Andy Weyer

Piezo2 is required for Merkel-cell mechanotransduction

How we sense touch remains fundamentally unknown. The Merkel cell–neurite complex is a gentle touch receptor in the skin that mediates slowly adapting responses of Aβ sensory fibres to encode fine details of objects. This mechanoreceptor complex was recognized to have an essential role in sensing gentle touch nearly 50 years ago. However, whether Merkel cells or afferent fibres themselves sense mechanical force is still debated, and the molecular mechanism of mechanotransduction is unknown. Synapse-like junctions are observed between Merkel cells and associated afferents, and yet it is unclear whether Merkel cells are inherently mechanosensitive or whether they can rapidly transmit such information to the neighbouring nerve. Here we show that Merkel cells produce touch-sensitive currents in vitro. Piezo2, a mechanically activated cation channel, is expressed in Merkel cells. We engineered mice deficient in Piezo2 in the skin, but not in sensory neurons, and show that Merkel-cell mechanosensitivity completely depends on Piezo2. In these mice, slowly adapting responses in vivo mediated by the Merkel cell–neurite complex show reduced static firing rates, and moreover, the mice display moderately decreased behavioural responses to gentle touch. Our results indicate that Piezo2 is the Merkel-cell mechanotransduction channel and provide the first line of evidence that Piezo channels have a physiological role in mechanosensation in mammals. Furthermore, our data present evidence for a two-receptor-site model, in which both Merkel cells and innervating afferents act together as mechanosensors. The two-receptor system could provide this mechanoreceptor complex with a tuning mechanism to achieve highly sophisticated responses to a given mechanical stimulus.

Involvement of Noradrenergic Neurotransmission in the Stress- but not Cocaine-Induced Reinstatement of Extinguished Cocaine-Induced Conditioned Place Preference in Mice: Role for β-2 Adrenergic Receptors

The responsiveness of central noradrenergic systems to stressors and cocaine poses norepinephrine as a potential common mechanism through which drug re-exposure and stressful stimuli promote relapse. This study investigated the role of noradrenergic systems in the reinstatement of extinguished cocaine-induced conditioned place preference by cocaine and stress in male C57BL/6 mice. Cocaine- (15 mg/kg, i.p.) induced conditioned place preference was extinguished by repeated exposure to the apparatus in the absence of drug and reestablished by a cocaine challenge (15 mg/kg), exposure to a stressor (6-min forced swim (FS); 20–25°C water), or administration of the α-2 adrenergic receptor (AR) antagonists yohimbine (2 mg/kg, i.p.) or BRL44408 (5, 10 mg/kg, i.p.). To investigate the role of ARs, mice were administered the nonselective β-AR antagonist, propranolol (5, 10 mg/kg, i.p.), the α-1 AR antagonist, prazosin (1, 2 mg/kg, i.p.), or the α-2 AR agonist, clonidine (0.03, 0.3 mg/kg, i.p.) before reinstatement testing. Clonidine, prazosin, and propranolol failed to block cocaine-induced reinstatement. The low (0.03 mg/kg) but not high (0.3 mg/kg) clonidine dose fully blocked FS-induced reinstatement but not reinstatement by yohimbine. Propranolol, but not prazosin, blocked reinstatement by both yohimbine and FS, suggesting the involvement of β-ARs. The β-2 AR antagonist ICI-118551 (1 mg/kg, i.p.), but not the β-1 AR antagonist betaxolol (10 mg/kg, i.p.), also blocked FS-induced reinstatement. These findings suggest that stress-induced reinstatement requires noradrenergic signaling through β-2 ARs and that cocaine-induced reinstatement does not require AR activation, even though stimulation of central noradrenergic neurotransmission is sufficient to reinstate.

Selective spider toxins reveal a role for the Nav1.1 channel in mechanical pain

Voltage-gated sodium (Nav) channels initiate action potentials in most neurons, including primary afferent nerve fibres of the pain pathway. Local anaesthetics block pain through non-specific actions at all Nav channels, but the discovery of selective modulators would facilitate the analysis of individual subtypes of these channels and their contributions to chemical, mechanical, or thermal pain. Here we identify and characterize spider (Heteroscodra maculata) toxins that selectively activate the Nav1.1 subtype, the role of which in nociception and pain has not been elucidated. We use these probes to show that Nav1.1-expressing fibres are modality-specific nociceptors: their activation elicits robust pain behaviours without neurogenic inflammation and produces profound hypersensitivity to mechanical, but not thermal, stimuli. In the gut, high-threshold mechanosensitive fibres also express Nav1.1 and show enhanced toxin sensitivity in a mouse model of irritable bowel syndrome. Together, these findings establish an unexpected role for Nav1.1 channels in regulating the excitability of sensory nerve fibres that mediate mechanical pain.

Transcriptional profiling at whole population and single cell levels reveals somatosensory neuron molecular diversity

The somatosensory nervous system is critical for the organisms ability to respond to mechanical, thermal, and nociceptive stimuli. Somatosensory neurons are functionally and anatomically diverse but their molecular profiles are not well-defined. Here, we used transcriptional profiling to analyze the detailed molecular signatures of dorsal root ganglion (DRG) sensory neurons. We used two mouse reporter lines and surface IB4 labeling to purify three major non-overlapping classes of neurons: 1) IB4+SNS-Cre/TdTomato+, 2) IB4−SNS-Cre/TdTomato+, and 3) Parv-Cre/TdTomato+ cells, encompassing the majority of nociceptive, pruriceptive, and proprioceptive neurons. These neurons displayed distinct expression patterns of ion channels, transcription factors, and GPCRs. Highly parallel qRT-PCR analysis of 334 single neurons selected by membership of the three populations demonstrated further diversity, with unbiased clustering analysis identifying six distinct subgroups. These data significantly increase our knowledge of the molecular identities of known DRG populations and uncover potentially novel subsets, revealing the complexity and diversity of those neurons underlying somatosensation. DOI: http://dx.doi.org/10.7554/eLife.04660.001

Pain Perception after Isometric Exercise in Women with Fibromyalgia

OBJECTIVE The purpose of this study was to identify exercise protocols incorporating isometric contractions that provide pain relief in women with fibromyalgia. DESIGN A before-after trial. SETTING A physical therapy department in an academic setting. PARTICIPANTS Fifteen women (mean ± SD, 52±11y) with fibromyalgia. INTERVENTIONS Subjects completed 4 sessions: 1 familiarization and 3 experimental. The following randomized experimental sessions involved the performance of isometric contractions with the elbow flexor muscles that varied in intensity and duration: (1) 3 maximal voluntary contractions (MVCs), (2) 25% MVC held to task failure, and (3) 25% MVC held for 2 minutes. MAIN OUTCOME MEASURES Experimental pain (pain threshold and pain rating), Fibromyalgia Impact Questionnaire, and fibromyalgia pain intensity (visual analog scale). RESULTS After all 3 isometric contractions, there was considerable variability between subjects in the pain response. Based on the changes in experimental pain, subjects were divided into 3 groups (increase, decrease, no change in pain). Multiple regression analysis revealed that age, baseline experimental pain, and change in fibromyalgia pain intensity were significant predictors of the experimental pain response after the isometric contractions. CONCLUSIONS We identified subgroups of women with fibromyalgia based on how they perceived pain after isometric contractions. The greatest pain relief for women with fibromyalgia occurred at a younger age and in women with the greatest experimental pain before exercise. Additionally, we established a link between experimental and clinical pain relief after the performance of isometric contractions.

Anxiety and stress can predict pain perception following a cognitive stress

Hoeger Bement, M.K., A. Weyer, M. Keller, A. Harkins, and S.K. Hunter. Anxiety and stress can predict pain perception following a cognitive stressor. PHYSIOL BEHAV 000-000. The purpose of this study was to investigate the influence of a cognitive stressor on pain perception and determine individual characteristics that may predict the pain response. Twenty-five subjects participated in three sessions: one familiarization and two experimental. The experimental sessions involved measurement of pain perception before and after 1) mental math tasks (stressor session) and 2) quiet rest (control session). Pain threshold and ratings were assessed with a mechanical noxious stimulus. Changes in stress and anxiety were examined with self-reported and physiological measures including questionnaires, visual analogue scales, and salivary cortisol levels. During the control session, stress and anxiety decreased and pain reports remain unchanged. During the stressor session, stress and anxiety increased and pain reports were variable among subjects. Based on the pain response to mental math, subjects were divided into three groups (increase, decrease or no change in pain). The increase-pain group (n=8) had lower baseline stress and anxiety, lower baseline pain reports, and large anxiety response following the mental math. In contrast, the decrease-pain group (n=9) had higher baseline stress and anxiety levels, higher baseline pain reports, and a large increase in cortisol levels. Thus, the differential response in the changes in pain perception was related to anxiety and stress levels prior to and during the cognitive stressor, indicating that psychosocial characteristics can help determine the stress-induced pain response.

Fatiguing exercise attenuates pain-induced corticomotor excitability

The purpose of this study was to assess net corticomotor excitability during a painful stimulus before and after exercise. In the first study, 25 subjects participated in three sessions: one familiarization session and two experimental sessions. The two experimental sessions were randomized and involved measurement of pain perception before and after (1) a submaximal isometric fatiguing contraction with the left elbow flexor muscles and (2) 30 min of quiet rest. Pain perception was assessed using a pressure device applied to the right index finger for 2 min. Motor evoked potentials (MEPs) of the left brachioradialis muscle were measured following transcranial magnetic stimulation (TMS) which was delivered before, during, and after the 2 min pain test. In the second study, 12 subjects participated in one session which involved application of TMS to elicit MEPs at the same time points as in study one, before and after a submaximal fatiguing contraction, but in the absence of pain. In the absence of the mechanical noxious stimulus, MEP amplitude was reduced following the fatiguing contraction and unchanged over this time period. In study one, pain threshold increased and pain ratings decreased following the isometric contraction but not after 30 min of quiet rest. During application of the mechanical noxious stimulus to the right index finger, MEP amplitude of the left brachioradialis muscle increased indicating an increase in net corticomotor excitability. The pain-induced increase in MEPs was attenuated following the isometric fatiguing contraction and this occurred in parallel with the decrease in pain.

A gain-of-function voltage-gated sodium channel 1.8 mutation drives intense hyperexcitability of A- and C-fiber neurons

Summary Gain‐of‐function mutation in Nav1.8 channel produces its most marked effect along the axon, resulting in intense spontaneous bursting and increased mechanically evoked action potentials in specific afferent subpopulations. ABSTRACT Therapeutic use of general sodium channel blockers, such as lidocaine, can substantially reduce the enhanced activity in sensory neurons that accompanies chronic pain after nerve or tissue injury. However, because these general blockers have significant side effects, there is great interest in developing inhibitors that specifically target subtypes of sodium channels. Moreover, some idiopathic small‐fiber neuropathies are driven by gain‐of‐function mutations in specific sodium channel subtypes. In the current study, we focus on one subtype, the voltage‐gated sodium channel 1.8 (Nav1.8). Nav1.8 is preferentially expressed in nociceptors, and gain‐of‐function mutations in Nav1.8 result in painful mechanical hypersensitivity in humans. Here, we used the recently developed gain‐of‐function Nav1.8 transgenic mouse strain, Possum, to investigate Nav1.8‐mediated peripheral afferent hyperexcitability. This gain‐of‐function mutation resulted in markedly increased mechanically evoked action potential firing in subclasses of A&bgr;, A&dgr;, and C fibers. Moreover, mechanical stimuli initiated bursts of action potential firing in specific subpopulations that continued for minutes after removal of the force and were not susceptible to conduction failure. Surprisingly, despite the intense afferent firing, the behavioral effects of the Nav1.8 mutation were quite modest, as only frankly noxious stimuli elicited enhanced pain behavior. These data demonstrate that a Nav1.8 gain‐of‐function point mutation contributes to intense hyperexcitability along the afferent axon within distinct sensory neuron subtypes.

Nociceptor Sensitization Depends on Age and Pain Chronicity

Abstract Peripheral inflammation causes mechanical pain behavior and increased action potential firing. However, most studies examine inflammatory pain at acute, rather than chronic time points, despite the greater burden of chronic pain on patient populations, especially aged individuals. Furthermore, there is disagreement in the field about whether primary afferents contribute to chronic pain. Therefore, we sought to evaluate the contribution of nociceptor activity to the generation of pain behaviors during the acute and chronic phases of inflammation in both young and aged mice. We found that both young (2 months old) and aged (>18 months old) mice exhibited prominent pain behaviors during both acute (2 day) and chronic (8 week) inflammation. However, young mice exhibited greater behavioral sensitization to mechanical stimuli than their aged counterparts. Teased fiber recordings in young animals revealed a twofold mechanical sensitization in C fibers during acute inflammation, but an unexpected twofold reduction in firing during chronic inflammation. Responsiveness to capsaicin and mechanical responsiveness of A-mechanonociceptor (AM) fibers were also reduced chronically. Importantly, this lack of sensitization in afferent firing during chronic inflammation occurred even as these inflamed mice exhibited continued behavioral sensitization. Interestingly, C fibers from inflamed aged animals showed no change in mechanical firing compared with controls during either the acute or chronic inflammatory phases, despite strong behavioral sensitization to mechanical stimuli at these time points. These results reveal the following two important findings: (1) nociceptor sensitization to mechanical stimulation depends on age and the chronicity of injury; and (2) maintenance of chronic inflammatory pain does not rely on enhanced peripheral drive.

Development of TRPM8 Antagonists to Treat Chronic Pain and Migraine

A review. Development of pharmaceutical antagonists of transient receptor potential melastatin 8 (TRPM8) have been pursued for the treatment of chronic pain and migraine. This review focuses on the current state of this progress.