Low-threshold Mechanoreceptors Research Articles

Changes in somatic action potential shape in guinea-pig nociceptive primary afferent neurones during inflammation in vivo.

We have examined whether there are changes during inflammation in the membrane properties of nociceptive primary afferent neurones in the guinea-pig that might contribute to hyperalgesia. Inflammation was induced by intradermal injections of complete Freund's adjuvant (CFA) in the left leg. Intracellular voltage recordings were made from the somata of ipsilateral L6 and S1 dorsal root ganglion neurones in anaesthetised untreated guinea-pigs at 2 or 4 days after CFA treatment. 2. Units were classified as C, Adelta or Aalpha/beta on the basis of their dorsal root conduction velocities (CVs). Units with receptive fields on the left leg were characterized as nociceptive, low- threshold mechanoreceptive (LTM) or unresponsive according to their responses to mechanical and thermal stimuli. The shapes of their somatic action potentials (APs) evoked by dorsal root stimulation were recorded. 3. Comparisons of data from nociceptive neurones recorded in CFA treated animals after 2 and 4 days with data from CFA untreated (control) animals showed the following significant changes: in C-fibre nociceptors, decreased AP duration at base, AP rise time and AP fall time, and increased maximum rates of AP rise and fall with no change in afterhyperpolarization measured to 80 % recovery (AHP80); in Adelta-fibre nociceptors, decreased AP duration at base, AP fall time and a reduction in AHP80; and in Aalpha/beta-fibre nociceptors, a decreased AHP80 but no change in AP duration. Apart from a more negative membrane potential and AHP depth below 0 mV in Aalpha/beta nociceptors at 4 days compared with 2 days post-CFA, none of the above variables differed significantly between units recorded 2 or 4 days after CFA. Therefore the two groups were pooled and called CFA2 + 4d. 4. The reduction in AP duration in C-fibre nociceptors was apparent both in high threshold mechanoreceptor and polymodal nociceptors and also in units with either cutaneous or subcutaneous receptive fields. 5. No significant changes in AP duration at base or AHP80 were seen 2 or 4 days after CFA compared with control in either LTM or unresponsive neurones, although some of the latter may have become classified as nociceptors after CFA treatment. 6. The alterations in membrane properties of nociceptors should permit higher discharge frequencies, thus contributing to inflammatory hyperalgesia. They suggest active changes in the expression or activation of cation channels during peripheral inflammation.

Osteocalcin-immunoreactive primary sensory neurons in the rat spinal and trigeminal nervous systems

Osteocalcin-immunoreactivity (OC-ir) was examined in spinal and trigeminal primary sensory neurons of the adult rat. Sixteen percent of dorsal root ganglion (DRG) neurons were immunoreactive (ir) for this protein. These neurons were mostly large and measured 594–4583 μm 2 (mean±S.D.=2243±748 μm 2). Thirty-four percent of DRG neurons >1200 μm 2 and 4% of those in the range 600–1200 μm 2 showed the ir. Virtually all DRG neurons <600 μm 2 were devoid of OC-ir. In the trigeminal ganglion (TG), 25% of neurons exhibited the ir. Such neurons were of various sizes (range=156–2825 μm 2, mean±S.D.=1234±543 μm 2). Forty-five percent of TG neurons >800 μm 2 and 6% of those <400 μm 2 were immunoreactive for this protein. Twelve percent of TG neurons in the range 400–800 μm 2 showed the ir. In the mesencephalic trigeminal tract nucleus (Mes5), 63% of primary sensory neurons contained OC-ir. Virtually all OC-ir DRG and Mes5 neurons co-expressed parvalbumin-ir but not CGRP-ir. On the other hand, only 31% of OC-ir neurons co-expressed parvalbumin-ir and 10% co-expressed CGRP-ir in the TG. The present study indicates that DRG and Mes5 primary sensory neurons co-expressing OC- and parvalbumin-irs are spinal and trigeminal proprioceptors. OC-ir TG neurons which co-express parvalbumin- and CGRP-irs appear to include low-threshold mechanoreceptors and nociceptors, respectively.

An investigation of the neurophysiologic mechanisms of tourniquet-related pain: Changes in spontaneous activity and receptive field size in spinal dorsal horn neurons

Background and Objectives. Several theories have been proposed for the pain resulting from the nerve compression and ischemia associated with maintaining the inflation of a pneumatic tourniquet on an extremity. This investigation observes changes in the spontaneous activity and receptive field (RF) size of spinal dorsal horn neurons during tourniquet-related nerve compression and ischemia. Methods. Forty-eight pentobarbital-anesthetized rats were prepared for single-unit spinal dorsal horn cell recording utilizing an in vivo extracellular technique. Cells were characterized as either low threshold mechanoreceptor (LTM) neurons, wide dynamic range (WDR) neurons, or high threshold nociresponsive (NR) neurons based on their response to light touch (brush), non-noxious pressure, or noxious pinch. Receptive fields were carefully mapped. A 1 × 9 cm pneumatic tourniquet (Hokanson, Inc., Seattle, WA) was then applied to the animals' hind limb thigh and inflated to 300 mm Hg for 60 minutes. Throughout the 60-minute tourniquet inflation period, spontaneous activity, cell firing rate response characteristics, and RF size were carefully monitored. Results. Twenty-three cells in 20 animals were characterized and monitored: 8 LTM, 6 WDR, and 9 NR neurons. Receptive fields were distal to the tourniquet cuff for 16 cells and proximal for 7 cells. Low threshold mechanoreceptor neurons demonstrated little spontaneous activity prior to tourniquet inflation in the absence of RF stimulation. The RF size for all LTM neurons decreased or became silent during the tourniquet inflation period. The RF size increased in two of six WDR neurons with two cells demonstrating a progressive increase in intermittent spontaneous firing during the tourniquet inflation period. Receptive field size increased in five of five NR neurons with RF located proximal to the tourniquet cuff from 29 ± 10 mm 2 to 54 ± 30 mm 2 (RF area 1.9 ± 0.7; t = 2.7, P = .03). All five of these NR neurons also demonstrated an increase in spontaneous activity at 37 ± 14 minutes of tourniquet inflation which continued throughout the remainder of the tourniquet inflation period ( P < .05). Conclusions. The results of this study indicate that nerve compression and ischemia results in block of input to LTM neurons having RFs distal to the tourniquet cuff and an increase in spontaneous activity and expansion of the RFs of NRs, especially those with RFs located proximal to the tourniquet. Increases in spontaneous firing activity and expansion of the RFs of nociresponsive dorsal horn neurons receiving input from primary afferent nociceptors proximal to the tourniquet may explain, in part, the neurophysiologic mechanism of tourniquet-related pain.

Expression of alpha2-adrenergic receptors in rat primary afferent neurones after peripheral nerve injury or inflammation.

1. Immunocytochemistry with polyclonal antibodies directed against specific fragments of intracellular loops of alpha2A- and alpha2C-adrenergic receptors (alpha2A-AR, alpha2C-AR) was used to explore the possibility that expression of these receptors in dorsal root ganglion (DRG) neurones of rat alters as a result of peripheral nerve injury or localized inflammation. 2. Small numbers of neurones with positive alpha2A-AR immunoreactivity (alpha2A-AR-IR) were detected in DRG from normal animals or contralateral to nerve lesions. In contrast, after complete or partial sciatic nerve transection the numbers of ipsilateral L4 and L5 DRG somata expressing alpha2A-AR-IR sharply increased (>5-fold). There was no discernible change in the number of DRG neurones exhibiting alpha2A-AR-IR innervating a region in association with localized chemically induced inflammation. 3. After nerve injury, double labelling with Fluoro-Gold, a marker of retrograde transport from transected fibres, or by immunoreactivity for c-jun protein, an indicator of injury and regeneration, suggested that many of the neurones expressing alpha2A-AR-IR were uninjured by the sciatic lesions. 4. In general the largest proportionate increase in numbers of neurones labelled by alpha2A-AR-IR after nerve lesions appeared in the medium-large diameter range (31-40 microm), a group principally composed of cell bodies of low threshold mechanoreceptors. The number of small diameter DRG neurones labelled by alpha2A-AR-IR, a category likely to include somata of nociceptors, also increased but proportionately less. 5. Relatively few DRG neurones exhibited alpha2C-AR-IR; this population did not appear to change after either nerve lesions or inflammation. 6. These observations are considered in relation to effects of nerve injury on excitation of primary afferent neurones by sympathetic activity or adrenergic agents, sympathetically related neuropathy and reports of sprouting of sympathetic fibres in DRG.

C-fiber depletion alters response properties of neurons in trigeminal nucleus principalis.

The effects of C-fiber depletion induced by neonatal capsaicin treatment on the functional properties of vibrissa-sensitive low-threshold mechanoreceptive (LTM) neurons in the rat trigeminal nucleus principalis were examined in adult rats. Neonatal rats were injected either with capsaicin or its vehicle within 48 h of birth. The depletion of unmyelinated afferents was confirmed by the significant decrease in plasma extravasation of Evan's blue dye induced in the hindlimb skin of capsaicin-treated rats by cutaneous application of mustard oil and by the significant decrease of unmyelinated fibers in both the sciatic and infraorbital nerves. The mechanoreceptive field (RF) and response properties of 31 vibrissa-sensitive neurons in capsaicin-treated rats were compared with those of 32 vibrissa-sensitive neurons in control (untreated or vehicle-treated) rats. The use of electronically controlled mechanical stimuli allowed quantitative analysis of response properties of vibrissa-sensitive neurons; these included the number of center- and surround-RF vibrissae within the RF (i.e., those vibrissae which when stimulated elicited >/=1 and <1 action potential per stimulus, respectively), the response magnitude and latency, and the selectivity of responses to stimulation of vibrissae in different directions with emphasis on combining both the response magnitude and direction of vibrissal deflection in a vector analysis. Neonatal capsaicin treatment was associated with significant increases in the total number of vibrissae, in the number of center-RF vibrissae per neuronal RF, and in the percentage of vibrissa-sensitive neurons that also responded to stimulation of other types of orofacial tissues. Compared with control rats, capsaicin-treated rats showed significant increases in the response magnitude to stimulation of surround-RF vibrissae as well as in response latency variability to stimulation of both center- and surround-RF vibrissae. C-fiber depletion also significantly altered the directional selectivity of responses to stimulation of vibrissae. For neurons with multiple center-RF vibrissae, the proportion of center-RF vibrissae with net vector responses oriented toward the same quadrant was significantly less in capsaicin-treated compared with control rats. These changes in the functional properties of principalis vibrissa-sensitive neurons associated with marked depletion of C-fiber afferents are consistent with similarly induced alterations in LTM neurons studied at other levels of the rodent somatosensory system, and indeed may contribute to alterations previously described in the somatosensory cortex of adult rodents. Furthermore, these results provide additional support to the view that C fibers may have an important role in shaping the functional properties of LTM neurons in central somatosensory pathways.

The relationship between axonal spike shape and functional modality in cutaneous C-fibres in the pig and rat

Axonal spike shape was examined in identified cutaneous C-fibres dissected from the saphenous nerves of anaesthetized pigs and rats, and was found to vary with functional class. In the pig, the action potential duration for heat nociceptor units (duration at half peak amplitude, 1.25±0.16 ms, mean±S.E.M., n=32) was significantly longer than the duration for polymodal nociceptive units (0.88±0.11 ms, n=32). Both classes of nociceptive C-fibre had action potentials of longer duration than the low-threshold mechanoreceptor units (0.49±0.04 ms, n=24) and the inexcitable C-fibres (0.56±0.06 ms, n=19). Undershoot durations were also longer in nociceptive than non-nociceptive C-fibres. In contrast, spike amplitudes were similar in all classes of C-afferent. In the rat, as in the pig, the polymodal nociceptor units had action potentials of longer duration (0.75±0.05 ms, n=73) than the mechanoreceptor units (0.60±0.01 ms, n=23). C-fibres identified as spontaneously active sympathetic efferent units had wider action potentials (main initial peak: 1.01±0.12 ms, n=22; undershoot: 4.1±1.23 ms, n=20) than the afferent C-fibres (main peak: 0.69±0.03 ms, n=130; undershoot: 1.4±0.09 ms, n=111). All rat C-fibre types had action potentials with main initial peaks of a similar height. However, cold thermoreceptor units had spikes with significantly smaller undershoots compared to nociceptive or inexcitable C-fibres. It is concluded that there are clear differences in axonal spike shape between the different functional classes of C-fibre and, in particular, that nociceptive C-afferents tend to have axonal action potentials of longer duration than non-nociceptive afferents. However, there is too much overlap in the distributions of spike durations of different functional classes for this measure to be useful on its own for identifying the class of individual C-fibres. The ion channels responsible for the longer duration action potentials may provide a target for the development of highly selective analgesic drugs.

Activation of Pelvic Afferent Nerves from the Rat Bladder during Filling

This short paper has reviewed current information on the physiology of bladder mechanoreceptors. Afferents in the bladder mucosa appear to be sensitive to distension but not to contraction of the viscus, and are also sensitive to the chemical composition of the bladder contents. Evidence is provided that suggest that Neurokinin A is released from primary afferent endings in the submucosa and is involved in both the normal response to distension, and the sensitization process. The results suggest that the low threshold mechanoreceptors are polymodal, and are modulated by various mediators, including some peptides that may originate from the sensory endings themselves, and that these mediators may be active during normal filling.

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.

The distribution of primary afferent terminals from the eyelids of macaque monkeys.

Trigeminal sensory afferents from the eyelids convey two types of information that are important for the blink reflex. Movement of the lashes activates low-threshold mechanoreceptors which evoke protective blinks. Information about eyelid position is also transmitted centrally and is used to adapt the metrics of the blink reflex to changing conditions over time. This study employed transganglionic transport of horseradish peroxidase conjugated to choleratoxin-B subunit or wheat-germ agglutinin to investigate trigeminal afferents supplying the eyelids of macaque monkeys. Ganglion cells labeled from upper- and lower-lid injections were located in the ophthalmic and maxillary portions of the trigeminal ganglion, respectively. In both cases, labeled terminals were observed ipsilateral to the injected eyelid in the principal and spinal trigeminal nuclei. However, only a few labeled terminals were present in the principal nucleus, and very sparse terminal labeling was confined to a few locations along the ventral border of the pars oralis and interpolaris of the spinal trigeminal nucleus. The main concentration of label was found in the pars caudalis at and immediately below the spinomedullary junction. The terminal field from the upper eyelid was located ventrally in the pars caudalis, and that from the lower eyelid was located more dorsally. In both cases, the labeled terminal field was densest within lamina II of the spinal trigeminal nucleus. The heavy concentration of eyelid central terminals at the spinomedullary junction is surprising in light of physiological studies indicating representation of all parts of the face throughout the trigeminal nucleus. The distribution of eyelid afferent terminals in the macaque is caudal to the main concentration of corneal afferent terminals at the pars interpolaris/caudalis border. This may be a basis for differences seen in blinks produced by corneal as opposed to supraorbital stimulation. The presence of a single major site of eyelid primary afferent terminals suggests that sensory input for both eyelid proprioception and blink-reflex activation passes through this segment of the spinal trigeminal nucleus. These results provide a basis for investigation of the central connections of pars caudalis neurons in order to better establish the pathways producing trigeminally evoked blinks and blink adaptation.

Two spinal cords in birds: novel insights into early avian evolution

Restricted accessMoreSectionsView PDF ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmail Cite this article Woodbury C. Jeffery 1998Two spinal cords in birds: novel insights into early avian evolutionProc. R. Soc. Lond. B.2651721–1729http://doi.org/10.1098/rspb.1998.0494SectionRestricted accessTwo spinal cords in birds: novel insights into early avian evolution C. Jeffery Woodbury C. Jeffery Woodbury Department of Ornithology, American Museum of Natural History, 79th Street and Central ParkWest, NewYork, New York 10024, USA [email protected] Google Scholar Find this author on PubMed Search for more papers by this author C. Jeffery Woodbury C. Jeffery Woodbury Department of Ornithology, American Museum of Natural History, 79th Street and Central ParkWest, NewYork, New York 10024, USA [email protected] Google Scholar Find this author on PubMed Search for more papers by this author Published:22 September 1998https://doi.org/10.1098/rspb.1998.0494AbstractBirds can be subdivided into two large superordinal assemblages based on differences in the dorsal horn of the spinal grey matter. Palaeognaths (i.e. ratites and tinamous), along with a few other orders of neognathous birds, exhibit the primitive dorsal horn state characteristic of other amniotes wherein cutaneous nerves form a single map of the body surface across the dorsal horn. In contrast, the vast majority of neognaths exhibit a novel, distinctly bifid dorsal horn wherein cutaneous nerves form not one, but two separate maps of the skin, each lying side–by–side. This unusual dorsal horn organization, which has been highly conserved and represents the derived state in birds, may identify a novel, major avian clade. These findings shed new light on historically problematic taxa and the early evolutionary branching sequence among living birds. Most notably, they reveal that the traditional orders Gruiformes, Columbiformes, Cuculiformes and Piciformes are unnatural assemblages. Further, in addition to palaeognaths, these findings suggest that most gruiforms, including buttonquails and mesites, as well as pigeons, cuckoos, woodpeckers and songbirds, represent ancient lineages whose ancestry predates the majority of ‘modern’ birds. The phylogeny of living birds may thus be likened more to a dense bush than the traditional tree, with more than half of all living species arising from a basal side branch. Previous ArticleNext Article VIEW FULL TEXT DOWNLOAD PDF FiguresRelatedReferencesDetailsCited by Wild J (2015) The Avian Somatosensory System Sturkie's Avian Physiology, 10.1016/B978-0-12-407160-5.00005-1, (55-69), . Livezey B (2009) Phylogenetics of Modern Shorebirds (Charadriiformes) Based on phenotypic Evidence: I—Characterization, Bulletin of Carnegie Museum of Natural History, 10.2992/013.040.0101, 40, (1-96), Online publication date: 1-Dec-2009. Evrard H (2006) Estrogen synthesis in the spinal dorsal horn: a new central mechanism for the hormonal regulation of pain, American Journal of Physiology-Regulatory, Integrative and Comparative Physiology, 10.1152/ajpregu.00930.2005, 291:2, (R291-R299), Online publication date: 1-Aug-2006. LIVEZEY and ZUSI (2006) PHYLOGENY OF NEORNITHES, Bulletin of Carnegie Museum of Natural History, 10.2992/0145-9058(2006)37[1:PON]2.0.CO;2, 37, (1-544), Online publication date: 1-Jun-2006. Posso S and Donatelli R (2006) Análise filogenética e implicações sistemáticas e evolutivas nos Cuculiformes (Aves) com base na osteologia, comportamento e ecologia, Revista Brasileira de Zoologia, 10.1590/S0101-81752006000300003, 23:3, (608-629), Online publication date: 1-Sep-2006. Mayr G and Clarke J (2005) The deep divergences of neornithine birds: a phylogenetic analysis of morphological characters, Cladistics, 10.1111/j.1096-0031.2003.tb00387.x, 19:6, (527-553), Online publication date: 1-Dec-2003. Woodbury C, Ritter A and Koerber H (2001) Central anatomy of individual rapidly adapting low-threshold mechanoreceptors innervating the ?hairy? skin of newborn mice: Early maturation of hair follicle afferents, The Journal of Comparative Neurology, 10.1002/cne.1069, 436:3, (304-323), Online publication date: 30-Jul-2001. Woodbury C, Ritter A and Koerber H (2000) On the problem of lamination in the superficial dorsal horn of mammals: A reappraisal of the substantia gelatinosa in postnatal life, The Journal of Comparative Neurology, 10.1002/(SICI)1096-9861(20000131)417:1<88::AID-CNE7>3.0.CO;2-U, 417:1, (88-102), Online publication date: 31-Jan-2000. Homberger D and de Silva K (2000) Functional Microanatomy of the Feather-Bearing Integument: Implications for the Evolution of Birds and Avian Flight, American Zoologist, 10.1093/icb/40.4.553, 40:4, (553-574), Online publication date: 1-Aug-2000. Homberger D and de Silva K (2000) Functional Microanatomy of the Feather-Bearing Integument: Implications for the Evolution of Birds and Avian Flight1, American Zoologist, 10.1668/0003-1569(2000)040[0553:FMOTFB]2.0.CO;2, 40:4, (553-574), Online publication date: 1-Aug-2000. Tuinen M, Sibley C and Hedges S (2000) The Early History of Modern Birds Inferred from DNA Sequences of Nuclear and Mitochondrial Ribosomal Genes, Molecular Biology and Evolution, 10.1093/oxfordjournals.molbev.a026324, 17:3, (451-457), Online publication date: 1-Mar-2000. Dubbeldam J (2008) The Sensory Trigeminal System in Birds: Input, Organization and Effects of Peripheral Damage. A Review, Archives of Physiology and Biochemistry, 10.1076/apab.106.5.338.4367, 106:5, (338-345), Online publication date: 1-Jan-1998. This Issue22 September 1998Volume 265Issue 1407 Article InformationDOI:https://doi.org/10.1098/rspb.1998.0494Published by:Royal SocietyPrint ISSN:0962-8452Online ISSN:1471-2954History: Published online22/09/1998Published in print22/09/1998 License: Citations and impact Keywordsspinal cordsystematicsphylogenydorsal hornevolutionbirds Large datasets are available through Proceedings B's partnership with Dryad

Myelinated mechanically insensitive afferents from monkey hairy skin: heat-response properties.

To compare the heat responses of mechanically sensitive and mechanically insensitive A-fiber nociceptors, an electrical search technique was used to locate the receptive fields of 156 A-fibers that innervated the hairy skin in the anesthetized monkey (77 A beta-fibers, 79 A delta-fibers). Two-thirds of these afferents were either low-threshold mechanoreceptors (n = 91) or low-threshold cold receptors (n = 11). Nine A beta-fibers and 41 A delta-fibers were cutaneous nociceptors, and four A delta-fibers innervated subcutaneous tissue. The majority of cutaneous A-fiber nociceptors were heat sensitive (43/50 = 86%). Heat-insensitive cutaneous A-fiber nociceptors consisted of one cold nociceptor, three silent nociceptors, and three high-threshold mechanoreceptors. Two types of response were observed to an intense heat stimulus (53 degrees C, 30 s). Type I (n = 26) was characterized by a long latency (mean: 5 s) and a late peak discharge (16 s). Type II (n = 17) was characterized by a short latency (0.2 s) and an early peak discharge (0.5 s). Type I fibers exhibited faster conduction velocities (25 vs. 14 m/s) and higher heat thresholds (> 53 vs. 47 degrees C, 1-s duration) than type II fibers. The possibility that the type I heat response was a result of sensitization was tested in three fibers by determining the heat threshold to 30-s duration stimuli (42-46 degrees C). For this long stimulus duration heat thresholds were reproducible across multiple runs, and the threshold to the 1-s duration stimulus was not altered by these tests. Thus fibers with a type I heat response were not high-threshold mechanoreceptors that developed a heat response through sensitization. Fibers with a type II heat response had significantly higher mechanical thresholds (median: 15 bar) than fibers with a type I heat response (5 bar). This finding accounts for the observation that type II heat responses were infrequently observed in earlier studies wherein the search technique depended on mechanical responsiveness. Fibers with a type II response exhibited a graded response to heat stimuli, marked fatigue to repeated applications of heat stimuli, and adaptation to sustained heat stimuli similar to that seen in C-fiber nociceptors. First pain sensation to heat is served by type II A-fiber nociceptors that are mechanically insensitive. Type I A-fiber nociceptors likely signal pain to long-duration heat stimuli and may signal first pain sensation to mechanical stimuli.

Sensory changes in the territory of the lingual and inferior alveolar nerves following lower third molar extraction

Post-injury inflammation activates nociceptive systems and recruits normally non-nociceptive afferents into a pain processing role. During inflammation, A β low threshold mechanoreceptor afferents that usually mediate tactile sensation acquire properties of nociceptors, allowing them to participate in post-injury spontaneous pain and evoked abnormalities such as tenderness and pain to light touch. This study assessed the sensory consequences of post-injury inflammation following extraction of a single, lower third molar tooth. Extensive bilateral evaluations were performed in the territory of nerves assumed to be exposed to both inflammation and mechanical trauma, inflammation alone, or only the central consequences of peripheral inflammation. Testing at the distal termination of nerves assumed to be exposed to local inflammation (mental and lingual nerve territory) revealed decreased detection thresholds ( P<0.05) to electrical stimulation and to mechanical stimulation by sensitive, disposable filaments developed and validated for this application. Testing at sites of assumed inflammation and mechanical trauma (mental nerve territory) showed reduced pain thresholds to electrical stimulation. Thermal detection and pain thresholds were not altered at any location in patients, and no effects were observed in control subjects receiving only local anesthetic injections. These results in humans are consistent with recent experimental evidence that inflammatory processes alter the central consequence of activity in large-diameter A β touch primary afferents evoked under natural conditions by gentle mechanical stimulation. These effects result in hyperesthesia, increased sensitivity to light touch, and mechanical allodynia, pain evoked by normally innocuous stimulation of A β primary afferents.

Responses of dorsal column nuclei neurons in rats with experimental mononeuropathy

To examine the functional role of dorsal column nuclei in neuropathic pain, electrophysiological properties of low- and high-threshold dorsal column nuclei neurons in neuropathic and normal rats were examined. Single-neuronal activities were recorded from the gracile nucleus (GN) in rats at 10–14 days after application of four loose ligatures around the sciatic nerve (chronic constriction nerve injury; CCI). A total of 190 units were recorded from the GN in naive and CCI rats. The largest population of low-threshold mechanoreceptive (LTM) neurons recorded from the GN of naive rats were classified as rapidly-adapting (RA) LTM neurons, whereas those from CCI rats were slowly-adapting (SA) neurons. Mean orthodromic latencies of GN neurons ipsilateral to the CCI after sciatic nerve stimulation were significantly longer than those of naive animals and those of GN neurons without receptive fields were significantly longer than any other type of neurons. One hundred and eight of 190 GN neurons were also antidromically activated following electrical stimulation of the ventro-lateral posterior nucleus of the thalamus. Furthermore, when stronger stimuli were applied to the sciatic nerve, some GN neurons also responded with long latencies. GN neurons of sham-operated naive rats and those contralateral to the CCI had mechanical receptive fields on the paw, whereas 8.0% of the GN neurons in the CCI side did not have any detectable mechanical receptive field. Receptive field size was not significantly different between neurons ipsilateral or contralateral to the CCI and those of naive rats. Spontaneous activity of GN neurons from the ipsilateral side was significantly higher than those from the contralateral side. On the other hand, spontaneous activity of GN neurons both ipsilateral and contralateral to the CCI were significantly higher than those from naive rats. Furthermore, afterdischarges of GN neurons ipsilateral and contralateral to the CCI were significantly higher than those of naive rats. The present data suggest that the dorsal column pathway is involved in CCI-produced sensory abnormalities by conveying their hyperactivity to thalamic neurons.

Discharge patterns of intramural mechanoreceptive afferents during selective distension of the cat's rectum.

The afferent input from the rectum to the central nervous system (CNS) has yet to be thoroughly characterized. The characteristics of mechanoreceptive rectal afferents have been studied in unanaesthetized decerebrate cats. Following lumbo-sacral laminectomy, single-unit activity (occasionally multi-unit activity) was recorded from centrally cut filaments of the sacral dorsal roots (predominantly S2), while a balloon was inflated in the rectum. Starting from their background activities (mean 15.1 imp sec-1, SD 7.6 imp sec-1), afferent discharge rate increased with increasing balloon pressure (mean threshold 6.3 mmHg, SD 3.6 mmHg). The dependence of firing rate on intrarectal pressure began to flatten out at 25 mmHg (mean; SD 10 mmHg). For 22 out of 29 units (76%) complete saturation occurred at 35 mmHg (mean; SD 15 mmHg) with a maximum discharge rate of 31 imp sec-1 (mean; SD 12.6 imp sec-1). In a number of recording sessions, cyclical rectal contractions were observed. In these cases, changes in firing of the units were closely related to changes in intrarectal pressure. Pressure-related afferent activity could be enhanced by parasympathomimetic drugs which augmented rectal contractions. We conclude that sacral dorsal roots contain afferents from low-threshold mechanoreceptors located in the rectal wall, and that these afferents monitor the filling state and contraction level of the rectum.

Relationship of substance P to afferent characteristics of dorsal root ganglion neurones in guinea-pig.

1. The relationship between the afferent properties and substance P-like immunoreactivity (SP-LI) of L6 and S1 dorsal root ganglion (DRG) neuronal somata was examined in anaesthetized guinea-pigs. Glass pipette microelectrodes filled with fluorescent dyes were used to make intracellular recordings and to label DRG somata. The dorsal root conduction velocity (CV) and the afferent receptive properties of each unit were categorized according to criteria established in other species. Categories included a variety of low threshold mechanoreceptive classes, innocuous thermoreceptive and several nociceptive classes. Nociceptive units were further subdivided on the basis of CV and the locus of the receptive field (superficial cutaneous, deep cutaneous or subcutaneous). 2. SP-LI was determined using the avidin-biotin complex method and the relative staining intensity determined by image analysis. The possible significance of labelling intensity is discussed. Clear SP-LI appeared in twenty-nine of 117 dye-labelled neurones. All SP-LI positive units with identified receptive properties were nociceptive but not all categories of nociceptors were positive. The intensity of SP-LI labelling varied, often systematically, in relation to afferent properties. There was a tendency for nociceptive neurones with slower CVs and/or smaller cell bodies to show SP-LI. 3. Nineteen of fifty-one C fibre neurones showed SP-LI. Fewer than half the C polymodal nociceptors (CPMs) were positive. The most intensely labelled units were the deep cutaneous nociceptors and some of the CPMs in glabrous skin. C low threshold mechanoreceptors and cooling-sensitive units did not show SP-LI. 4. Ten of sixty-six A fibre neurones exhibited SP-LI, including eight of sixteen A delta nociceptors and two of fifteen A alpha/beta nociceptors. A fibre neurones exhibiting SP-LI included seven of eight deep cutaneous mechanical nociceptors and some superficial cutaneous mechano-heat nociceptors of hairy skin. In contrast, none of twenty superficial cutaneous A high threshold mechanoreceptor units or the thirty-five A fibre low threshold units (D-hair and other units) showed detectable SP-LI. 5. We conclude that SP-LI labelling in guinea-pig DRG neurones is related to (a) afferent receptive properties, (b) the tissue in which the peripheral receptive terminals are located, (c) the CV and (d) the soma size.

Responses of primary afferents and spinal dorsal horn neurons to thermal and mechanical stimuli before and during zymosan-induced inflammation of the rat hindpaw

Intraplantar administration of zymosan produces inflammation and results in behavioral evidence of hyperalgesia to mechanical and thermal stimuli in the rat. In the present studies, responses of primary afferents and spinal dorsal horn neurons to mechanical and thermal stimuli were examined before and during zymosan-induced inflammation of the hindpaw. In tests of responses of primary afferents to mechanical stimuli, group mean mechanical response thresholds of C-mechanonociceptor (CMN) units significantly decreased after zymosan administration. The group mean mechanical response thresholds of low threshold mechanoreceptor (LTM) units, A-mechanoheat (AMH) units, high threshold mechanoreceptor (HTM) units, and C-mechanoheat (CMH) units showed either no change or were increased significantly by intraplantar administration of zymosan. The group mean total discharges evoked during the 10 s mechanical stimulus were significantly increased after zymosan administration in CMN units. The group mean total discharges were either significantly decreased or unchanged in LTM, AMH, HTM, and CMH units. In tests of responses of spinal dorsal horn neurons to mechanical stimuli, the group mean mechanical response threshold of nociceptive specific (NS) units decreased significantly 1 h following administration of zymosan, whereas no significant changes occurred in the mechanical response thresholds of wide dynamic range (WDR) neurons in zymosan-injected rats, WDR neurons in saline-injected rats, or NS neurons in saline-injected rats. The group mean total discharges of only NS neurons were significantly increased during the 10 s mechanical stimulus 3 and 4 h after zymosan administration. In tests of responses of primary afferents to thermal stimuli, intraplantar administration of zymosan resulted in significant decreases in group mean response thresholds of CMH units and significant increases in group mean response thresholds of AMH units. The group mean total discharges of CMH units was either unchanged or significantly increased during thermal stimuli depending on both the time of testing and the temperature of the test stimulus. The group mean total number of discharges of AMH units was significantly decreased during tests of all thermal stimuli. In tests of responses of spinal dorsal horn neurons to thermal stimuli, intraplantar administration of zymosan resulted in significant decreases in thermal response thresholds of both WDR and NS units of zymosan-injected rats, but no changes in WDR and NS units of saline-injected rats. The group mean total discharges evoked by the 15 s thermal stimuli also increased significantly in both WDR and NS units after zymosan administration. Zymosan administration resulted in increased background activity only in CMH units. These increases occurred immediately following the injection and dissipated by the first hourly test period. Significant changes in background discharges of both WDR and NS units occurred at some hourly test intervals following administration of zymosan, but these changes were not consistent with respect to either unit type or modality of the test stimulus. These data suggest that the zymosan-induced hyperalgesia to mechanical stimuli observed in behavioral studies reflects decreases in response thresholds of peripheral CMN units and spinal NS neurons. Hyperalgesia to thermal stimuli reflects decreases in response thresholds of peripheral CMH units, spinal WDR neurons, and spinal NS neurons. These data support the view that different physiological substrates mediate hyperalgesia to either thermal or mechanical stimuli following intraplantar administration of zymosan.

Coexpression of calretinin and parvalbumin in Ruffini-like endings in the rat incisor periodontal ligament

The coexpression of calretinin- (CR) and parvalbumin-immunoreactivities (irs) was examined in oro-facial tissues of the rat. Nerve fibers coexpressing these calcium-binding proteins (CaBPs) were observed in the lingual periodontal ligament of incisors but not other tissues. In the part of periodontal ligament adjacent to the alveolar bone, such nerve fibers left nerve bundles and formed bush-like endings, i.e., they ramified repeatedly and terminated with one to four twigs. An immunoelectron microscopic method indicated that these endings were identical to Ruffini-like endings. 4% of trigeminal neurons retrogradely labeled from the inferior alveolar nerve coexpressed CR- and parvalbumin-irs. The present observations suggest that the coexpression of these CaBPs may be a specific marker for low-threshold mechanoreceptors in the trigeminal ganglion.

Differential effects of ketamine and MK-801 on A-fiber and C-fiber responses of spinal wide dynamic range neurons in the cat.

To clarify whether ketamine suppresses both A-fiber-and C-fiber-mediated pain and to compare the effects of ketamine with those of MK-801. Experiments were performed on urethane/chloralose-anesthetized cats. Glass capillary microelectrodes were used to record extracellular single unit activities from wide dynamic range (WDR) neurons in the spinal dorsal horn. Responses evoked by electrical stimulation of the superficial peroneal (SP), posterior tibial (PT), or both nerves were analyzed. The responses to successive electrical stimuli were displayed on a personal computer using a raster-dot processing program. A subanesthetic dose of intravenous ketamine suppressed both A- and C-fiber responses of WDR neurons in a dose-dependent manner without affecting A-fiber response of low-threshold mechanoreceptive (LTM) neurons. The C-fiber response was more markedly suppressed than the A-fiber response. Intravenous administration of MK-801, a specificN-methyl-D-aspartate (NMDA) antagonist, selectively suppressed the C-fiber response of WDR neurons. Intravenous ketamine may suppress both A-and C-fiber-mediated pain at a subanesthetic dose. This finding could be a scientific basis for the usefulness of ketamine during clinical procedures such as dressing changes or débridement of the burned patient.

Parvalbumin- and calbindin D-28k-immunoreactive innervation of orofacial tissues in the rat.

Parvalbumin- and calbindin D-28k-immunoreactive (-ir) innervation was examined in orofacial tissues of the rat. Labial and facial skins were devoid of the calcium-binding protein (CaBP)-ir nerve endings, while the infraorbital and mental nerves contained numerous parvalbumin-ir axons. Labial and gingival mucosae were also devoid of the CaBP-ir nerve endings. The buccal mucosa and incisive papilla contained both encapsulated and unencapsulated endings, while the hard palate mucosa excluding the incisive papilla contained only unencapsulated endings. Encapsulated endings were found just beneath the epithelium or attached to the cartilaginous core of the incisive papilla. Unencapsulated endings in the lamina propria were subdivided into two types: simple (unramified) and complex (ramified). Neurites of simple endings were straight, curved, or coiled, while those of complex endings exhibited a bush-like appearance due to the ramification. In addition, palatal rugae contained intraepithelial endings. The unencapsulated complex endings in palatal rugae coexpressed parvalbumin- and calbindin D-28k-irs, whereas other endings were immunoreactive for parvalbumin alone. The pterygopalatine ganglion contained calbindin D-28k-ir pericellular fibers but not the ir cell bodies. A subpopulation of trigeminal ganglion neurons coexpressed both CaBPs. CaBP-ir encapsulated and unencapsulated endings in the oral mucosa probably include low-threshold mechanoreceptors, while parvalbumin-ir intraepithelial endings in the palatal mucosa may be involved in nociception.

Responses of cutaneous A-fiber nociceptors to noxious cold.

Responses of cutaneous nociceptors to natural stimuli, particularly mechanical and heat stimuli, have been well documented. Although nociceptors are excited by noxious cold stimuli, there have been few studies of their stimulus-response functions for cold stimuli over a wide range of stimulus temperatures. Furthermore, the proportion of nociceptors excited by noxious cold is not clear. In the present study, we examined responses of mechanosensitive A delta-nociceptors and low-threshold mechanoreceptors to a wide range of cold stimuli that included stimulus temperatures <0 degrees C. Electrophysiological recordings were made from single primary afferent fibers in the saphenous nerves of anesthetized rats. Cutaneous sensory receptors were classed according to their conduction velocity and subgrouped functionally according to their responses evoked by mechanical, heat, and cold stimuli (0 degrees C). Responses evoked by a wide range of cold stimulus intensities that included stimuli considered innocuous and noxious (painful) were then assessed. Stimuli of 20 to -20 degrees C were delivered to the receptive field via a 1-cm2 contact thermode from a base temperature of 32 degrees C. Stimuli were applied in descending order of 2 degrees C decrements. Stimulus ramp rate was 5 degrees C/s, and stimulus temperatures were applied for a duration of 10 s. A total of 90 A fibers was studied, of which 61 were nociceptors and had conduction velocity in the A delta-range (2-30 m/s). Nociceptors were classed initially as mechanical, mechanoheat, and mechanocold nociceptors. The remaining 29 fibers were low-threshold mechanoreceptors with conduction velocity in the A delta- or A beta-range (>30 m/s). These were subgrouped according to their adaptive properties as slowly or rapidly adapting, and according to whether they were excited by hair movement (hair follicle afferent fibers). All nociceptors were excited by noxious cold. Only 30% of nociceptors were considered sensitive to cold on initial classification with the use of a cold stimulus of 0 degrees C. However, all nociceptors were excited by stimulus intensities <0 degreesC. Response thresholds for cold ranged from 14 to -18 degrees C (-4.6 +/- 1.07 degrees C, mean +/- SE). The total number of impulses, discharge rate, and peak discharge increased monotonically as intensity of cold stimuli increased. Power functions were used to determine the rate at which the number of impulses increased as stimulus intensity increased. The slopes of power funcions ranged from 0.12 to 2.28 (mean 1.07 +/- 0.13). Most mechanoreceptors were not excited by cold stimuli. The only types of mechanoreceptors that responded reliably to cold stimuli were the slowly adapting mechanoreceptors. Responses usually occurred during the temperature ramp when the skin temperature was decreasing. There was no evidence that mechanoreceptors encoded the intensity of cold stimuli at intensities above or below 0 degrees C, because evoked responses did not increase with intensity of cold stimuli. It is concluded that the proportion of cutaneous A delta-nociceptors excited by noxious cold stimuli has been underestimated in previous studies. All nociceptors were excited by stimulus temperatures <0 degrees C and encoded the intensity of cold stimuli. It is therefore likely that cutaneous A delta-nociceptors contribute to the sensation of cold pain, particularly pain produced by stimulus temperatures <0 degrees C.