Low-threshold Mechanoreceptive Units Research Articles

Electrophysiological differences between nociceptive and non-nociceptive dorsal root ganglion neurones in the rat in vivo.

Intracellular recordings were made from 1022 somatic lumbar dorsal root ganglion (DRG) neurones in anaesthetized adult rats, classified from dorsal root conduction velocities (CVs) as C, Adelta or Aalpha/beta, and according to their responses to mechanical and thermal stimuli as nociceptive (including high-threshold mechanoreceptive (HTM) units), and non-nociceptive (including low-threshold mechanoreceptive (LTM) and cooling units). Of these, 463 met electrophysiological criteria for analysis of action potentials (APs) evoked by dorsal root stimulation. These included 47 C-, 71 Adelta- and 102 Aalpha/beta-nociceptive, 10 C-, 8 Adelta- and 178 Aalpha/beta-LTM, 18 C- and 19 Adelta- unresponsive, and 4 C-cooling units. Medians of AP and afterhyperpolarization (AHP) durations and AP overshoots were significantly greater for nociceptive than LTM units in all CV groups. AP overshoots and AHP durations were similar in nociceptors of all CV groups whereas AP durations were greater in slowly conducting, especially C-fibre, nociceptors. C-cooling units had faster CVs, smaller AP overshoots and shorter AP durations than C-HTM units. A subgroup of Aalpha/beta-HTM, moderate pressure units, had faster CVs and AP kinetics than other Aalpha/beta-HTM units. Of the Aalpha/beta-LTM units, muscle spindle afferents had the fastest CV and AP kinetics, while rapidly adapting cutaneous units had the slowest AP kinetics. AP variables in unresponsive and nociceptive units were similar in both C- and Adelta-fibre CV groups. The ability of fibres to follow rapid stimulus trains (fibre maximum following frequency) was correlated with CV but not sensory modality. These findings indicate both the usefulness and limitations of using electrophysiological criteria for identifying neurones acutely in vitro as nociceptive.

The TTX-resistant sodium channel Nav1.8 (SNS/PN3): expression and correlation with membrane properties in rat nociceptive primary afferent neurons.

We have examined the distribution of the sensory neuron-specific Na+ channel Nav1.8 (SNS/PN3) in nociceptive and non-nociceptive dorsal root ganglion (DRG) neurons and whether its distribution is related to neuronal membrane properties. Nav1.8-like immunoreactivity (Nav1.8-LI) was examined with an affinity purified polyclonal antiserum (SNS11) in rat DRG neurons that were classified according to sensory receptive properties and by conduction velocity (CV) as C-, Adelta- or Aalpha/beta. A significantly higher proportion of nociceptive than low threshold mechanoreceptive (LTM) neurons showed Nav1.8-LI, and nociceptive neurons had significantly more intense immunoreactivity in their somata than LTM neurons. Results showed that 89, 93 and 60% of C-, Adelta- and Aalpha/beta-fibre nociceptive units respectively and 88% of C-unresponsive units were positive. C-unresponsive units had electrical membrane properties similar to C-nociceptors and were considered to be nociceptive-type neurons. Weak positive Nav1.8-LI was also present in some LTM units including a C LTM, all Adelta LTM units (D hair), about 10% of cutaneous LTM Aalpha/beta-units, but no muscle spindle afferent units. Nav1.8-LI intensity was negatively correlated with soma size (all neurons) and with dorsal root CVs in A- but not C-fibre neurons. Nav1.8-LI intensity was positively correlated with action potential (AP) duration (both rise and fall time) in A-fibre neurons and with AP rise time only in positive C-fibre neurons. It was also positively correlated with AP overshoot in positive neurons. Thus high levels of Nav1.8 protein may contribute to the longer AP durations (especially in A-fibre neurons) and larger AP overshoots that are typical of nociceptors.

Sensory and electrophysiological properties of guinea-pig sensory neurones expressing Nav 1.7 (PN1) Na+ channel alpha subunit protein.

The TTX-sensitive Na(v)1.7 (PN1) Na(+) channel alpha subunit protein is expressed mainly in small dorsal root ganglion (DRG) neurones. This study examines immunocytochemically whether it is expressed exclusively or preferentially in nociceptive primary afferent DRG neurones, and determines the electrophysiological properties of neurones that express it. Intracellular somatic action potentials (APs) evoked by dorsal root stimulation were recorded in L6/S1 DRG neurones at 30 +/- 2 degrees C in vivo in deeply anaesthetised young guinea-pigs. Each neurone was classified, from its dorsal root conduction velocity (CV) as a C-, Adelta- or Aalpha/beta-fibre unit and from its response to mechanical and thermal stimuli, as a nociceptive, low threshold mechanoreceptive (LTM) or unresponsive unit. Fluorescent dye was injected into the soma and Na(v)1.7-like immunoreactivity (Na(v)1.7-LI) was examined on sections of dye-injected neurones. All C-, 90 % of Adelta- and 40 % of Aalpha/beta-fibre units, including both nociceptive and LTM units, showed Na(v)1.7-LI. Positive units included 1/1 C-LTM, 6/6 C-nociceptive, 4/4 C-unresponsive (possible silent nociceptive) units, 5/6 Adelta-LTM (D hair), 13/14 Adelta-nociceptive, 2/9 Aalpha/beta-nociceptive, 10/18 Aalpha/beta-LTM cutaneous and 0/9 Aalpha/beta-muscle spindle afferent units. Overall, a higher proportion of nociceptive than of LTM neurones was positive, and the median relative staining intensity was greater in nociceptive than LTM units. Na(v)1.7-LI intensity was clearly positively correlated with AP duration and (less strongly) negatively correlated with CV and soma size. Since nociceptive units tend overall to have longer duration APs, slower CVs and smaller somata, these correlations may be related to the generally greater expression of Na(v)1.7 in nociceptive units.

The Presence and Role of the Tetrodotoxin-Resistant Sodium Channel Nav1.9 (NaN) in Nociceptive Primary Afferent Neurons

This is the first examination of sensory receptive properties and associated electrophysiological properties in vivo of dorsal root ganglion (DRG) neurons that express the TTX-resistant sodium channel Na(v)1.9 (NaN). Intracellular recordings in lumbar DRGs in Wistar rats enabled units with dorsal root C-, Adelta-, or Aalpha/beta-fibers to be classified as nociceptive, low-threshold mechanoreceptive (LTM), or unresponsive. Intracellular dye injection enabled subsequent immunocytochemistry for Na(v)1.9-like immunoreactivity (Na(v)1.9-LI). Na(v)1.9-LI was expressed selectively in nociceptive-type (C- and A-fiber nociceptive and C-unresponsive) units. Of the nociceptive units, 64, 54, and 31% of C-, Adelta-, and Aalpha/beta-fiber units, respectively, were positive for Na(v)1.9-LI. C-unresponsive units were included in the nociceptive-type group on the basis of their nociceptor-like membrane properties; 91% were positive. Na(v)1.9-LI was undetectable in Adelta- or Aalpha/beta-fiber LTM units and in one C-LTM unit. Na(v)1.9-LI intensity was correlated negatively with soma size and conduction velocity in nociceptive units and with conduction velocity in C-fiber units. There was a positive correlation with action potential rise time in nociceptive-type units with membrane potentials equal to or more negative than -50 mV. The data provide direct evidence that Na(v)1.9 is expressed selectively in (but not in all) C- and A-fiber nociceptive-type units and suggest that Na(v)1.9 contributes to membrane properties that are typical of nociceptive neurons.

Calcitonin gene-related peptide immunoreactivity and afferent receptive properties of dorsal root ganglion neurones in guinea-pigs.

To establish the afferent receptive properties of lumbosacral dorsal root ganglion (DRG) neurones that express calcitonin gene-related peptide (CGRP), intracellular recordings were made with fluorescent dye-filled electrodes in deeply anaesthetised young guinea-pigs. After determination of neuronal functional properties, dye was injected into the soma. CGRP-like immunoreactivity (CGRP-LI) was examined on histological sections of dye-marked neurones. Fourteen of 34 C-fibre neurones showed CGRP-LI. These included 10/21 C-fibre nociceptive neurones. All C-polymodal nociceptors in glabrous (n = 4) but none in hairy skin (n = 4) were positive. Positive C-fibre high threshold mechanoreceptive (HTM) units had receptive fields in dermal or deeper tissue. Four (n = 6) unresponsive or unidentified C-fibre units were positive. Neither C-fibre cooling sensitive (n = 4) nor C-fibre low threshold mechanoreceptive (LTM) units (n = 3) had CGRP-LI. Six of 23 A-fibre nociceptive cells were positive including one Aalpha/beta unit. Three of these positive cells had epidermal and three had dermal/deep receptive fields. Three of 36 A-fibre LTM units exhibited CGRP-LI; all were Aalpha/beta-fibre G hair units. All glabrous skin and muscle spindle units and in hairy skin slowly adapting and field units, and some G-hair units lacked CGRP-LI. CGRP-LI stained fibres were found in tissues containing receptive fields of positive DRG neurones: glabrous skin, near hair follicles and in skeletal muscle. A few substance P-labelled neurones did not exhibit CGRP-LI and vice versa. Thus CGRP expression was detected in under half the nociceptive neurones, was not limited to nociceptive neurones and apart from receptive properties was also related to location/depth in the tissues of a DRG neurone's peripheral terminals.

Phenotype and Function of Somatic Primary Afferent Nociceptive Neurones with C-, Aδ- or Aα/β-Fibres.

Nociceptive dorsal root ganglion (DRG) neurones have fibres that conduct in the C, Aδ and Aα/β conduction velocity range. The properties of nociceptive compared with non-nociceptive somatic afferent dorsal root ganglion neurones appear to fall into two patterns, A and B. Pattern A properties of nociceptive neurones, the more common type, include longer action potential duration and slower maximum rate of fibre firing, as well as a greater expression of substance P and calcitonin gene-related peptide immunoreactivity. The values of pattern A properties appear to be graded according to the conduction velocity group (C, Aδ or Aα/β) of the fibres. The most pronounced forms of A-type properties are expressed by nociceptive neurones with C-fibres, and these become less pronounced in nociceptive neurones with Aδ-fibres and least pronounced in those with Aα/β fibres (C > Aδ > Aα/β). Some of these properties are also expressed in a less extreme but similarly graded manner through C, Aδ and Aα/β groups of non-nociceptive low threshold mechanoreceptive (LTM) neurone. The less common pattern B properties of nociceptive neurones have similar values in C-, Aδ- and Aα/β-fibre nociceptive neurones but these clearly differ from LTM units with C-, Aδ- and Aα/β-fibre conduction velocities. These features of nociceptive neurones include consistently larger action potential overshoots and longer after-hyperpolarisation durations in nociceptive than in LTM neurones.

Association of somatic action potential shape with sensory receptive properties in guinea-pig dorsal root ganglion neurones.

1. Intracellular voltage recordings were made from the somata of L6 and S1 dorsal root ganglion (DRG) neurones at 28.5-31 C in young guinea-pigs (150-300 g) anaesthetized with sodium pentobarbitone. Action potentials (APs) evoked by dorsal root stimulation were used to classify conduction velocities (CVs) as C, Adelta or Aalpha/beta. Units with overshooting APs and membrane potentials (Vm) more negative than -40 mV were analysed: 40 C-, 45 Adelta- and 94 Aalpha/beta-fibre units. 2. Sensory receptive properties were characterized as: (a) low-threshold mechanoreceptive (LTM) units (5 C-, 10 Adelta- and 57 Aalpha/beta-fibre units); (b) nociceptive units, responding to noxious mechanical stimuli, some also to noxious heat (40 C-, 27 Adelta- and 27 Aalpha/beta-fibre units); (c) unresponsive units that failed to respond to a variety of tests; and (d) C-fibre cooling-sensitive units (n = 4). LTM units made up about 8 % of identified C-fibre units, 36 % of identified Adelta-fibre units and > 73 % of identified Aalpha/beta-fibre units. Compared with LTM units, the nociceptive units had APs that were longer on average by 3 times (C-fibre units), 1.7 times (Adelta-fibre units) and 1.4 times (Aalpha/beta-fibre units). They also had significantly longer rise times (RTs) and fall times (FTs) in all CV ranges. Between Aalpha/beta-nociceptors and Aalpha/beta-LTMs there was a proportionately greater difference in RT than in FT. The duration of the afterhyperpolarization measured to 80 % recovery (AHP80) was also significantly longer in nociceptive than LTM neurones in all CV ranges: by 3 times (C-fibre units), 6.3 times (Adelta-fibre units) and 3.6 times (Aalpha/beta-fibre units). The mean values of these variables in unresponsive units were similar to those of nociceptive units in each CV range; in C- and Adelta-fibre groups their mean AHP duration was even longer than in nociceptive units. 3. A-fibre LTM neurones were divided into Adelta- (D hair units, n = 8), and Aalpha/beta- (G hair/field units, n = 22; T (tylotrich) hair units, n = 6; rapidly adapting (RA) glabrous units, n = 6; slowly adapting (SA) hairy and glabrous units, n = 2; and muscle spindle (MS) units n = 17). MS and SA units had the shortest duration APs, FTs and AHP80s of all these groups. The mean RT in D hair units was significantly longer than in all Aalpha/beta LTM units combined. T hair units had the longest mean FT and AHP of all the A-LTM groups. The mean AHP was about 10 times longer in T hair units than in all other A-LTM units combined (significant), and was similar to that of A-fibre nociceptive neurones. 4. These differences in somatic AP shape may aid in distinguishing between LTM and nociceptive or unresponsive C- and Adelta-fibre units but probably not between nociceptive and unresponsive units. The differences seen may reflect differences in expression or activation of different types of ion channel.

Altered receptive fields and sensory modalities of rat VPL thalamic neurons during spinal strychnine-induced allodynia.

Altered receptive fields and sensory modalities of rat VPL thalamic neurons during spinal strychnine-induced allodynia. J. Neurophysiol. 78: 2296-2308, 1997. Allodynia is an unpleasant sequela of neural injury or neuropathy that is characterized by the inappropriate perception of light tactile stimuli as pain. This condition may be modeled experimentally in animals by the intrathecal (i.t.) administration of strychnine, a glycine receptor antagonist. Thus after i.t. strychnine, otherwise innocuous tactile stimuli evoke behavioral and autonomic responses that normally are elicited only by noxious stimuli. The current study was undertaken to determine how i.t. strychnine alters the spinal processing of somatosensory input by examining the responses of neurons in the ventroposterolateral thalamic nucleus. Extracellular, single-unit recordings were conducted in the lateral thalamus of 19 urethan-anaesthetized, male, Wistar rats (342 +/- 44 g; mean +/- SD). Receptive fields and responses to noxious and innocuous cutaneous stimuli were determined for 19 units (1 per animal) before and immediately after i.t. strychnine (40 microgram). Eighteen of the animals developed allodynia as evidenced by the ability of otherwise innocuous brush or air jet stimuli to evoke cardiovascular and/or motor reflexes. All (3) of the nociceptive-specific units became responsive to brush stimulation after i.t. strychnine, and one became sensitive to brushing over an expanded receptive field. Expansion of the receptive field, as determined by brush stimulation, also was exhibited by all of the low-threshold mechanoreceptive units (14) and wide dynamic range units (2) after i.t. strychnine. The use of air jet stimuli at fixed cutaneous sites also provided evidence of receptive field expansion, because significant unit responses to air jet developed at 13 cutaneous sites (on 7 animals) where an identical stimulus was ineffective in evoking a unit response before i.t. strychnine. However, the magnitude of the unit response to cutaneous air jet stimulation was not changed at sites that already had been sensitive to this stimulus before i.t. strychnine. The onset of allodynia corresponded with the onset of the altered unit responses (i.e., lowered threshold/receptive field expansion) for the majority of animals (9), but the altered unit response either terminated concurrently with symptoms of allodynia (6) or, more frequently, outlasted the symptoms of allodynia (10) as the effects of strychnine declined. The present results demonstrate that the direct, receptor-mediated actions of strychnine on the spinal processing of sensory information are reflected by changes in the receptive fields and response properties of nociceptive and nonnociceptive thalamic neurons. These changes are consistent with the involvement of thalamocortical mechanisms in the expression of strychnine-induced allodynia and, moreover, suggest that i.t. strychnine also produces changes in innocuous tactile sensation.

Functional evaluation of regenerated and misrouted axons to glabrous and hairy skin of the rat hind foot after sciatic neurotomy and suture

The function of misrouted regenerated polymodal nociceptor C-fibers and low-threshold mechanoreceptive axons in the lateral plantar nerve (LPN) and in the foot branch of the superficial peroneal nerve (fSPN) was evaluated 3 months after unilateral sciatic neurotomy and suture. Two weeks before evaluation the tibial fascicle (or the peroneal fascicle) above the neurotomy was cut and tied off. In this way only functional regeneration of misrouted axons was tested in the LPN (or the fSPN). In regenerated animals the glabrous skin area had no functional fSPN-related low-threshold mechanoreceptive axons. However, the hairy fSPN skin area showed function of misrouted LPN-related low-threshold mechanoreceptive axons. In both the glabrous skin domain innervated by the LPN and the hairy skin area supplied by the fSPN, functional regeneration of misrouted polymodal nociceptor C-fibers was found. We conclude that functional regeneration of misrouted axons related to polymodal nociceptive units and low-threshold mechanoreceptive units is more efficient in hairy skin of the rat foot whereas only misrouted polymodal nociceptor C-fibers recover function in glabrous skin.

Corneal Representation within the Trigeminal Subnucleus Caudalis and Adjacent Bulbar Lateral Reticular Formation of the Cat.

Corneal units in the trigeminal subnucleus caudalis and adjacent bulbar lateral reticular formation were studied in urethane-chloralose anesthetized cats. Corneal units were categorized into four classes: low-threshold corneal (LTC) units, high-threshold corneal (HTC) units, wide dynamic range (WDR) units with corneal input, and subnucleus reticularis ventralis (SRV) units with corneal input. Corneal receptive fields of these four classes of corneal afferent units consisted of 3-6 spots. Mechanical thresholds of LTC units were lower than 30 mg (2.6 g/mm2) and were comparable to the sensory threshold of the human cornea measured in patients with cataract. Mechanical thresholds of the other 3 classes of corneal afferent units were well above the pain threshold in the human cornea. LTC units were located in the magnocellular layer of trigeminal subnucleus caudalis and were intermingled with cutaneous low-threshold mechanoreceptive units. HTC units were coexistent with nociceptive specific units in the marginal layer and in the outer zone of substantia gelatinosa. WDR units with corneal input were found in the lateral part of trigeminal lamina V equivalent, which corresponds to the lateral part of subnucleus reticularis dorsalis. These 3 classes of corneal units were found at a level 2.7-3.5 mm caudal to the obex. SRV units were found in the dorsolateral part of SRV along the entire length of the medulla oblongata caudal to the obex. These results support the suggestion that either nonpainful sensation or pain can be evoked from the cornea.

Iontophoretic application of calcitonin gene-related peptide produces a slow and prolonged excitation of neurons in the cat lumbar dorsal horn

Calcitonin gene-related peptide (CGRP) was applied by iontophoresis onto physiologically characterized neurons. CGRP (20–100 Na) activated both wide-dynamic-range ( 5 8 ) and low-threshold mechanoreceptive units ( 3 12 ), but had no effect on nociceptive-specific neurons ( 0 4 ). The excitation was of slow onset (30 s to 3 min) and prolonged duration (up to 10 min). In none of the tested units did CGRP cause inhibition. The slow and prolonged action suggests a neuromodulatory role for CGRP in spinal cord sensory processes.

Cardiac sympathetic afferent input onto neurons in nucleus ventralis posterolateralis in cat thalamus

Neurons receiving cardiac sympathetic afferent input were studied in the nucleus ventralis posterolateralis (VPL) of the cat thalamus. Animals were anesthetized with urethan-chloralose. Units in the VPL were classified into 3 classes; low-threshold mechanoreceptive (LTM), nociceptive specific (NS) and wide dynamic range (WRD) units. Units driven by electrical stimulation of the left inferior cardiac nerve (ICN) were not included in the population of LTM units, but 43.5% of NS units and 68.8% of WDR units were excited by this stimulation. Units exclusively responsive to caridiac sympathetic afferents were found. Both NS and WDR units were located in the shell region of the caudal VPL. NS units responsive to cardiac afferents had a circumscribed cutaneous receptive field in the area corresponding to tactile dermatomes C 5–T 13. WDR units receiving cardiac sympathetic afferent input had at least a part of their receptive fields in the same area. These results thatthe shell region of the caudal VPL constitutes a thalamic link in a cardiac pain pathway, and that cardiac and cutaneous pain systems share a common projection locus in the VPL.

Thresholds of mechanosensitive afferents in the human hand as measured with von Frey hairs

Afferent activity in low threshold mechanoreceptive units innervating the glabrous skin area of the hand was recorded from the median nerve of human subjects, using tungsten needle electrodes. The units were classified as RA, PC, SA I and SA II units mainly on the basis of their adaptation and receptive field properties Thresholds of the units were determined by two methods, the force threshold was determined with von Frey hairs and the indentation threshold with triangular indentations of controlled amplitudes. The characteristics of the force developed by the von Frey hairs as a function of movement parameters was analyzed when the hairs were prodded against a force transducer. The rise time was 10–20 msec and the variability of the peak force was within-20% of the steady-state force during repetitive prodding with various repetition rates. The von Frey thresholds were determined for 284 units and the indentation threshold for 116 of them. The RA and PC units were the most sensitive unit types with median thresholds of 0.58 mN and 0.54 mN. The medians of the slowly adapting SA I and SA II units, were 1.3 mN and 7.5 mN. The threshold distributions for all unit types were positively skewed. No regional differences were found but the thresholds were identical in skin areas where the psychophysical thresholds have been shown to be diverse. The relation between force threshold and indentation threshold was analyzed for 116 units. There was a positive correlation between the two thresholds, although the relation appeared not to be identical for the four types of units and the scatter was considerable.

Tactile sensibility in the human hand: relative and absolute densities of four types of mechanoreceptive units in glabrous skin.

1. Single unit impulses were recorded with percutaneously inserted tungsten needle electrodes from the median nerve in conscious human subjects. 2. A sample of 334 low threshold mechanoreceptive units innervating the glabrous skin area of the hand were studied. In accordance with earlier investigations, the units were separated into four groups on the basis of their adaptation and receptive field properties: RA, PC, SA I and SA II units. 3. The locations of the receptive fields of individual units were determined and the relative unit densities within various skin regions were calculated. The over-all density was found to increase in the proximo-distal direction. There was a slight increase from the palm to the main part of the finger and an abrupt increase from the main part of the finger to the finger tip. The relative densities in these three regions were 1, 1.6, 4.2. 4. The differences in over-all density were essentially accounted for by the two types of units characterized by small and well defined receptive fields, the RA and SA I units, whereas the PC and SA II units were almost evenly distributed over the whole glabrous skin area. 5. The spatial distribution of densities supports the idea that the RA and SA I units account for spatial acuity in psychophysical tests. This capacity is known to increase in distal direction along the hand. 6. On the basis of histological data regarding the number of myelinated fibres in the median nerve, a model of the absolute unit density was proposed. It was estimated that the density of low threshold mechanoreceptive units at the finger tip is as high as 241 u./cm2, whereas in the palm it is only 58 u./cm2.