Review
Comparison of operant escape and reflex tests of nociceptive sensitivity

https://doi.org/10.1016/j.neubiorev.2015.01.022Get rights and content

Highlights

  • Stimulus–response functions for reflexes and operant escape are compared.

  • Assessment of neuropathic pain is compared for reflexes and operant escape.

  • Sex, age and stress effects on reflex and escape responding are compared.

  • Reflex and escape effects of morphine and cholinergic denervation are compared.

  • Control procedures for reflex and escape effects are discussed.

Abstract

Testing of reflexes such as flexion/withdrawal or licking/guarding is well established as the standard for evaluating nociceptive sensitivity and its modulation in preclinical investigations of laboratory animals. Concerns about this approach have been dismissed for practical reasons – reflex testing requires no training of the animals; it is simple to instrument; and responses are characterized by observers as latencies or thresholds for evocation. In order to evaluate this method, the present review summarizes a series of experiments in which reflex and operant escape responding are compared in normal animals and following surgical models of neuropathic pain or pharmacological intervention for pain. Particular attention is paid to relationships between reflex and escape responding and information on the pain sensitivity of normal human subjects or patients with pain. Numerous disparities between results for reflex and operant escape measures are described, but the results of operant testing are consistent with evidence from humans. Objective reasons are given for experimenters to choose between these and other methods of evaluating the nociceptive sensitivity of laboratory animals.

Section snippets

Comparison of reflex and operant escape responses to nociceptive stimulation of normal monkeys, rats and humans

For stimuli of sufficient intensity to elicit reflexes, response latencies distinguish flexion/withdrawal reflexes from operant escape responses. In studies of Macaca speciosa monkeys and Long-Evans rats, electrical stimulation has been delivered to one hindlimb while simultaneously recording flexion/withdrawal of the stimulated limb and operant escape. Manual lever pulls (monkeys) or bar press responses (rats) with either hand terminate stimulation of the leg that responds with

CNS circuits in support of reflex and operant escape responding

Separate neural circuits participate in the generation of reflex and operant escape responses. Spinal neurons provide output to the periphery or the brain stem or the cerebrum for the 3 behavioral categories emphasized in this review.

Laboratory animal models of neuropathic pain following peripheral injury

Different methods of ligating or partially transecting a peripheral nerve have been utilized to provide animal models of neuropathic pain. In these models, flexion/withdrawal reflexes are enhanced (shorter latencies for thermal stimulation or lower thresholds for mechanical stimulation) for the treated hindlimb relative to the contralateral hindlimb when similarly stimulated. One of these models, chronic constriction injury (CCI) of the sciatic nerve, produces an ipsilateral enhancement of

Effects of spinal cord injury (SCI) on nociceptive reflexes and on pain sensitivity

Operant escape testing has consistently confirmed clinical evidence from humans (King, 1957, White and Sweet, 1969) that interruption of the lateral spinothalamic tract initially produces contralateral hypoalgesia, below the level of the lesion (Vierck et al., 1971, Vierck et al., 1983b, Vierck et al., 1986, Vierck et al., 1990, Vierck et al., 1995, Vierck et al., 2000, Vierck and Light, 1999, Vierck and Light, 2000, Vierck and Light, 2002, Vierck and Luck, 1979). In contrast to the

Stress-induced analgesia or hyperalgesia?

An extensive literature involving laboratory animal subjects and reflex measures has portrayed acute psychological stress as analgesic (Bodnar et al., 1980). Probably, stress-induced hypoalgesia is a real but time-limited effect, judging from anecdotal reports of performance feats during stress associated with painful injury. However, it is problematic to evaluate pain sensitivity during presentation of most stressors, because of distraction. Therefore, the standard experimental paradigm for

Sex differences in reflex and operant escape responding to thermal stimulation

Whether there are meaningful differences in pain sensitivity for males and females has received considerable attention in recent years. Human psychophysical studies have concluded that females are more sensitive to pain (Riley et al., 1998), and there is a higher prevalence of chronic pain conditions among females (Unruh, 1996). Interpretation of these studies is complicated by psychosocial influences that can predispose females to exaggerate pain more than males (Jackson et al., 2005, Kallai

Effect of age on escape and thermal preference testing

Escape testing of female Fischer 344 × Brown Norway rats (bred for longevity) has revealed hyperalgesia for 15 °C, 10 °C, 42 °C, and 44.5 °C stimulation of 32 month-old females, compared to 8 or 16 month-old females (Yezierski et al., 2010). Cold sensitivity increases with age to a greater extent than heat sensitivity. This has been confirmed by changing our escape task from an opposition of bright light vs. heat or cold to an opposition of heat vs. cold stimulation. Thermal preference testing of

Effects of systemic morphine on pain sensitivity of humans, monkeys and rats

Systemic μ-opioid agonists have long been considered the most powerful pharmacological agents for inhibition of pain. However, attempts to understand mechanisms of opioid hypoalgesia have been misdirected by the prevalent methods of antinoceptive testing of humans and laboratory animals. Primary considerations are that: (A) reflexes do not reveal opioid actions at sites throughout the neuraxis known to contain opioid receptors (Becerra et al., 2013, Hagelberg et al., 2012, Maarrawi et al., 2007

Loss of cholinergic basal forebrain neurons and systemic cholinergic antagonism

Cholinergic agonists are antinociceptive when administered intrathecally to humans (Bartolini et al., 2011, Jones and Dunlop, 2007), and nociceptive reflexes are attenuated following intrathecal application of cholinergic agonists (Bouaziz et al., 1995, Eisenach and Gebhart, 1995, Gillberg et al., 1990, Hartvig et al., 1989, Yaksh et al., 1985). Also, reflex responses are inhibited by systemic or intracerebral administration of cholinergic agonists or acetylcholinesterase antagonists (Abelson

Controls and other methodological considerations

It is obvious that controls are needed to insure that effects on behavioral reactions to stimulation represent a modulation of nociceptive sensitivity. More accurately, the importance of controls is understood for operant escape, but this has not been the case for reflex tests. For example, morphine is expected to inhibit pain selectively. However, doses of morphine too low to attenuate reflexes can reduce escape responding to non-nociceptive stimulation. Similar procedures could have

Semantics: pain modulation vs. motor control

Different categories of altered pain sensitivity, such as allodynia, hyperalgesia, hypoalgesia or analgesia have been defined relative to thresholds for consciously perceived pain intensity (IASP, 1994). Clearly, these definitions do not relate or refer to any characteristic of reflex responses to nociceptive stimulation. And yet, reports of laboratory animal experiments have consistently referred to effects of experimental manipulations on nociceptive reflexes as allodynia, hyperalgesia, or

Pain research strategies

Preclinical testing of pharmacological agents with a potential for pain modulation has almost exclusively utilized reflex tests. Much of this screening is conducted by pharmaceutical companies without publication of the drugs tested or the results. The strategy appears to be that reflexes provide an efficient means of screening the actions of multiple drugs in laboratory animals, and then promising agents can be evaluated with conscious reports by humans. This is a flawed approach on numerous

Summary and implications

The investigation of unmotivated, innate reflexes that primarily serve motor control from the brain stem is not relevant to pain. Study of a simple circuit from the periphery to spinal interneurons and then to motoneurons will not reveal functions of different and more extensive circuits from the periphery to the cerebral cortex. Activation of the cerebral targets of nociceptive projection is necessary for evocation of pain sensations and for motivated programming of escape responses (Bushnell

Conflict of interest

One of the authors (C.J.V.) is a board member of Neuroanalytics, a company that has developed automated equipment for human psychophysical testing and for operant and reflex testing of laboratory animals.

Acknowledgements

Research summarized in this review was supported by: NIH grant NS-07261, funds supplied by the University of Florida Department of Neuroscience and Brain Institute, NIA grant RAG-031821, and a grant for international collaboration from the International Association for the Study of Pain, Scan Design and the Bruun Foundation. The technical support of Jean Kaufman and Karen Murphy is gratefully acknowledged, for their assistance in the studies reported here, involving expert handling and care of

References (265)

  • J.D. Brioni et al.

    Nicotinic receptor agonists exhibit anxiolytic-like effects on the elevated plus-maze test

    Eur. J. Pharmacol.

    (1993)
  • R. Bronsing et al.

    Modulation of cutaneous reflexes in hindlimb muscles during locomotion in the freely walking rat: a model for studying cerebellar involvement in the adaptive control of reflexes during rhythmic movements

    Prog. Brain Res.

    (2005)
  • B. Bussel et al.

    Late flexion reflex in paraplegic patients. Evidence for a spinal stepping generator

    Brain Res. Bull.

    (1989)
  • P. Carrive et al.

    A reassessment of stress-induced analgesia in the rat using an unbiased method

    Pain

    (2011)
  • R.C. Castillo et al.

    Longitudinal relationships between anxiety, depression, and pain: results from a two-year cohort study of lower extremity trauma patients

    Pain

    (2013)
  • C.W. Chan et al.

    Subjective pain sensation is linearly correlated with the flexion reflex in man

    Brain Res.

    (1989)
  • M.D. Christensen et al.

    Mechanical and thermal allodynia in chronic central pain following spinal cord injury

    Pain

    (1996)
  • F.C. Colpaert et al.

    Opiate self-administration as a measure of chronic nociceptive pain in arthritic rats

    Pain

    (2001)
  • B.Y. Cooper et al.

    Measurement of pain and morphine hyperalgesia in monkeys

    Pain

    (1986)
  • B.Y. Cooper et al.

    Selective reduction of second pain sensations by systemic morphine in humans

    Pain

    (1986)
  • K.D. Craig et al.

    Genuine, suppressed and faked facial behavior during exacerbation of chronic low back pain

    Pain

    (1991)
  • K.D. Davis et al.

    Topical application of clonidine relieves hyperalgesia in patients with sympathetically maintained pain

    Pain

    (1991)
  • R. Dowman

    Spinal and supraspinal correlates of nociception in man

    Pain

    (1991)
  • G.M. Drew et al.

    Responses of spinal neurones to cutaneous and dorsal root stimuli in rats with mechanical allodynia after contusive spinal cord injury

    Brain Res.

    (2001)
  • G.M. Drew et al.

    Mechanical allodynia following contusion injury of the rat spinal cord is associated with loss of GABAergic inhibition in the dorsal horn

    Pain

    (2004)
  • M. Dufosse et al.

    The postural reaction to the drop of a hindlimb support in the standing cat remains following sensorimotor cortical ablation

    Neurosci. Lett.

    (1985)
  • S.E. File et al.

    The role of the dorsal hippocampal serotonergic and cholinergic systems in the modulation of anxiety

    Pharmacol. Biochem. Behav.

    (2000)
  • Y. Finkelstein et al.

    Dynamics of cholinergic synaptic mechanisms in rat hippocampus after stress

    Brain Res.

    (1985)
  • D.P. Finn et al.

    Behavioral, central monoaminergic and hypothalamo–pituitary–adrenal axis correlates of fear-conditioned analgesia in rats

    Neuroscience

    (2006)
  • N.B. Finnerup et al.

    Phenotypes and predictors of pain following traumatic spinal cord injury: a prospective study

    J. Pain

    (2014)
  • G.M. Gilad et al.

    Stress-induced activation of the hippocampal cholinergic system and the pituitary–adrenocortical axis

    Brain Res.

    (1985)
  • R.F. Gledhill et al.

    Demyelination and remyelination after acute spinal cord compression

    Exp. Neurol.

    (1973)
  • C. Gouarderes et al.

    High resolution radioautographic localization of [125I]FK-33-824-labelled mu opioid receptors in the spinal cord of normal and deafferented rats

    Neuroscience

    (1991)
  • A.J. Gower

    Effects of acetylcholine agonists and antagonists on yawning and analgesia in the rat

    Eur. J. Pharmacol.

    (1987)
  • R.H. Gracely et al.

    Ratio scales of sensory and affective verbal pain descriptors

    Pain

    (1978)
  • S.M. Gustin et al.

    Thalamic activity and biochemical changes in individuals with neuropathic pain after spinal cord injury

    Pain

    (2014)
  • N. Hagelberg et al.

    Striatal mu-opioid receptor availability predicts cold pressor pain threshold in healthy human subjects

    Neurosci. Lett.

    (2012)
  • M.A. Hagenaars et al.

    Updating freeze: aligning animal and human research

    Neurosci. Biobehav. Rev.

    (2014)
  • J.X. Hao et al.

    Photochemically induced transient spinal ischemia induces behavioral hypersensitivity to mechanical and cold stimuli, but not to noxious-heat stimuli, in the rat

    Exp. Neurol.

    (1992)
  • K. Hargreaves et al.

    A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia

    Pain

    (1988)
  • S. Haroutiunian et al.

    The neuropathic component in persistent postsurgical pain: a systematic literature review

    Pain

    (2013)
  • J.L. Harrison et al.

    Cold-evoked pain varies with skin type and cooling rate: a psychophysical study in humans

    Pain

    (1999)
  • N. Hayashi et al.

    Vascular responses to fear-induced stress in humans

    Physiol. Behav.

    (2009)
  • K. Hole et al.

    The tail-flick and formalin tests in rodents: changes in skin temperature as a confounding factor

    Pain

    (1993)
  • C.E. Hulsebosch et al.

    Mechanisms of chronic central neuropathic pain after spinal cord injury

    Brain Res. Rev.

    (2009)
  • K.S. Abelson et al.

    Intravenously administered oxotremorine and atropine, in doses known to affect pain threshold, affect the intraspinal release of acetylcholine in rats

    Pharmacol. Toxicol.

    (2002)
  • N. Andrews et al.

    Spontaneous burrowing behaviour in the rat is reduced by peripheral nerve injury or inflammation associated pain

    Eur. J. Pain

    (2012)
  • A.A. Argyriou et al.

    Incidence and characteristics of peripheral neuropathy during oxaliplatin-based chemotherapy for metastatic colon cancer

    Acta Oncol.

    (2007)
  • C. Aveline et al.

    Pain and recovery after total knee arthroplasty: a twelve months follow-up after a prospective randomized study evaluating nefopam and ketamine for early rehabilitation

    Clin. J. Pain.

    (2013)
  • A. Bartolini et al.

    Analgesic and antineuropathic drugs acting through central cholinergic mechanisms

    Recent Pat. CNS Drug Discov.

    (2011)
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