Plasma corticosterone levels in mouse models of pain

Background: Pain markedly activates the hypothalamic-pituitary-adrenal (HPA) axis and increases plasma corticosterone release interfering significantly with nociceptive behaviour as well as the mechanism of action of analgesic drugs. Aims/Methods: In the present study, we monitored the time course of circulating corticosterone in two mouse strains (C57Bl/6 and Balb/C) under different pain models. In addition, the stress response was investigated following animal handling, intrathecal (i.t.) manipulation and habituation to environmental conditions commonly used in nociceptive experimental assays. We also examined the influence of within-cage order of testing on plasma corticosterone. Results: Subcutaneous injection of capsaicin precipitated a prompt stress response whereas carrageenan and complete Freund's adjuvant induced an increased corticosterone release around the third hour post-injection. However, carrageenan induced a longer increased corticosterone in C57Bl/6 mice. In partial sciatic nerve ligation, neuropathic pain model corticosterone increased only in the first days whereas mechanical hypersensitivity remained much longer. Animal handling also represents an important stressor whereas the i.t. injection per se does not exacerbate the handling-induced stress response. Moreover, the order of testing animals from the same cage does not interfere with plasma corticosterone levels in the intrathecal procedure. Animal habituation to the testing apparatus also does not reduce the immediate corticosterone increase as compared with non-habituated mice. Conclusion: Our data indicate that HPA axis activation in acute and chronic pain models is time dependent and may be dissociated from evoked hyperalgesia. Therefore, HPA-axis activation represents an important variable to be considered when designing experimental assays of persistent pain as well as for interpretation of data.

Local pain and spreading hyperalgesia induced by intramuscular injection of nerve growth factor are not reduced by local anesthesia of the muscle

Injections with local anesthesia for therapeutic and diagnostic purposes are common clinical practice. This double-blind placebo controlled study explores the rational of local anesthetic blocks for the detection of muscle pain as the primary generator in spreading hyperalgesic conditions.

Influence of local anaesthesia on pain and distress induced by two bloodless castration methods in young lambs

To assess short- and long-term effects of bloodless castration methods with and without local anaesthesia, behavioural and cortisol responses of lambs were used as indicators of pain and distress. Seventy lambs, aged 2-7 days, were control-handled or castrated by Burdizzo or rubber ring methods with and without local anaesthesia. Either 5 mL of diluted lidocaine (4 mg/kg) or physiological sodium chloride solution was distributed in both spermatic cords and the scrotal neck. The serum cortisol response was monitored for 48 h, and behavioural and clinical traits were followed for three months. Local anaesthesia tended to reduce behavioural and cortisol responses after Burdizzo castration and provided a significant reduction after rubber ring castration. Prolonged pain after rubber ring castration with anaesthesia was not evident. If combined with local anaesthesia, both the rubber ring and the Burdizzo methods are acceptable methods for castration of lambs up to one week of age.

[Influence of local anaesthesia on pain and distress induced by bloodless castration methods in young lambs]

Behavioural and cortisol responses of lambs were used as indicators of pain and distress to assess short- and long-term effects of bloodless castration methods with and without local anaesthesia. Eighty lambs, aged 2-7 days, were control handled or castrated by crushing- Burdizzo- or rubber ring method with and without local anaesthesia. Either 4 mg/kg diluted lidocaine, or corresponding volumes of physiologic sodium chloride solution were distributed in both spermatic cords and the scrotal neck. The serum cortisol response was monitored for 48 h, behavioural- and clinical traits over a 3-month period. The crushing castration method was excluded from the study after 10 lambs had been castrated, since this method showed severe local reactions. Local anaesthesia significantly reduced behavioural and cortisol responses after rubber ring castration and tendentially after Burdizzo castration. Prolonged or chronic pain after rubber ring castration with anaesthesia was not evident. If combined with local anaesthesia, both the rubber ring and the Burdizzo method are acceptable methods for castration of lambs up to one week of age.

[Influence of local anaesthesia on short- and long-term pain induced in calves by three bloodless castration methods]

Behavioural and cortisol responses of calves were used as indicators of pain to assess short- and long-term effects of three bloodless castration methods with and without local anaesthesia. Eighty calves, aged 21 to 28 days, were control handled (20) or castrated by Burdizzo (25), rubber ring (25), or crushing technique (10). Either a total volume of 10 ml of Lidocaine or NaCl was distributed in both spermatic cords and the scrotal neck. The plasma cortisol response was monitored for 72 hours, and behavioural and clinical traits over a three-month period. Castration success was assessed by degree of atrophy and histological tissue examination. The crushing technique cannot be recommended due to incomplete castration success, and the evaluation was stopped after 10 animals. Local anaesthesia reduced the level of indicators of acute pain after Burdizzo and rubber ring technique. It did, however, not result in a totally painless castration. When castration is performed at the age of 3 to 4 weeks, the rubber ring but not the Burdizzo method showed evidence of chronic pain lasting for several weeks.

Effect of local anaesthesia on short- and long-term pain induced by two bloodless castration methods in calves

Behavioural and cortisol responses of calves were used as indicators of pain to assess short- and long-term effects of bloodless castration methods with and without local anaesthesia. Seventy calves, aged 21-28 days, were control handled (20) or castrated using the Burdizzo (25) or rubber ring technique (25). Either 10 mL lidocaine or NaCl were distributed in both spermatic cords and the scrotal neck. The plasma cortisol response was recorded for 72 h, and behavioural and clinical traits monitored over a three month period. Local anaesthesia reduced the level of indicators of acute pain after both the Burdizzo and rubber ring techniques. It did not, however, result in a totally painless castration. As there was evidence of chronic pain lasting for several weeks after rubber ring castration, the Burdizzo method is judged to be preferable to the rubber ring technique.

Dysregulation of voltage-gated sodium channels by ubiquitin ligase NEDD4-2 in neuropathic pain

Peripheral neuropathic pain is a disabling condition resulting from nerve injury. It is characterized by the dysregulation of voltage-gated sodium channels (Navs) expressed in dorsal root ganglion (DRG) sensory neurons. The mechanisms underlying the altered expression of Na(v)s remain unknown. This study investigated the role of the E3 ubiquitin ligase NEDD4-2, which is known to ubiquitylate Navs, in the pathogenesis of neuropathic pain in mice. The spared nerve injury (SNI) model of traumatic nerve injury-induced neuropathic pain was used, and an Na(v)1.7-specific inhibitor, ProTxII, allowed the isolation of Na(v)1.7-mediated currents. SNI decreased NEDD4-2 expression in DRG cells and increased the amplitude of Na(v)1.7 and Na(v)1.8 currents. The redistribution of Na(v)1.7 channels toward peripheral axons was also observed. Similar changes were observed in the nociceptive DRG neurons of Nedd4L knockout mice (SNS-Nedd4L(-/-)). SNS-Nedd4L(-/-) mice exhibited thermal hypersensitivity and an enhanced second pain phase after formalin injection. Restoration of NEDD4-2 expression in DRG neurons using recombinant adenoassociated virus (rAAV2/6) not only reduced Na(v)1.7 and Na(v)1.8 current amplitudes, but also alleviated SNI-induced mechanical allodynia. These findings demonstrate that NEDD4-2 is a potent posttranslational regulator of Na(v)s and that downregulation of NEDD4-2 leads to the hyperexcitability of DRG neurons and contributes to the genesis of pathological pain.

Nerve Injury-Induced Neuropathic Pain Causes Disinhibition of the Anterior Cingulate Cortex

Neuropathic pain caused by peripheral nerve injury is a debilitating neurological condition of high clinical relevance. On the cellular level, the elevated pain sensitivity is induced by plasticity of neuronal function along the pain pathway. Changes in cortical areas involved in pain processing contribute to the development of neuropathic pain. Yet, it remains elusive which plasticity mechanisms occur in cortical circuits. We investigated the properties of neural networks in the anterior cingulate cortex (ACC), a brain region mediating affective responses to noxious stimuli. We performed multiple whole-cell recordings from neurons in layer 5 (L5) of the ACC of adult mice after chronic constriction injury of the sciatic nerve of the left hindpaw and observed a striking loss of connections between excitatory and inhibitory neurons in both directions. In contrast, no significant changes in synaptic efficacy in the remaining connected pairs were found. These changes were reflected on the network level by a decrease in the mEPSC and mIPSC frequency. Additionally, nerve injury resulted in a potentiation of the intrinsic excitability of pyramidal neurons, whereas the cellular properties of interneurons were unchanged. Our set of experimental parameters allowed constructing a neuronal network model of L5 in the ACC, revealing that the modification of inhibitory connectivity had the most profound effect on increased network activity. Thus, our combined experimental and modeling approach suggests that cortical disinhibition is a fundamental pathological modification associated with peripheral nerve damage. These changes at the cortical network level might therefore contribute to the neuropathic pain condition.

Acupuncture-induced changes of pressure pain threshold are mediated by segmental inhibition-a randomized controlled trial

Our aim was to distinguish between spinal and supraspinal mechanisms in the intact nervous system by comparing homosegmental and heterosegmental effects of electroacupuncture (EA) and manual acupuncture (MA) on sensory perception in healthy volunteers by means of quantitative sensory testing. Seventy-two healthy volunteers were randomly assigned to receive either MA or EA at SP 6, SP 9, GB 39, and ST 36 at the left leg or relaxed for 30 minutes (control group [CG]). Blinded examiners assessed 13 sensory modalities (thermal and mechanical detection and pain thresholds) at the upper arms and lower legs before and after intervention by means of a standardized quantitative sensory testing battery. Change scores of all 13 sensory thresholds were compared between groups. The main outcome measure was the change score of the pressure pain threshold (PPT). There were no baseline differences between groups. Pressure pain threshold change scores at the lower left leg, in the same segment as the needling site, differed significantly (P = 0.008) between the EA (median: 103.01 kPa) and CG groups (median: 0.00 kPa) but not between the MA (median: 0.00 kPa) and CG groups. No further significant change score differences were found between one of the acupuncture groups and the CG. The PPT can be changed by EA. The PPT increase was confined to the segment of needling, which indicates that it is mainly mediated by segmental inhibition in the spinal cord. This underscores the importance of segmental needling and electrical stimulation in clinical practice.

Agmatine reverses pain induced by inflammation, neuropathy, and spinal cord injury

Antagonists of glutamate receptors of the N-methyl-d-aspartate subclass (NMDAR) or inhibitors of nitric oxide synthase (NOS) prevent nervous system plasticity. Inflammatory and neuropathic pain rely on plasticity, presenting a clinical opportunity for the use of NMDAR antagonists and NOS inhibitors in chronic pain. Agmatine (AG), an endogenous neuromodulator present in brain and spinal cord, has both NMDAR antagonist and NOS inhibitor activities. We report here that AG, exogenously administered to rodents, decreased hyperalgesia accompanying inflammation, normalized the mechanical hypersensitivity (allodynia/hyperalgesia) produced by chemical or mechanical nerve injury, and reduced autotomy-like behavior and lesion size after excitotoxic spinal cord injury. AG produced these effects in the absence of antinociceptive effects in acute pain tests. Endogenous AG also was detected in rodent lumbosacral spinal cord in concentrations similar to those previously detected in brain. The evidence suggests a unique antiplasticity and neuroprotective role for AG in processes underlying persistent pain and neuronal injury.

Forebrain mechanisms of nociception and pain: Analysis through imaging

Pain is a unified experience composed of interacting discriminative, affective-motivational, and cognitive components, each of which is mediated and modulated through forebrain mechanisms acting at spinal, brainstem, and cerebral levels. The size of the human forebrain in relation to the spinal cord gives anatomical emphasis to forebrain control over nociceptive processing. Human forebrain pathology can cause pain without the activation of nociceptors. Functional imaging of the normal human brain with positron emission tomography (PET) shows synaptically induced increases in regional cerebral blood flow (rCBF) in several regions specifically during pain. We have examined the variables of gender, type of noxious stimulus, and the origin of nociceptive input as potential determinants of the pattern and intensity of rCBF responses. The structures most consistently activated across genders and during contact heat pain, cold pain, cutaneous laser pain or intramuscular pain were the contralateral insula and anterior cingulate cortex, the bilateral thalamus and premotor cortex, and the cerebellar vermis. These regions are commonly activated in PET studies of pain conducted by other investigators, and the intensity of the brain rCBF response correlates parametrically with perceived pain intensity. To complement the human studies, we developed an animal model for investigating stimulus-induced rCBF responses in the rat. In accord with behavioral measures and the results of human PET, there is a progressive and selective activation of somatosensory and limbic system structures in the brain and brainstem following the subcutaneous injection of formalin. The animal model and human PET studies should be mutually reinforcing and thus facilitate progress in understanding forebrain mechanisms of normal and pathological pain.

The impact of neurodynamic testing on the perception of experimentally induced muscle pain

Neurodynamic tests such as the straight leg raising (SLR) and slump test are frequently used for assessment of mechanosensitivity of neural tissues. However, there is ongoing debate in the literature regarding the contributions of neural and non-neural tissues to the elicited symptoms because many structures are affected by these tests. Sensitizing manoeuvres are limb or spinal movements added to neurodynamic tests, which aim to identify the origin of the symptoms by preferentially loading or unloading neural structures. A prerequisite for the use of sensitizing manoeuvres to identify neural involvement is that the addition of sensitizing manoeuvres has no impact on pain perception when the origin of the pain is non-neural. In this study, experimental muscle pain was induced by injection of hypertonic saline in tibialis anterior or soleus in 25 asymptomatic, naive volunteers. A first experiment investigated the impact of hip adduction, abduction, medial and lateral rotation in the SLR position. In a second experiment, the different stages of the slump test were examined. The intensity and area of experimentally induced muscle pain did not increase when sensitizing manoeuvres were added to the SLR or throughout the successive stages of the slump test. The findings of this study lend support to the validity of the use of sensitizing manoeuvres during neurodynamic testing. (C) 2004 Elsevier Ltd. All rights reserved.

Reduced variability of postural strategy prevents normalization of motor changes induced by back pain: A risk factor for chronic trouble?

Variability is fundamental to biological systems and is important in posturomotor learning and control. Pain induces a protective postural strategy, although variability is normally preserved. If variability is lost, does the normal postural strategy return when pain stops? Sixteen subjects performed arm movements during control trials, when the movement evoked back pain and then when it did not. Variability in the postural strategy of the abdominal muscles and pain-related cognitions were evaluated. Only those subjects for whom pain induced a reduction in variability of the postural strategy failed to return to a normal strategy when pain stopped. They were also characterized by their pain-related cognitions. Ongoing perception of threat to the back may exert tighter evaluative control over variability of the postural strategy.