Low-threshold Mechanical Stimuli Research Articles

Involvement of Mrgprd-expressing nociceptors-recruited spinal mechanisms in nerve injury-induced mechanical allodynia

Mechanical allodynia and hyperalgesia are intractable symptoms lacking effective clinical treatments in patients with neuropathic pain. However, whether and how mechanically responsive non-peptidergic nociceptors are involved remains elusive. Here, we showed that von Frey-evoked static allodynia and aversion, along with mechanical hyperalgesia after spared nerve injury (SNI) were reduced by ablation of MrgprdCreERT2-marked neurons. Electrophysiological recordings revealed that SNI-opened Aβ-fiber inputs to laminae I-IIo and vIIi, as well as C-fiber inputs to vIIi, were all attenuated in Mrgprd-ablated mice. In addition, priming chemogenetic or optogenetic activation of Mrgprd+ neurons drove mechanical allodynia and aversion to low-threshold mechanical stimuli, along with mechanical hyperalgesia. Mechanistically, gated Aβ and C inputs to vIIi were opened, potentially via central sensitization by dampening potassium currents. Altogether, we uncovered the involvement of Mrgprd+ nociceptors in nerve injury-induced mechanical pain and dissected the underlying spinal mechanisms, thus providing insights into potential therapeutic targets for pain management.

Cinnamaldehyde elicits itch behavior via TRPV1 and TRPV4 but not TRPA1.

Introduction:Cinnamaldehyde (CA) elicits itch sensation in humans. We investigated if CA elicits scratching behavior in mice and determined the roles for TRPV1, TRPA1, and TRPV4.Materials and Methods:Scratching behavior elicited by intradermal injection of CA was assessed in wildtype (WT) mice and knockout (KO) mice lacking TRPV1, TRPA1, TRPV4, or deficient in mast cells. We also assessed scratching and wet dog shakes elicited by low-threshold mechanical stimulation of skin treated topically with CA or vehicle. Using calcium imaging we tested if CA activates dorsal root ganglion (DRG) neurons of each genotype.Results:Intradermal cheek injection of CA elicited dose-dependent hindlimb scratch bouts, with fewer forelimb wipes and facial groom bouts that were not dose-dependent. CA elicited significantly fewer scratch bouts in TRPV1 and TRPV4 KO mice, but not TRPA1KOs, compared with WTs. There were no sex differences across genotypes. The histamine H1 antagonist cetirizine did not affect CA-evoked scratching, which was normal in mast cell deficient mice, indicating lack of histamine involvement. Scores for alloknesis were significantly greater following topical application of CA compared with vehicle. Post-CA alloknesis scores were significantly higher in TRPV4KOs of both sexes and in female TRPV1 and TRPA1KOs, compared with WTs. Low threshold mechanical stimuli also elicited significantly more wet dog shakes in mice treated topically with 20% CA, with significantly fewer in TRPV1, TRPA1, and TRPV4KOs compared with WTs. In calcium imaging studies, CA excited 24% of WT DRG cells, significantly fewer (11.5%) in cells from TRPV4KOs, and none in TRPA1KOs. Responses of cells of all genotypes exhibited significant sensitization to repeated CA stimulation. Sensitization was significantly enhanced by IL-4, which itself excited 16% of WT DRG cells and none from TRPA1KOs.Discussion:The results indicate that TRPA1 is dispensable for CA-evoked scratching, which depends partly on TRPV1 and TRPV4.

(350) The Persistent Pain Transcriptome: Distinct Transcriptional Signatures of Post-Surgical and Inflammatory Models Define Convergent Pathways to the Transition to Hyperalgesia

Most models of induced hyperalgesia display stereotyped behavioral responses to modalities such as heat and low threshold mechanical stimuli. Despite this, the nociceptive apparatus and the subsequent subjective experiences that define hyperalgesia differ greatly, as do accompanying symptoms of the associated pain state. Based on several recent in-depth studies of the activation patterns of primary afferents, we predicted that divergent activation paradigms could trigger distinct plastic events at second-order neurons that converge to sensitize the spinal circuits to evoked stimuli. The spinal circuits engaged by inflammatory and post-surgical pain states can be elucidated based on multiplex labeling and time course evaluations of the transcriptional signatures using deep sequencing. These experiments revealed a population of neurons apparently undetected by single-cell and single-nucleus sequencing methods because of its sparse representation in the superficial laminae of the dorsal horn. Investigations into the character and connectivity of these second order spinal neurons revealed that they participate similarly in at least two models of hyperalgesia. These models are driven by different transcriptional machinery at the level of the dorsal spinal cord, and the most transcriptionally evident neurons express very high levels of both the opioid peptide dynorphin and receptors that may be involved in phenotypic switching during hyperalgesia. We identified multiple cell populations, representing opponent processes acting to promote and abrogate hyperalgesia. Surprisingly, both neural and glial substrates of plasticity were divergent between the models, but converged at the pathway level. Examples of convergent pathways included immune-like activation and upregulation of the dynorphin precursor gene, whereas the activation of neuronal genes was divergent. These investigations outline a multidisciplinary strategy for identification of targets for analgesia and hyperalgesia research, and identify a set of neuronal receptors for future analgesic target development.

1055 Evaluation of film dressings for the prevention of intraepidermal nerve growth and alloknesis (touch-evoked itch) in murine dry skin models

Epidermal hyperinnervation is observed in dry skin diseases such as xerosis and atopic dermatitis. Such diseases manifest in low-threshold mechanical stimuli irritation of the skin, which is known as alloknesis. Increased intraepidermal nerve fibers are thought to be one of the pathogenic factors in itch hypersensitivity. Film dressings for wound treatment act as a skin barrier and provide a moist environment. However, it has not been clear whether film dressings have any preventive effects on epidermal hyperinnervation and alloknesis in dry skin diseases. We evaluated two different thickness film dressings, TEGADERM (TDM) and PERME ROLL Lite (PMR), for their preventive effect on epidermal hyperinnervation using an acute dry skin model. Twenty square millimeters of TDM or PMR was applied after 5-min acetone treatment to the back skin of ICR mice. Skin samples were taken after an additional 48 h. Immunohistochemically, the numbers of protein gene product 9.5-immunoreactive nerve fibers in and penetrating into the epidermis were significantly decreased by TDM or PMR treatment. We further evaluated alloknesis using a chronic dry skin model. A mixture of acetone and diethylether (1:1) for 15 s followed by distilled water for 30 s (AEW) was applied to the back skin of C57BL/6NCrSlc mice twice daily for 8 days. TDM or PMR with a diameter of 8 mm was applied on the AEW-treated site on the day after the last treatment. Alloknesis assays showed that TDM and PMR application significantly prevented induction of alloknesis. These findings suggest that the film dressings not only prevent epidermal hyperinnervation by moisturization but also reduce alloknesis by inhibiting the transmission of mechanical stimuli.

CaMKII Controls Whether Touch Is Painful.

The sensation of touch is served by specialized sensory neurons termed low-threshold mechanoreceptors (LTMRs). We examined the role of CaMKII in regulating the function of these neurons. Loss of CaMKII function, such as occurred in rats during sensory deprivation, elevated the generation and propagation of impulses by LTMRs, and altered the spinal cord circuitry in such a way that low-threshold mechanical stimuli produced pain behavior. Because limbs are protected from use during a painful condition, this sensitization of LTMRs may perpetuate pain and prevent functional rehabilitation.

Nalfurafine Suppresses Pruritogen- and Touch-evoked Scratching Behavior in Models of Acute and Chronic Itch in Mice

The kappa-opioid agonist, nalfurafine, has been approved in Japan for treatment of itch in patients with chronic kidney disease. We presently investigated if systemic administration of nalfurafine inhibited ongoing or touch-evoked scratching behavior (alloknesis) following acute intradermal injection of histamine or the non-histaminergic itch mediator, chloroquine, in mice. We also investigated if nalfurafine suppressed spontaneous or touch-evoked scratching in an experimental model of chronic dry skin itch. Nalfurafine reduced scratching evoked by histamine and chloroquine. Following acute histamine, but not chloroquine, low-threshold mechanical stimuli reliably elicited directed hindlimb scratching behavior, which was significantly attenuated by nalfurafine. In mice with experimental dry skin, nalfurafine abolished spontaneous scratching but had no effect on alloknesis. Nalfurafine thus appears to be a promising treatment for acute itch as well as ongoing itch of dry skin.

Subclinical peripheral neuropathy in patients with multiple myeloma before chemotherapy is correlated with decreased fingertip innervation density.

The goal in this study was to determine the incidence of subclinical neuropathy in treatment-naive patients with multiple myeloma (MM) with no history of peripheral neuropathy using quantitative sensory tests (QSTs) and its correlation with innervation density of the extremities using noninvasive laser reflectance confocal microscopy. QST results were collected for 27 patients with a diagnosis of MM and compared with data collected from 30 age- and sex-matched healthy volunteers. Skin temperature, sensorimotor function (grooved pegboard test), and detection thresholds for temperature, sharpness, and low-threshold mechanical stimuli (von Frey monofilaments and bumps detection test) were measured. Meissner's corpuscle (MC) density in the fingertips was assessed using in vivo laser reflectance confocal microscopy. Patients showed a high incidence (> 80%) of ≥ one subclinical QST deficit. These included increased von Frey, bumps, and warmth detection thresholds as compared with healthy volunteers. Patients also showed increases in cold pain, sensorimotor deficits (grooved pegboard test), and higher overall neuropathy scores. MC density was significantly lower in patients than controls and showed significant inverse correlation with bumps detection threshold. Patients with MM commonly present with sensory and sensorimotor deficits before undergoing treatment, and these deficits seem to result from disease-related decreases in peripheral innervation density.

Behavioral model of itch, alloknesis, pain and allodynia in the lower hindlimb and correlative responses of lumbar dorsal horn neurons in the mouse

We have further developed a behavioral model of itch and pain in the lower hindlimb (calf) originally reported by LaMotte et al. (2011) that allows comparisons with responses of lumbar dorsal horn neurons to pruritic and noxious stimuli. Intradermal (id) microinjection of the pruritogens histamine, SLIGRL-NH2 (agonist of PAR-2 and MrgprC11) and chloroquine (agonist of MrgprA3) into the calf of the lower limb elicited significant biting and a small amount of licking directed to the injection site, over a 30-min time course. Following id injection of histamine, low-threshold mechanical stimuli reliably elicited discrete episodes of biting (alloknesis) over a longer time course; significantly less alloknesis was observed following id injection of SLIGRL-NH2. Capsaicin injections elicited licking but little biting. Following id injection of capsaicin, low-threshold mechanical stimuli elicited discrete hindlimb flinches (allodynia) over a prolonged (>2h) time course. In single-unit recordings from superficial lumbar dorsal horn neurons, low-threshold mechanically evoked responses were significantly enhanced, accompanied by receptive field expansion, following id injection of histamine in histamine-responsive neurons. This was not observed in histamine-insensitive neurons, or following id injection of saline or SLIGRL-NH2, regardless of whether the latter activated the neuron or not. These results suggest that itch-responsive neurons are selectively sensitized by histamine but not SLIGRL-NH2 to account for alloknesis. The presently described “calf” model appears to distinguish between itch- and pain-related behavioral responses, and provides a basis to investigate lumbar spinal neural mechanisms underlying itch, alloknesis, pain and allodynia.

Eugenol and carvacrol induce temporally desensitizing patterns of oral irritation and enhance innocuous warmth and noxious heat sensation on the tongue

Eugenol and carvacrol, from the spices clove and oregano, respectively, are agonists of TRPV3, which is implicated in transduction of warmth and possibly heat pain. We investigated the temporal dynamics of lingual irritation elicited by these agents, and their effects on innocuous warmth and heat pain, using a half-tongue method in human subjects. The irritant sensation elicited by both eugenol and carvacrol decreased across repeated applications at a 1-minute interstimulus interval (self-desensitization) which persisted for at least 10minutes. Both agents also cross-desensitized capsaicin-evoked irritation. Eugenol and carvacrol significantly increased the magnitude of perceived innocuous warmth (44°C) for >10minutes, and briefly (<5minutes) enhanced heat pain elicited by a 49°C stimulus. Similar albeit weaker effects were observed when thermal stimuli were applied after the tongue had been desensitized by repeated application of eugenol or carvacrol, indicating that the effect is not due solely to summation of chemoirritant and thermal sensations. Neither chemical affected sensations of innocuous cool or cold pain. A separate group of subjects was asked to subdivide eugenol and carvacrol irritancy into subqualities, the most frequently reported being numbing and warmth, with brief burning, stinging/pricking, and tingle, confirming an earlier study. Eugenol, but not carvacrol, reduced detection of low-threshold mechanical stimuli. Eugenol and carvacrol enhancement of innocuous warmth may involve sensitization of thermal gating of TRPV3 expressed in peripheral warm fibers. The brief heat hyperalgesia following eugenol may involve a TRPV3-mediated enhancement of thermal gating of TRPV1 expressed in lingual polymodal nociceptors.

Discriminative sensory characteristics of the lateral femoral cutaneous nerve after mepivacaine-induced block

Background and objectivesUnmyelinated C-fibres comprise the largest group of somatic afferents and have demonstrated a crucial role not only in the perception of high-threshold mechanically, thermally or chemically induced pain, but also in non-harmful low-threshold mechanical stimuli [1,2]. The objective of our study was to characterize differential sensitivity changes of C-fibre related subclasses of high-threshold and low-threshold polymodal nociceptors and low-threshold mechanoreceptors to the local anaesthetic (LA) mepivacaine during nerve block of the purely sensory lateral femoral cutaneous nerve (LFCN) in human. We assumed a diverse response of different classes of afferents to the two different concentrations of the LA mepivacaine (Scandicaine). MethodsIn a double-blind randomized experimental setting, an ultrasound-guided nerve block of the LFCN was performed in 10 healthy male subjects, each with two different concentrations of mepivacaine (0.5 and 1%). Responsiveness of afferent nerve fibres to different noxious and non-noxious stimuli was tested by Quantitative Sensory Testing (QST) 30, 180, and 300min after nerve block. Both LA concentrations of mepivacaine were compared for time course of the areas of anaesthesia for the tested sensory modalities. ResultsInitial extension of anaesthetic areas at 30min did not differ between both LA concentrations. At 180min only the anaesthetic areas to nociceptive stimuli were reduced at the site of lower mepivacaine injection (260mN: 204mm2 (18; 244; median difference and 95% confidence interval; p<0.05), heat: 276mm2 (3; 305)). In contrast, no significant differences were found between the two concentration when non-nociceptive stimuli were used (100mN: 187mm2 (4; 240), p>0.05, brush: 159mm2 (−59; 202)). ConclusionEqual initial sizes of anaesthesia areas for all sensory modalities can be explained by supramaximal perineural LA molecule concentration in both administered mepivacaine dosages. Upon washout of the LA nociceptive function is restored faster as compared to non-nociceptive sensation and higher concentration of the LA are required to maintain the analgesia. Quantitative sensory testing is able to detect different susceptibility of low threshold mechanosensors and subtypes of nociceptive C-fibres to mepivacaine. Using painful mechanical, heat and electrical stimulation different classes of nociceptors will be activated. The analgesic areas to electrical stimulation were particularly small; one might therefore hypothesize that the proposed protocol allows to also differentiate mechano-insensitive (“silent”) and mechanosensitive (“polymodal”) nociceptors. ImplicationsQST is a non-invasive method to functionally examine sensory modalities and their pharmacological modulation in humans. The method is sufficiently sensitive to differentiate the analgesic properties of mepivacaine at 0.5 and 1% and might also be adequate to different classes of nociceptors. Further development of nociceptive stimuli including supra-threshold encoding characteristics will enable to investigate peripheral analgesic effects more specifically and thus might help to design new analgesics with preferential effect on high frequency discharge of nociceptors.

Light touch induces ERK activation in superficial dorsal horn neurons after inflammation: involvement of spinal astrocytes and JNK signaling in touch‐evoked central sensitization and mechanical allodynia

Activation of extracellular signal-regulated kinase (ERK) in spinal cord neurons could serve as a marker for sensitization of dorsal horn neurons in persistent pain. ERK is normally activated by high-threshold noxious stimuli. We investigated how low-threshold mechanical stimuli could activate ERK after complete Freund's adjuvant (CFA)-induced inflammation. Unilateral injection of CFA induced ipsilateral heat hyperalgesia and bilateral mechanical allodynia. CFA-induced ERK activation in ipsilateral dorsal horn neurons declined after 2 days. Interestingly, low-threshold mechanical stimulation given by light touch either on the inflamed paw or the contralateral non-inflamed paw dramatically increased ERK phosphorylation in the dorsal horn ipsilateral to touch stimulation. Notably, light touch induced ERK phosphorylation mainly in superficial neurons in laminae I-IIo. Intrathecal administration of the astroglial toxin L-α-aminoadipate on post-CFA day 2 reversed CFA-induced bilateral mechanical allodynia but not heat hyperalgesia. Furthermore, L-α-aminoadipate, the glial inhibitor fluorocitrate, and a peptide inhibitor of c-Jun N-terminal Kinase all reduced light touch-evoked ERK activation ipsilateral to touch. Collectively, these data suggest that (i) ERK can be activated in superficial dorsal horn neurons by low-threshold mechanical stimulation under pathological condition and (ii) ERK activation by light touch is associated with mechanical allodynia and requires an astrocyte network.

Transient Receptor Potential Vanilloid-4 Has a Major Role in Visceral Hypersensitivity Symptoms

The transient receptor potential vanilloid-4 (TRPV4) is an osmosensitive channel that responds to mechanical stimulation. We hypothesized that TRPV4 could be important in visceral nociception and in the development of hypersensitivity. TRPV4 expression was investigated by immunohistochemistry and reverse transcription-polymerase chain reaction. Calcium signaling and patch-clamp studies were performed in dorsal root ganglia (DRG) neurons validating the use of 4alphaPDD as a selective TRPV4 agonist. The effects of TRPV4 activation on visceral nociception were evaluated in mice that received intracolonically TRPV4 agonist (4 alpha-phorbol 12,13-didecanoate [4alphaPDD]) and in TRPV4-deficient mice in which abdominal muscle contractions in response to colorectal distention (CRD) were recorded. Intervertebral injections of TRPV4 or mismatch small interfering RNA (siRNA) were used to specifically down-regulate TRPV4 expression in sensory neurons and to investigate the role of TRPV4 in basal visceral nociception or in protease-activated receptor 2 (PAR(2)) activation-induced visceral hypersensitivity. TRPV4 agonist 4alphaPDD specifically activated a cationic current and calcium influx in colonic projections of DRG neurons and caused dose-dependent visceral hypersensitivity. TRPV4-targeted but not mismatched siRNA intervertebral treatments were effective at reducing basal visceral nociception, as well as 4alphaPDD or PAR(2) agonist-induced hypersensitivity. Effects of the TRPV4 ligand were lost in TRPV4-deficient mice. 4alphaPDD selectively activates TRPV4 in sensory neurons projecting from the colon, and TRPV4 activation causes visceral hypersensitivity. TRPV4 activation is implicated in the nociceptive response to CRD in basal conditions and in PAR(2) agonist-induced hypersensitivity. These results suggest a pivotal role for TRPV4 in visceral nociception and hypersensitivity.

TREK-1, a K+ channel involved in polymodal pain perception

The TREK-1 channel is a temperature-sensitive, osmosensitive and mechano-gated K+ channel with a regulation by Gs and Gq coupled receptors. This paper demonstrates that TREK-1 qualifies as one of the molecular sensors involved in pain perception. TREK-1 is highly expressed in small sensory neurons, is present in both peptidergic and nonpeptidergic neurons and is extensively colocalized with TRPV1, the capsaicin-activated nonselective ion channel. Mice with a disrupted TREK-1 gene are more sensitive to painful heat sensations near the threshold between anoxious warmth and painful heat. This phenotype is associated with the primary sensory neuron, as polymodal C-fibers were found to be more sensitive to heat in single fiber experiments. Knockout animals are more sensitive to low threshold mechanical stimuli and display an increased thermal and mechanical hyperalgesia in conditions of inflammation. They display a largely decreased pain response induced by osmotic changes particularly in prostaglandin E2-sensitized animals. TREK-1 appears as an important ion channel for polymodal pain perception and as an attractive target for the development of new analgesics.

Statistical Modeling of the Response Characteristics of Mechanosensitive Stimuli in the Human Esophagus

It is believed that mechanical stimuli of the human gut activate afferents responding to either noxious or normal, physiologic stimuli. They might be able to sensitize without relation to the contractile state of the smooth muscle. The current study aimed to verify the above characteristics by using a statistical model based on correlation analysis. The esophagus was distended with a bag in 32 healthy subjects by using an inflation rate of 25 mL/min. The luminal cross-sectional areas and sensory ratings were determined during the distentions. The stimuli were repeated after relaxation of the smooth muscle with butylscopolamine and after sensitization with hydrochloride acid. A positive correlation between the sensory responses to distention was found in the nonpainful and painful ranges, respectively, but correlations between nonpainful and painful ratings were nonsignificant. Relaxation of the smooth muscle did not influence the correlations, and sensitization resulted in inter-individual differences and disappearance of the above clustering into painful and nonpainful correlations. In conclusion, afferent nerves encoding high-threshold and low-threshold mechanical stimuli of the human esophagus are not correlated and thus probably represent different populations. The response characteristics have no physiologic relationship to the contractile state of the smooth muscle, and sensitization affects all types of afferents. Perspectives The article adds information about sensory processing of mechanical gut stimuli in human beings. This might increase our understanding of visceral pain in health and disease and guide the statistical analysis of experimental data obtained in the gastrointestinal tract.

Dendritic architecture of rat somatosensory thalamocortical projection neurons

This study examines dendrites from physiologically characterized and intracellularly labelled thalamocortical projection (TCP) neurons from the rat ventrobasal complex (VB) and posterior nucleus (POm). The goals were to provide quantitative descriptions of TCP neuron dendrites, examine underlying design principles of dendritic morphology, and determine correlations between dendritic size parameters. Forty-four dendrites from seven VB neurons and 21 dendrites from three POm TCP neurons that responded to low-threshold mechanical stimuli were reconstructed and quantitatively analyzed at the light microscopic level. The dendritic architecture of the neurons was remarkably similar in most parameters studied, including the percentage of dichotomous branching, contribution of terminal branches to total dendritic length, and branching symmetry. There was a positive correlation between stem dendrite diameter and the length of the entire dendrite arbor, making it possible to estimate the total length of a dendritic arbor by measuring the stem dendrite diameter. The correlations of the VB and POm dendrites had different slopes. The path distance (the distance from the soma to a dendritic end point) of individual dendrites showed only a small variation with large differences in the total dendritic length of an arbor. The constant diameter of distal dendrites shows that dendrite diameter is a poor predictor of synaptic location on the dendritic tree. Although the morphology of neurons and their individual dendrites varied considerably in overall size and qualitative appearance, when examined qualitatively, many aspects of dendritic structure were similar within and between groups. We suggest that the rat somatosensory TCP neurons have a stereotyped dendritic architecture and present data which provide a base for future comparative, developmental, and plasticity studies.

Properties of feline thalamic neurons activated by stimulation of the middle meningeal artery and sagittal sinus

Previous studies have identified a population of neurons in the cat trigeminal brainstem complex that respond to stimulation of the middle miningeal artery (MMA) and/or superior sagittal sinus (SS). In the present study, neuronal responses to stimulation of the MMA and SS were sought in the thalamus of the cat. Sixty-one neurons excited by electrical stimulation of the MMA and/or SS were located in the ventroposteromedial (VPM) nucleus, and surrounding regions in lateral thalamus. Of these 61 neurons, 23% were excited only by MMA stimulation, 39% only by SS stimulation and 38% by both MMA and SS stimulation. The latencies to activation from MMA and/or SS stimulation suggest the involvement of small, myelinated primary afferent fibers. Most neurons (48/61) responded to electrical stimulation of the MMA or SS with a burst of 2–5 spikes. Mechanical stimulation of the MMA and SS was also an effective stimulus and in some cases evoked a burst response. Of the neurons tested for the existence of orofacial inputs, all were found to have an excitatory receptive field (RF) on the face and usually (22/26 neurons) involved the opthalmic distribution. Twelve of these neurons were excited by a tap stimulus applied to the face, 6 by pinching (nociceptive specific), 6 by low-threshold mechanical stimuli (LTM), one by both pinch and low-threshold stimuli and one by mechanical stimulation of the cornea. The firing properties, RF and modalities of these thalamic neurons suggest that they may play a role in the appreciation of pain of cerebrovascular origin.

Closely coupled excitation of gamma-motoneurones by group III Muscle afferents with low mechanical threshold in the cat.

1. The reflex responses of gamma-motoneurones to discharges of muscle receptors innervated by Group III axons have been examined in hind-limb muscles of decerebrated and spinal cats.2. Electrical stimulation of the gastrocnemius medialis nerve at a strength sufficient to excite Group III axons caused excitation of gastrocnemius lateralis or soleus gamma-motoneurones. The excitation was more prominent in the spinal animal.3. Excitation of either silent or tonically firing gamma-motoneurones was a secure, driven type of response consisting of one or two spikes occurring at fairly fixed latency in response to a single stimulus. Eighteen out of thirty-nine gamma-motoneurones studied showed such excitation.4. Subtracting peripheral conduction times of the gamma-motoneurone impulse and the earliest component of the Group III volley from the latency of reflex excitation gave a range of central delays of 1.8-4.8 msec (mean 3.0 msec).5. Inhibitory Group III effects were also seen but were less pronounced than the excitation.6. The origin of the receptors connected to the Group III axons whose discharge causes driven excitation of gamma-motoneurones was investigated.7. Isometric twitch contractions of a muscle caused pronounced facilitation or excitation of homonymous and heteronymous gamma-motoneurones. Excitation occurred close to the peak or during relaxation of the twitch with a range in latency of 30-110 msec. The muscles studied were triceps surae, flexor digitorum and hallucis longus.8. Pressure or light taps applied to the gastrocnemius medialis muscle caused an increase in discharge frequency in twenty out of twenty-eight gamma-motoneurones of the same muscle (four were inhibited). The response to a steady stimulus adapted but could outlast it by many seconds. Pressure was more effective when applied to the proximal or distal parts of the muscle.9. Fifteen out of thirty-eight gastrocnemius medialis receptors having Group III axons (conduction velocities 5-27 m/sec) were found to discharge one, or occasionally two, spikes to twitch contractions of the parent muscle at latencies of 30-80 msec.10. Eight out of ten receptors with Group III axons that responded to contraction had low thresholds to pressure and taps, whereas the majority of the twenty-three Group III units which were insensitive to contraction had high thresholds to such stimuli. Receptive fields of all Group III units were confined to either the proximal third or distal third of gastrocnemius medialis or, in a few instances, the Achilles tendon.11. We conclude that discharges in Group III axons from receptors which respond to non-noxious, low threshold mechanical stimuli cause a tightly coupled excitation of gamma-motoneurones. The likely contribution of this reflex to the control of movement is discussed.