of 20 weeks or older, at a frequency of 1% of the population. To assess potential phenotypic sequela, this study sought to evaluate body weight, frailty assessment, home cage wheel running, dynamic weight bearing, and mechanical allodynia with and without the presence of pain relief with morphine. Overall mice with tarsal injuries had significantly higher frailty scores (p< 0.05) and weighed less (p<0.01) compared to unaffected mice. Affected mice had greater overall touch sensitivity (p<0.05) and they placed more weight on their forelimbs (p<0.01) compared to their hind limbs. Lastly, when housed with a running wheel, affected mice ran for a shorter length of time (p<0.01) but tended to run a greater distance within the time they did run (p<0.01) compared to unaffected mice. When tested just after being given morphine, the affected mice performed more similarly to unaffected mice, suggesting there is a pain sensation to this disease process. This highlights the importance of further characterizing inbred mouse mutations, as they may impact research programs or specific study goals.

Low-Dose Δ9-THC Produces Antinociception in Female, But Not Male Rats.

Introduction: The analgesic effects of delta-9-tetrahydrocannabinol (THC), the primary psychoactive compound in cannabis, have been widely promoted. Unfortunately, animal research is limited by the use of high doses and pain-evoked tests. Motor and psychoactive effects of THC may suppress evoked responses in the absence of antinociceptive effects. Materials and Methods: This study overcomes these problems by assessing the antinociceptive effect of low doses of subcutaneous THC on depression of home cage wheel running caused by hindpaw inflammation. Female and male Long-Evans rats were individually housed in a cage with a running wheel. Results: Female rats ran significantly more than male rats. Administration of Complete Freund's Adjuvant into the right hindpaw produced inflammatory pain that significantly depressed wheel running in female and male rats. Administration of a low dose of THC (0.32, but not 0.56 or 1.0 mg/kg) restored wheel running in the hour after administration in female rats. Administration of these doses had no effect on pain-depressed wheel running in male rats. Conclusions: These data are consistent with previous studies showing greater antinociceptive effects of THC in female compared with male rats. These data extend previous findings by showing that low doses of THC can restore pain-depressed behaviors.

Morphine restores and naloxone-precipitated withdrawal depresses wheel running in rats with hindpaw inflammation

Opioids such as morphine are the most effective treatment for pain, but termination of opioid use can produce severe withdrawal symptoms. The present study models this process by using home cage wheel running to assess well-being as a result of pain, morphine analgesia, and opioid withdrawal. Injection of CFA into the right hindpaw caused a dramatic decrease in wheel running and body weight. Implantation of two morphine pellets (75 mg each) resulted in an increase in body weight on Day 1 of administration and a more gradual restoration of wheel running that was only evident during the dark phase of the circadian cycle on Days 3 and 4 of morphine administration. Continuous morphine administration decreased wheel running during the relatively inactive light phase. These findings are consistent with the clinical goal of pain therapeutics to restore normal activity during the day and facilitate sleep at night. Administration of naloxone (1 mg/kg) on Day 5 of morphine administration depressed wheel running for approximately 4 h and caused an increase in wet dog shakes. Naloxone-precipitated changes were no longer evident 6 h after administration. These findings demonstrate that the use of morphine to treat pain does not protect against opioid withdrawal. Moreover, this study provides additional support for the use of home cage wheel running as a method to assess changes in well-being as a result of pain, analgesia, and opioid withdrawal.

Analysis of Formalin‐depressed Wheel Running and its Reversal by Ketoprofen

Traditional pain-evoked tests (e.g., hot plate and tail withdrawal tests) do not mimic the disruptive effects of pain on daily life, a primary outcome measure used for pain in humans. To solve this problem, we demonstrated that pain-depressed home cage wheel running is an objective and clinically relevant measure of the functional consequences of chronic inflammatory pain in the rat. However, few studies have examined the functional consequences of acute pain; thus, the aim of this study was to examine the effects of acute formalin-induced inflammatory pain on function as measured by home cage wheel running. Rats were housed in individual cages containing a computerized running wheel. Rats were allowed free access to the wheel 23 hours/day for 7 days prior to the induction of hindpaw inflammation to acquire stable running levels. Wheel running during the 23 hours immediately prior to induction of inflammation was used as the baseline. Inflammation was induced with an intraplantar injection of formalin (5%, 0.05 mL) into the right hindpaw. The functional effects of a single formalin injection were apparent as indicated by decreased wheel running after the first hour following injection. Interestingly, wheel running also decreased in the 7th hour following formalin injection. In the 7th hour, wheel running was depressed in the absence of mechanical allodynia and spontaneous pain behaviors, suggesting that pain-evoked behaviors and decreased function are independent of each other. A pretreatment of the nonsteroidal anti-inflammatory drug ketoprofen (10 mg/kg) prevented depression of wheel running in both the 1st and 7th hour. These results indicate that continuous monitoring of the functional consequences of pain using home cage wheel running reveal insights into long-term changes in behavior resulting from acute pain, and that early treatment may prevent functional deficits due to acute pain.

Tetrahydrocannabinol (THC) Exacerbates Inflammatory Bowel Disease in Adolescent and Adult Female Rats

Inflammatory Bowel Disease (IBD) is a life-long disorder that often begins between the ages of 15 and 30. Anecdotal reports suggest cannabinoids may be an effective treatment. This study sought to determine whether home cage wheel running is an effective method to assess IBD, and whether Tetrahydrocannabinol (THC), the primary psychoactive compound in cannabis, can restore wheel running depressed by IBD. Adolescent and adult female Sprague-Dawley rats were individually housed in a cage with a running wheel. Rats were injected with trinitrobenzene sulphonic acid (TNBS) into the rectum to induce IBD-like symptoms. One day later, both vehicle and TNBS treated rats were injected with a low dose of THC (0.32 mg/kg, s.c.) or vehicle. Administration of TNBS depressed wheel running in adolescent and adult rats. No antinociceptive effect of THC was evident when administered 1 day after TNBS. In fact, administration of THC prolonged TNBS-induced depression of wheel running for over 5 days in adolescent and adult rats. These results show that home cage wheel running is depressed by TNBS-induced IBD, making it a useful tool to evaluate the behavioral consequences of IBD, and that administration of THC, instead of producing antinociception, exacerbates TNBS-induced IBD. PerspectiveThis article advances research on inflammatory bowel disease in two important ways: 1) Home cage wheel running is a new and sensitive tool to assess the behavioral consequences of IBD in adolescent and adult rats; and 2) Administration of the cannabinoid THC exacerbates the negative behavioral effects of IBD.

'Reinventing the wheel' to advance the development of pain therapeutics.

Chronic pain affects approximately one-third of the population worldwide. The primary goal of animal research is to understand the neural mechanisms underlying pain so better treatments can be developed. Despite an enormous investment in time and money, almost no novel treatments for pain have been developed. There are many factors that contribute to this lack of translation in drug development. The mismatch between the goals of drug development in animals (inhibition of pain-evoked responses) and treatment in humans (restoration of function) is a major problem. To solve this problem, a number of pain-depressed behavioral tests have been developed to assess changes in normal behavior in laboratory animals. The use of home cage wheel running as a pain assessment tool is especially useful in that it is easy to use, provides an objective measurement of the magnitude and duration of pain, and is a clinically relevant method to screen novel drugs. Pain depresses activity in humans and animals, and effective analgesic treatments restore activity. Unlike traditional pain-evoked tests (e.g., hot plate, tail flick, von Frey test), restoration of home cage wheel running evaluates treatments for both antinociceptive efficacy and the absence of disruptive side effects (e.g., sedation, paralysis, nausea). This article reviews the literature using wheel running to assess pain and makes the case for home cage wheel running as an effective and clinically relevant method to screen novel analgesics for therapeutic potential.

Use of home cage wheel running to assess the behavioural effects of administering a mu/delta opioid receptor heterodimer antagonist for spontaneous morphine withdrawal in the rat

Opioid abuse is a major health problem. The objective of the present study was to evaluate the potentially disruptive side effects and therapeutic potential of a novel antagonist (D24M) of the mu-/delta-opioid receptor (MOR/DOR) heterodimer in male rats. Administration of high doses of D24M (1 & 10 nmol) into the lateral ventricle did not disrupt home cage wheel running. Repeated twice daily administration of increasing doses of morphine (5–20 mg/kg) over 5 days depressed wheel running and induced antinociceptive tolerance measured with the hot plate test. Administration of D24M had no effect on morphine tolerance, but tended to prolong morphine antinociception in non-tolerant rats. Spontaneous morphine withdrawal was evident as a decrease in body weight, a reduction in wheel running and an increase in sleep during the normally active dark phase of the circadian cycle, and an increase in wheel running and wakefulness in the normally inactive light phase. Administration of D24M during the dark phase on the third day of withdrawal had no effect on wheel running. These data provide additional evidence for the clinical relevance of home cage wheel running as a method to assess spontaneous opioid withdrawal in rats. These data also demonstrate that blocking the MOR/DOR heterodimer does not produce disruptive side effects or block the antinociceptive effects of morphine. Although administration of D24M had no effect on morphine withdrawal, additional studies are needed to evaluate withdrawal to continuous morphine administration and other opioids in rats with persistent pain.

Analysis of Acute and Chronic Inflammatory Pain‐Depressed Wheel Running in the Rat

Traditional pain‐evoked tests (e.g., hot plate and tail withdrawal tests) do not mimic the disruptive effects of pain on daily life, a primary outcome measure used for pain in humans. This mismatch may contribute to the lack of translation of analgesics from the laboratory to the clinic. To solve this problem, we have previously demonstrated that pain‐depressed home cage wheel running is an objective and clinically relevant measure of the functional consequences of chronic inflammatory pain in the rat (Kandasamy et al., 2016, J. Neurosci. Methods). In this study, we extend this approach by assessing the extent to which acute formalin‐induced inflammatory pain depresses wheel running in the rat. The formalin test is a widely used test of acute nociception where animals are monitored for pain‐like behaviors (i.e., guarding or licking the paw) approximately one hour following formalin injection into the hindpaw. Few studies have examined the functional consequences of acute pain; thus, the aim of this study was to examine the effects of formalin‐induced inflammatory pain on function as measured by home cage wheel running. Rats were housed in individual cages containing a computerized running wheel where they remained for the rest of the experiment. Rats were allowed free access to the wheel 23 hours/day for 7 days prior to the induction of hindpaw inflammation to acquire stable running levels. Wheel running during the 23 hours immediately prior to induction of inflammation was used as the baseline. Inflammation was induced with an intraplantar injection of Complete Freund’s Adjuvant (CFA; 0.1 mL) or formalin (5%, 0.05 mL) into the right hindpaw. As expected, CFA almost completely inhibited wheel running in rats in the 23 hours following administration. Mechanical withdrawal thresholds, as measured by the von Frey test, remained low across the entire testing period. In contrast, the functional effects of a single formalin injection were apparent as indicated by decreased wheel running after the first hour following injection. Wheel running was depressed in the absence of mechanical allodynia, suggesting that inflammatory pain‐induced mechanical allodynia and decreased function are independent of each other. Importantly, these results indicate that continuous monitoring of the functional consequences of pain using home cage wheel running reveal insights into changes in behavior resulting from both acute and chronic pain.Support or Funding InformationSupport for this project was provided by a 2019–20 Faculty Support Grant from the California State University Division of Academic Affairs (RK) and funds from the College of Science at California State University, East Bay (RK).

Voluntary exercise ameliorates anxiogenic effects of acute methamphetamine exposure in Swiss-Webster mice.

The present experiment examined the ability of voluntary exercise (i.e., home-cage wheel running; HCWR) to ameliorate anxiety-like behavior associated with acute methamphetamine exposure in male, Swiss-Webster mice. Mice were permitted access to home-cage running wheels (Exercise), locked home-cage running wheels or no home-cage running wheels (Sedentary) for 6 weeks and then exposed to different methamphetamine doses (vehicle, 0.25, 0.5, or 1.0 mg/kg) once weekly during an 8 h open-field session for 4 weeks. Group differences in hypothalamic-pituitary-adrenal (HPA) activity also were assessed by weighing adrenal glands. It was found that HCWR ameliorated anxiety-like behavior after an injection of either the 0.5 or 1.0 mg/kg methamphetamine dose. Adrenal weights did not differ between Exercise and Sedentary mice. Taken together, these results suggest that voluntary exercise ameliorates the anxiogenic effects of methamphetamine depending on the dose, perhaps via a non-HPA mechanism.

Anti-migraine effect of ∆9-tetrahydrocannabinol in the female rat

Current anti-migraine treatments have limited efficacy and many side effects. Although anecdotal evidence suggests that marijuana is useful for migraine, this hypothesis has not been tested in a controlled experiment. Thus, the present study tested whether administration of ∆9-tetrahydrocannabinol (THC) produces anti-migraine effects in the female rat. Microinjection of the TRPA1 agonist allyl isothiocyanate (AITC) onto the dura mater produced migraine-like pain for 3h as measured by depression of home cage wheel running. Concurrent systemic administration of 0.32 but not 0.1mg/kg of THC prevented AITC-induced depression of wheel running. However, 0.32mg/kg was ineffective when administered 90min after AITC. Administration of a higher dose of THC (1.0mg/kg) depressed wheel running whether rats were injected with AITC or not. Administration of a CB1, but not a CB2, receptor antagonist attenuated the anti-migraine effect of THC. These data suggest that: 1) THC reduces migraine-like pain when administered at the right dose (0.32mg/kg) and time (immediately after AITC); 2) THC's anti-migraine effect is mediated by CB1 receptors; and 3) Wheel running is an effective method to assess migraine treatments because only treatments producing antinociception without disruptive side effects will restore normal activity. These findings support anecdotal evidence for the use of cannabinoids as a treatment for migraine in humans and implicate the CB1 receptor as a therapeutic target for migraine.

Depression of home cage wheel running reveals greater spontaneous morphine withdrawal in rats with persistent pain compared to naïve controls

The widespread use of opioids to treat pain has led to a drastic increase in opioid dependence and overdose fatalities. The transition from the use of opioids to treat pain to abuse is driven in part by withdrawal symptoms associated with the abrupt use of opioids. The objective of the present study is to develop an animal model of withdrawal that mimics the spontaneous withdrawal that occurs in pain patients. We hypothesized that spontaneous opioid withdrawal will cause depression of home cage wheel running. This hypothesis was tested by measuring home cage wheel running before and after subcutaneous implantation of two 75 mg morphine or placebo pellets into male Sprague‐Dawley rats with and without hindpaw inflammation. Administration of the morphine pellets depressed wheel running that recovered to baseline levels by the third day after implantation in naïve rats and by the fifth day after implantation in inflamed rats. Removal of the morphine pellets depressed wheel running for 2 days in naïve animals and over 6 days in rats with an inflamed paw. These data demonstrate that home cage wheel running is a consistent and reliable method to assess spontaneous morphine withdrawal in the rat. Moreover, the magnitude of withdrawal is greatly enhanced in rats with persistent pain.Support or Funding InformationThis study was supported in part by State of Washington Initiative Measure No. 171 (to ATL) and NIH grant NS095097 (to MMM).

Depression of home cage wheel running is an objective measure of spontaneous morphine withdrawal in rats with and without persistent pain

Opioid withdrawal in humans is often subtle and almost always spontaneous. In contrast, most preclinical studies precipitate withdrawal by administration of an opioid receptor antagonist such as naloxone. These animal studies rely on measurement of physiological symptoms (e.g., wet dog shakes) in the period immediately following naloxone administration. To more closely model the human condition, we tested the hypothesis that depression of home cage wheel running will provide an objective method to measure the magnitude and duration of spontaneous morphine withdrawal. Rats were allowed access to a running wheel in their home cage for 8days prior to implantation of two 75mg morphine or placebo pellets. The pellets were removed 3 or 5days later to induce spontaneous withdrawal. In normal pain-free rats, removal of the morphine pellets depressed wheel running for 48h compared to rats that had placebo pellets removed. Morphine withdrawal-induced depression of wheel running was greatly enhanced in rats with persistent inflammatory pain induced by injection of Complete Freund's Adjuvant (CFA) into the hindpaw. Removal of the morphine pellets following 3days of treatment depressed wheel running in these rats for over 6days. These data demonstrate that home cage wheel running provides an objective and more clinically relevant method to assess spontaneous morphine withdrawal compared to precipitated withdrawal in laboratory rats. Moreover, the enhanced withdrawal in rats with persistent inflammatory pain suggests that pain patients may be especially susceptible to opioid withdrawal.

Depression of home cage wheel running: a reliable and clinically relevant method to assess migraine pain in rats

BackgroundThe development of new anti-migraine treatments is limited by the difficulty inassessing migraine pain in laboratory animals. Depression of activity is one of the few diagnostic criteria formigraine that can be mimicked in rats. The goal of the present study was to test the hypothesis thatdepression of home cage wheel running is a reliable and clinically relevant method to assess migraine painin rats.MethodsAdult female rats were implanted with a cannula to inject allyl isothiocyanate (AITC) onto the dura to induce migraine pain, as has been shown before. Rats recovered from implantation surgery for 8 days in cages containing a running wheel. Home cage wheel running was recorded 23 h a day. AITC and the migraine medication sumatriptan were administered in the hour prior to onset of the dark phase.ResultsAdministration of AITC caused a concentration-dependent decrease in wheel running that lasted 3 h. The duration and magnitude of AITC-induced depression of wheel running was consistent following three repeated injections spaced 48 h apart. Administration of sumatriptan attenuated AITC-induced depressionof wheel running when a large dose (1 mg/kg) was administered immediately following AITC administration. Wheel running patterns did not change when sumatriptan was given to naïve rats.ConclusionsThese data indicate that home cage wheel running is a sensitive, reliable, and clinically relevant method to assess migraine pain in the rat.

Analysis of inflammation-induced depression of home cage wheel running in rats reveals the difference between opioid antinociception and restoration of function

Opioids are effective at inhibiting responses to noxious stimuli in rodents, but have limited efficacy and many side effects in chronic pain patients. One reason for this disconnect is that nociception is typically assessed using withdrawal from noxious stimuli in animals, whereas chronic pain patients suffer from abnormal pain that disrupts normal activity. We hypothesized that assessment of home cage wheel running in rats would provide a much more clinically relevant method to assess opioid efficacy to restore normal behavior. Intraplantar injection of Complete Freund’s Adjuvant (CFA) into the right hindpaw depressed wheel running and caused mechanical allodynia measured with the von Frey test in both male and female rats. Administration of an ED50 dose of morphine (3.2mg/kg) reversed mechanical allodynia, but did not reverse CFA-induced depression of wheel running. In contrast, administration of a low dose of morphine (1.0mg/kg) restored running for one hour in both sexes, but had no effect on mechanical allodynia. Administration of the atypical opioid buprenorphine had no effect on inflammation-induced depression of wheel running in male or female rats, but attenuated mechanical allodynia in male rats. Administration of buprenorphine and higher doses of morphine depressed wheel running in non-inflamed rats, suggesting that the side effects of opioids interfere with restoration of function. These data indicate that restoration of pain-depressed function requires antinociception in the absence of disruptive side effects. The disruptive side effects of opioids are consistent with the major limitation of opioid use in human pain patients.

Depression of Home Cage Wheel Running: A Novel Method to Assess Spontaneous Migraine Pain

Migraine is the most common neurological disorder in the world, yet current treatments have limited efficacy and repeated administration can cause medication‐overuse headache. Although new treatments are desperately needed, the difficulty of assessing migraine pain in laboratory animals has limited drug development. Most animal studies of migraine assess mechanical allodynia in the face and hindpaw of the rodent. Although useful, this measure does not assess spontaneous, unevoked headache pain – a key characteristic of migraine. The objective of the present study is to test the hypothesis that home cage wheel running is a sensitive and objective method to assess spontaneous migraine pain in the rat – an approach that mimics the decrease in activity used to diagnose migraine in humans. Adult male and female rats were implanted with a cranial cannula aimed at the periosteal space above the dura mater. Rats were allowed to recover for 8 days in their home cage with unlimited access to a running wheel. Migraine pain was induced by microinjecting 10μL of 10% allyl isothiocyanate (AITC) onto the dura. Control rats were injected with mineral oil. Rats were injected with the anti‐migraine agent sumatriptan (1 mg/kg, s.c.) or saline 30 min following the intracranial injection. Administration of AITC into the periosteal space caused a decrease in running in both male and female rats that lasted 4–7 hrs. Administration of sumatriptan prevented this depression of wheel running. The fact that sumatriptan prevented migraine pain‐induced depression of home cage wheel‐running demonstrates that home cage wheel running is a sensitive and reliable measure of spontaneous headache pain in rats.Support or Funding InformationThe study was supported by State of Washington Initiative Measure No. 171 and Washington State University.

Home cage wheel running is an objective and clinically relevant method to assess inflammatory pain in male and female rats

BackgroundThe assessment of nociception in preclinical studies is undergoing a transformation from pain-evoked to pain-depressed tests to more closely mimic the effects of clinical pain. Many inflammatory pain-depressed behaviors (reward seeking, locomotion) have been examined, but these tests are limited because of confounds such as stress and difficulties in quantifying behavior. New methodThe present study evaluates home cage wheel running as an objective method to assess the magnitude and duration of inflammatory pain in male and female rats. ResultsInjection of Complete Freund's Adjuvant (CFA) into the right hindpaw to induce inflammatory pain almost completely inhibited wheel running for 2 days in male and female rats. Wheel running gradually returned to baseline levels within 12 days despite persistent mechanical hypersensitivity (von Frey test). Comparison with existing methodsContinuously monitoring home cage wheel running improves on previous studies examining inflammatory pain-depressed wheel running because it is more sensitive to noxious stimuli, avoids the stress of removing the rat from its cage for testing, and provides a complete analysis of the time course for changes in nociception. ConclusionsThe present data indicate that home cage wheel running is a clinically relevant method to assess inflammatory pain in the rat. The decrease in activity caused by inflammatory pain and subsequent gradual recovery mimics the changes in activity caused by pain in humans. The tendency for pain-depressed wheel running to be greater in female than male rats is consistent with the tendency for women to be at greater risk of chronic pain than men.

Low amplitude entrainment of mice and the impact of circadian phase on behavior tests

A tremendous increase in the use of genetically engineered mice as experimental animals has led to increased scrutiny of mouse models generally and mouse behavioral paradigms specifically. Although mice are nocturnal, for practical reasons, most experimental procedures, including behavioral studies, are conducted during their inactive, sleep phase. Accumulating evidence indicates that myriad behavioral, cellular and biochemical processes fluctuate with circadian rhythmicity; however, time of day at which experiments are conducted is rarely controlled. The impact of circadian phase on the reliability of experimental results has received little attention and the present data are conflicting. This study addressed two questions. First, will laboratory mice in a typical animal care facility entrain to a low amplitude light cycle using bright/dim rather than light/dark cycles? A positive answer will make reversing photocycle easy to implement in any facility as dim light suitable for animal husbandry and behavioral testing can substitute for darkness during work hours. By monitoring home cage wheel running, we examined the effectiveness of a dim/bright photocycle as a zeitgeiber. We found that mice subjected to dim/bright photocycles effectively entrained such that their subjective night and activity onset coincided with the beginning of the dim light period, suggesting a potential strategy for standardization and management of circadian phase in nocturnal animals. In a second experiment, we asked what effect circadian phase has on behavioral performance in commonly used mouse behavioral tests. We found no main effect of circadian phase on outcome in open field activity, elevated plus maze emotionality, water maze spatial memory, novel object exploration and hyperactivity in response to amphetamine; however, we observed occasional interactions between circadian phase and both strain and sex that were neither consistent nor systematic. These data suggest that the tests examined here are relatively impervious to circadian phase. In general, testing mice during their active phase is more suitable for behavioral studies; a reversed dim/bright photocycle potentially offers one practical strategy for managing rodents' circadian cycles.

Presynaptic inhibition involved in periaqueductal gray suppression of the activities of dorsal horn neurons

Opioids are effective at inhibiting responses to noxious stimuli in rodents, but have limited efficacy and many side effects in chronic pain patients. One reason for this disconnect is that nociception is typically assessed using withdrawal from noxious stimuli in animals, whereas chronic pain patients suffer from abnormal pain that disrupts normal activity. We hypothesized that assessment of home cage wheel running in rats would provide a much more clinically relevant method to assess opioid efficacy to restore normal behavior. Intraplantar injection of Complete Freund’s Adjuvant (CFA) into the right hindpaw depressed wheel running and caused mechanical allodynia measured with the von Frey test in both male and female rats. Administration of an ED50 dose of morphine (3.2 mg/kg) reversed mechanical allodynia, but did not reverse CFA-induced depression of wheel running. In contrast, administration of a low dose of morphine (1.0 mg/kg) restored running for one hour in both sexes, but had no effect on mechanical allodynia. Administration of the atypical opioid buprenorphine had no effect on inflammation-induced depression of wheel running in male or female rats, but attenuated mechanical allodynia in male rats. Administration of buprenorphine and higher doses of morphine depressed wheel running in non-inflamed rats, suggesting that the side effects of opioids interfere with restoration of function. These data indicate that restoration of pain-depressed function requires antinociception in the absence of disruptive side effects. The disruptive side effects of opioids are consistent with the major limitation of opioid use in human pain patients.