High-threshold Fibers Research Articles

Electrophysiologic research of surgically restored median nerve (experimental study).

In the experiment the possibility of providing effective regeneration of the injured median nerve in conditions of creation “protected” suture with intact ulnar nerve was studied. Experimental animals were divided into 4 groups: 1) complete neurotomy of the median nerve; 2) complete neurotomy of the median nerve with neurorrhaphy; 3) complete neurotomy of the median nerve and anastomosis creation between the median and the ulnar nerves in the distal site; 4) complete neurotomy of the median nerve and its neurorrhaphy with the anastomosis between the median and the ulnar nerves in the distal site. The histological study was carried out to identify the level of the nerve regeneration, state of the nerve distal segment and electrical conductivity of the surgically restored nerve on day 30 after the trauma. Morphometric and electrophysiological examinations showed that anastomosis creation allows to prevent atrophy and elimination of distal segment of median nerve and activate regenerative process in the injured nerve. The electrical conductivity of the nerve improved. An increased M-response of the forearm muscles was observed in groups with the anastomosis between the median and the ulnar nerves (the level of M-response recovery in the group 2 – 27,1%, group 3 – 18, 9%, group 4 – 84,6%). According to the study results we have made a conclusion that reinnervation of the muscles at an early stage of the median and the ulnar nerves regeneration was realized with the participation of high-threshold myelin nerve fibers.

Noise-induced and age-related hearing loss: new perspectives and potential therapies.

The classic view of sensorineural hearing loss has been that the primary damage targets are hair cells and that auditory nerve loss is typically secondary to hair cell degeneration. Recent work has challenged that view. In noise-induced hearing loss, exposures causing only reversible threshold shifts (and no hair cell loss) nevertheless cause permanent loss of >50% of the synaptic connections between hair cells and the auditory nerve. Similarly, in age-related hearing loss, degeneration of cochlear synapses precedes both hair cell loss and threshold elevation. This primary neural degeneration has remained a “hidden hearing loss” for two reasons: 1) the neuronal cell bodies survive for years despite loss of synaptic connection with hair cells, and 2) the degeneration is selective for auditory nerve fibers with high thresholds. Although not required for threshold detection when quiet, these high-threshold fibers are critical for hearing in noisy environments. Research suggests that primary neural degeneration is an important contributor to the perceptual handicap in sensorineural hearing loss, and it may be key to the generation of tinnitus and other associated perceptual anomalies. In cases where the hair cells survive, neurotrophin therapies can elicit neurite outgrowth from surviving auditory neurons and re-establishment of their peripheral synapses; thus, treatments may be on the horizon.

Modulation of afferent nerve activity by prostaglandin E2 upon urinary bladder distension in rats.

What is the central question of this study? It has been widely assumed that Cfibres innervating the bladder are mainly excited in overactive bladder syndrome. However, it remains unclear whether Aδ fibres are also activated in pathological conditions. What is the main finding and its importance? We found that a certain population of Aδ fibres, which become active specifically at a bladder pressure of more than 15cmH2 O in normal conditions, showed increased excitability in conditions of prostaglandin E2 -induced overactive bladder. This result suggests that a certain population of Aδ fibres, together with Cfibres, triggers pathophysiological activity. In overactive bladder syndrome, afferent Cfibres innervating the bladder show an increased activity level. However, it remains unclear whether all Cfibres are highly activated and whether Aδ fibres, the other type of bladder afferent fibre, are also involved in pathological conditions. To address these questions, we analysed the relationship between bladder pressure and single-unit firing patterns of afferent nerves in the left L6 dorsal roots in living rats. The recorded fibres were classified as Aδ fibres or Cfibres based on the response to 0.3μm tetrodotoxin. Certain populations of both Aδ fibres and Cfibres were activated at bladder pressures below 15cmH2 O (classified as low-threshold fibres), indicating their potential contribution to detection of normal bladder filling. Intravesical administration of prostaglandinE2 (PGE2 ) induced hyperexcitation in approximately half of such Cfibres, whereas the activity patterns of low-threshold Aδ fibres were unchanged. All fibres, regardless of type, which were almost silent in control conditions (classified as high-threshold fibres), were activated by application of PGE2 . Notably, the firing patterns of Aδ fibres, rather than Cfibres, were highly time locked to PGE2 -induced micro-oscillation of bladder pressure. These modulatory effects of PGE2 on Aδ fibres and Cfibres might trigger pathophysiological activity together in overactive bladder syndrome.

Purinergic Signalling in the Gut.

The article will begin with the discovery of purinergic inhibitory neuromuscular transmission in the 1960s/1970s, the proposal for purinergic cotransmission in 1976 and the recognition that sympathetic nerves release adenosine 5'-triphosphate (ATP), noradrenaline and neuropeptide Y, while non-adrenergic, non-cholinergic inhibitory nerve cotransmitters are ATP, nitric oxide and vasoactive intestinal polypeptide in variable proportions in different regions of the gut. Later, purinergic synaptic transmission in the myenteric and submucosal plexuses was established and purinergic receptors expressed by both glial and interstitial cells. The focus will then be on purinergic mechanosensory transduction involving release of ATP from mucosal epithelial cells during distension to activate P2X3 receptors on submucosal sensory nerve endings. The responses of low threshold fibres mediate enteric reflex activity via intrinsic sensory nerves, while high threshold fibres initiate pain via extrinsic sensory nerves. Finally, the involvement of purinergic signalling in an animal model of colitis will be presented, showing that during distension there is increased ATP release, increased P2X3 receptor expression on calcitonin gene-related peptide-labelled sensory neurons and increased sensory nerve activity.

Synaptopathy in the noise-exposed and aging cochlea: Primary neural degeneration in acquired sensorineural hearing loss

The classic view of sensorineural hearing loss (SNHL) is that the “primary” targets are hair cells, and that cochlear-nerve loss is “secondary” to hair cell degeneration. Our recent work in mouse and guinea pig has challenged that view. In noise-induced hearing loss, exposures causing only reversible threshold shifts (and no hair cell loss) nevertheless cause permanent loss of >50% of cochlear-nerve/hair-cell synapses. Similarly, in age-related hearing loss, degeneration of cochlear synapses precedes both hair cell loss and threshold elevation. This primary neural degeneration has remained hidden for three reasons: 1) the spiral ganglion cells, the cochlear neural elements commonly assessed in studies of SNHL, survive for years despite loss of synaptic connection with hair cells, 2) the synaptic terminals of cochlear nerve fibers are unmyelinated and difficult to see in the light microscope, and 3) the degeneration is selective for cochlear-nerve fibers with high thresholds. Although not required for threshold detection in quiet (e.g. threshold audiometry or auditory brainstem response threshold), these high-threshold fibers are critical for hearing in noisy environments. Our research suggests that 1) primary neural degeneration is an important contributor to the perceptual handicap in SNHL, and 2) in cases where the hair cells survive, neurotrophin therapies can elicit neurite outgrowth from spiral ganglion neurons and re-establishment of their peripheral synapses.This article is part of a Special Issue entitled <Auditory Synaptology>.

Hot Topics—Hidden hearing loss: Permanent cochlear-nerve degeneration after temporary noise-induced threshold shift

The classic view of sensorineural hearing loss (SNHL) is that the “primary” targets are hair cells, and that cochlear-nerve loss is “secondary” to hair cell degeneration. Our work in mouse and guinea pig has challenged that view. In noise-induced hearing loss, exposures causing only reversible threshold shifts (and no hair cell loss) nevertheless cause permanent loss of &amp;gt;50% of cochlear-nerve/hair-cell synapses. Similarly, in age-related hearing loss, degeneration of cochlear synapses precedes both hair cell loss and threshold elevation. This primary neural degeneration has remained hidden for two reasons: (1) the spiral ganglion cells, the cochlear neural elements commonly assessed in studies of SNHL, survive for years despite loss of synaptic connection with hair cells, and (2) the degeneration is selective for cochlear-nerve fibers with high thresholds. Although not required for threshold detection in quiet (e.g., threshold audiometry, auditory brainstem response threshold), these high-threshold fibers are critical for hearing in noisy environments. Our research suggests that (1) primary neural degeneration is an important contributor to the perceptual handicap in SNHL, and (2) noise exposure guidelines should be re-evaluated, as they are based on the faulty premise that threshold audiograms are the most sensitive measures of noise damage.

Purinergic signalling in the urinary tract in health and disease.

Purinergic signalling is involved in a number of physiological and pathophysiological activities in the lower urinary tract. In the bladder of laboratory animals there is parasympathetic excitatory cotransmission with the purinergic and cholinergic components being approximately equal, acting via P2X1 and muscarinic receptors, respectively. Purinergic mechanosensory transduction occurs where ATP, released from urothelial cells during distension of bladder and ureter, acts on P2X3 and P2X2/3 receptors on suburothelial sensory nerves to initiate the voiding reflex, via low threshold fibres, and nociception, via high threshold fibres. In human bladder the purinergic component of parasympathetic cotransmission is less than 3%, but in pathological conditions, such as interstitial cystitis, obstructed and neuropathic bladder, the purinergic component is increased to 40%. Other pathological conditions of the bladder have been shown to involve purinoceptor-mediated activities, including multiple sclerosis, ischaemia, diabetes, cancer and bacterial infections. In the ureter, P2X7 receptors have been implicated in inflammation and fibrosis. Purinergic therapeutic strategies are being explored that hopefully will be developed and bring benefit and relief to many patients with urinary tract disorders.

Neurological and cellular regulation of visceral hypersensitivity induced by chronic stress and colonic inflammation in rats

The role of inflammation in inducing visceral hypersensitivity (VHS) in ulcerative colitis patients remains unknown. We tested the hypothesis that acute ulcerative colitis-like inflammation does not induce VHS. However, it sets up molecular conditions such that chronic stress following inflammation exaggerates single-unit afferent discharges to colorectal distension. We used dextran sodium sulfate (DSS) to induce ulcerative colitis-like inflammation and a 9-day heterotypic chronic stress protocol in rats. DSS upregulated Nav1.8 mRNA in colon-responsive dorsal root ganglion (DRG) neurons, TRPV1 in colonic muscularis externae (ME) and BDNF in spinal cord without affecting the spike frequency in spinal afferents or VMR to CRD. By contrast, chronic stress did not induce inflammation but it downregulated Kv1.1 and Kv1.4 mRNA in DRG neurons, and upregulated TRPA1 and nerve growth factor in ME, which mediated the increase of spike frequency and VMR to CRD. Chronic stress following inflammation exacerbated spike frequency in spinal afferent neurons. TRPA1 antagonist suppressed the sensitization of afferent neurons. DSS-inflammation did not affect the composition or excitation thresholds of low-threshold and high-threshold fibers. Chronic stress following inflammation increased the percent composition of high-threshold fibers and lowered the excitation threshold of both types of fibers. We conclude that not all types of inflammation induce VHS, whereas chronic stress induces VHS in the absence of inflammation.

Noise-induced cochlear neuropathy is selective for fibers with low spontaneous rates

Acoustic overexposure can cause a permanent loss of auditory nerve fibers without destroying cochlear sensory cells, despite complete recovery of cochlear thresholds (Kujawa and Liberman 2009), as measured by gross neural potentials such as the auditory brainstem response (ABR). To address this nominal paradox, we recorded responses from single auditory nerve fibers in guinea pigs exposed to this type of neuropathic noise (4- to 8-kHz octave band at 106 dB SPL for 2 h). Two weeks postexposure, ABR thresholds had recovered to normal, while suprathreshold ABR amplitudes were reduced. Both thresholds and amplitudes of distortion-product otoacoustic emissions fully recovered, suggesting recovery of hair cell function. Loss of up to 30% of auditory-nerve synapses on inner hair cells was confirmed by confocal analysis of the cochlear sensory epithelium immunostained for pre- and postsynaptic markers. In single fiber recordings, at 2 wk postexposure, frequency tuning, dynamic range, postonset adaptation, first-spike latency and its variance, and other basic properties of auditory nerve response were all completely normal in the remaining fibers. The only physiological abnormality was a change in population statistics suggesting a selective loss of fibers with low- and medium-spontaneous rates. Selective loss of these high-threshold fibers would explain how ABR thresholds can recover despite such significant noise-induced neuropathy. A selective loss of high-threshold fibers may contribute to the problems of hearing in noisy environments that characterize the aging auditory system.

Hyperpolarization-activated cyclic nucleotide-gated cation channel subtypes differentially modulate the excitability of murine small intestinal afferents

To assess the role of hyperpolarization-activated cyclic nucleotide-gated cation (HCN) channels in regulating the excitability of vagal and spinal gut afferents. The mechanosensory response of mesenteric afferent activity was measured in an ex vivo murine jejunum preparation. HCN channel activity was recorded through voltage and current clamp in acutely dissociated dorsal root ganglia (DRG) and nodose ganglia (NG) neurons retrogradely labeled from the small intestine through injection of a fluorescent marker (DiI). The isoforms of HCN channels expressed in DRG and NG neurons were examined by immunohistochemistry. Ramp distension of the small intestine evoked biphasic increases in the afferent nerve activity, reflecting the activation of low- and high-threshold fibers. HCN blocker CsCl (5 mmol/L) preferentially inhibited the responses of low-threshold fibers to distension and showed no significant effects on the high-threshold responses. The effect of CsCl was mimicked by the more selective HCN blocker ZD7288 (10 μmol/L). In 71.4% of DiI labeled DRG neurons (n = 20) and 90.9% of DiI labeled NG neurons (n = 10), an inward current (I(h) current) was evoked by hyperpolarization pulses which was fully eliminated by extracellular CsCl. In neurons expressing I(h) current, a typical "sag" was observed upon injection of hyperpolarizing current pulses in current-clamp recordings. CsCl abolished the sag entirely. In some DiI labeled DRG neurons, the I(h) current was potentiated by 8-Br-cAMP, which had no effect on the I(h) current of DiI labeled NG neurons. Immunohistochemistry revealed differential expression of HCN isoforms in vagal and spinal afferents, and HCN(2) and HCN(3) seemed to be the dominant isoform in DRG and NG, respectively. HCNs differentially regulate the excitability of vagal and spinal afferent of murine small intestine.

Therapeutic potential of purinergic signalling for diseases of the urinary tract

This review begins with background information about the discovery and conceptual steps contributing to our current knowledge of purinergic signalling. It then deals with several topics concerned with the physiology and pathophysiology of the lower urinary tract, including: the involvement in the voiding reflex of ATP released as a co-transmitter with acetylcholine from parasympathetic nerves supplying the bladder and ATP released from urothelial cells during bladder distension to initiate the voiding reflex via P2X₃ receptors on suburothelial low-threshold sensory nerve fibres; this latter mechanosensory transduction pathway is also involved via high-threshold fibres in the initiation of pain. Treatment of prostate and bladder cancer with ATP not only appears to be effective against the primary tumours, but also improves the systemic symptoms associated with advanced malignancy. There is dual control of the tone of blood vessels: constriction by ATP released as a co-transmitter from sympathetic nerves and vasodilatation via ATP released from endothelial cells during shear stress acting on endothelial P2 receptors to release nitric oxide. A purinergic hypothesis is discussed for the mechanism underlying acupuncture, widely used for the treatment of urinary disorders.

Purinergic contribution to small intestinal afferent hypersensitivity in a murine model of postinfectious bowel disease

Increased sensitivity of the afferent innervation of the gastrointestinal tract reportedly underlies symptoms of discomfort and pain in functional bowel disorders. The present investigation aimed to examine whether the purinergic P2X(2) and P2X(3) receptor subunits contribute to the mechanosensitivity of small intestinal afferents in normal mice and in a murine model of postinfectious gut dysfunction. Mesenteric afferent nerve activity was recorded in a mouse jejunum preparation maintained in vitro. As has been shown previously, ramp distension of the jejunal segment evoked biphasic afferent discharge, reflecting activation of low and high threshold fibres. The average pressure-afferent response curve in mice deficient in both P2X(2) and P2X(3) subunits (n = 14) was not significantly different from that of the wild-type control preparations (n = 13). Application of pyridoxal 5-phosphate 6-azophenyl-2 ,4-disulphonic acid (PPADS) (30 micromol L(-1)), a P2X and P2Y antagonist, or 2,4,6-trinitrophenol-adenosine 5'-triphosphate (10 micromol L(-1)), an antagonist selective for homomeric P2X(3) and heteromeric P2X(2/3) receptors, had no effect on the averaged pressure-afferent response curve in wild-type animals. In Trichinella spiralis-infected mice, the magnitude of mesenteric afferent responses to jejunal distension was greater at day 21 and day 56 postinfection compared with the sham control preparations demonstrating the development of afferent hypersensitivity. PPADS had no significant effect upon mechanically evoked afferent discharge rates in sham treated preparations (n = 5), but significantly inhibited afferent sensitivity to jejunal distension in preparations from mice at day 21 (n = 6) and day 56 (n = 7) postinfection. These results suggest that purinergic mechanisms play no role in mechanosensory transduction in the normal small intestine but contribute significantly to postinfectious mechano-hypersensitivity.

Low-Threshold Primary Afferent Drive onto GABAergic Interneurons in the Superficial Dorsal Horn of the Mouse

Inhibition in the spinal cord dorsal horn is crucial for maintaining separation of touch and pain modalities. Disruption of this inhibition results in allodynia, allowing low-threshold drive onto pain and temperature-sensitive projection neurons. This low-threshold (LT) excitatory pathway is normally under strong inhibition. We hypothesized that superficial dorsal horn inhibitory neurons, which would be ideally located to suppress LT drive onto projection neurons in a feedforward manner, are driven by LT input. In addition, because disinhibition-induced allodynia shares some features with the immature dorsal horn such as elevated sensitivity to LT input, we also questioned whether LT drive onto inhibitory neurons changes during postnatal maturation. To investigate these questions, slices were made at different ages from transgenic mice with enhanced green fluorescent protein expression in GABAergic neurons and whole-cell recordings were made from these fluorescent neurons. Evoked synaptic activity was measured in response to electrical stimulation of the dorsal root. We demonstrate that Abeta fibers activate a significant proportion of superficial dorsal horn GABAergic neurons. This occurs with similar excitatory synaptic drive throughout postnatal maturation, but with a greater prevalence at younger ages. These GABAergic neurons are well situated to contribute to suppressing LT activation of output projection neurons. In addition, the majority of these GABAergic neurons also had convergent input from high-threshold fibers, suggesting that this novel subclass of GABAergic neurons is important for gating innocuous as well as noxious information.

Bladder afferent sensitivity in wild‐type and TRPV1 knockout mice

Understanding bladder afferent pathways may reveal novel targets for therapy of lower urinary tract disorders such as overactive bladder syndrome and cystitis. Several potential candidate molecules have been postulated as playing a significant role in bladder function. One such candidate is the transient receptor potential vanilloid 1 (TRPV1) ion channel. Mice lacking the TRPV1 channel have altered micturition thresholds suggesting that TRPV1 channels may play a role in the detection of bladder filling. The aim of this study was therefore to investigate the role of TRPV1 receptors in controlling bladder afferent sensitivity in the mouse using pharmacological receptor blockade and genetic deletion of the channel. Multiunit afferent activity was recorded in vitro from bladder afferents taken from wild-type (TRPV+/+) mice and knockout (TRPV1-/-) mice. In wild-type preparations, ramp distension of the bladder to a maximal pressure of 40 mmHg produced a graded increase in afferent activity. Bath application of the TRPV1 antagonist capsazepine (10 mum) caused a significant attenuation of afferent discharge in TRPV1+/+ mice. Afferent responses to distension were significantly attenuated in TRPV1-/- mice in which sensitivity to intravesical hydrochloric acid (50 mm) and capsaicin (10 microm) were also blunted. Altered mechanosensitivity occurred in the absence of any changes in the pressure-volume relationship during filling indicating that this was not secondary to a change in bladder compliance. Single-unit analysis was used to classify individual afferents into low-threshold and high-threshold fibres. Low threshold afferent responses were attenuated in TRPV1-/- mice compared to the TRPV1+/+ littermates while surprisingly high threshold afferent sensitivity was unchanged. While TRPV1 channels are not considered to be mechanically gated, the present study demonstrates a clear role for TRPV1 in the excitability of particularly low threshold bladder afferents. This suggests that TRPV1 may play an important role in normal bladder function.

Fatigue-induced modulation of the H reflex of soleus muscle in humans

We studied the effect of fatigue of the mm. gastrocnemius-soleus on the H reflex elicited by transcutaneous stimulation of n. tibialis and recorded from the m. soleus; healthy 18-to 34-year-old volunteers were tested. Fatigue was evoked by long-lasting (6 to 9 min) voluntary tonic static sole flexion of the foot (ankle extension) with a force equal to 75% of the maximum voluntary contraction (MVC). The amplitude of H reflex significantly (P < 0.001) decreased to about 60% of the initial value immediately after the period of fatiguing effort. Within 2 to 3 min, it relatively rapidly recovered and reached about 90% of the control, and this was followed by a period of slow recovery to about 96–97% of the initial value 30 min after conditioning fatigue. We suppose that the initial period of suppression of the H reflex results to a considerable extent in an increase in the intensity of presynaptic inhibition of transmission from Ia afferents due to tonic activation of high-threshold (groups III and IV) afferent fibers induced by intensive fatigue-related metabolic changes in the muscles. More long-lasting (tens of minutes) changes are related to slow reverse development of direct effects of fatigue-induced biochemical shifts in the muscle.

Purinergic mechanisms contribute to mechanosensory transduction in the rat colorectum

Background & Aims: Adenosine 5′-triphosphate plays a role in peripheral sensory mechanisms and, in particular, mechanosensory transduction in the urinary system. P2X 3 receptors are selectively expressed on small-diameter sensory neurons in the dorsal root ganglia; sensory neurons from dorsal root ganglia L1 and S1 supply the colorectum. This study investigated whether purinergic signaling contributes to mechanosensory transduction in the rat colorectum. Methods: A novel in vitro rat colorectal preparation was used to elucidate whether adenosine 5′-triphosphate is released from the mucosa in response to distention and to evaluate whether it contributes to sensory nerve discharge during distention. Results: P2X 3 receptor immunostaining was present on subpopulations of neurons in L1 and S1 dorsal root ganglia, which supply the rat colorectum. Distention of the colorectum led to pressure-dependent increases in adenosine 5′-triphosphate release from colorectal epithelial cells and also evoked pelvic nerve excitation, which was mimicked by application of adenosine 5′-triphosphate and α,β-methylene adenosine 5′-triphosphate. The sensory nerve discharges evoked by distention were potentiated by α,β-methylene adenosine 5′-triphosphate and ARL-67156, an adenosine triphosphatase inhibitor, and were attenuated by the selective P2X 1, P2X 3, and P2X 2/3 antagonist 2′,3′- O-trinitrophenyl-adenosine 5′-triphosphate and by the nonselective P2 antagonists pyridoxyl 5-phosphate 6-azophenyl-2′,4′-disulfonic acid and suramin. Adenosine, after ectoenzymatic breakdown of adenosine 5′-triphosphate, seems to be involved in the longer-lasting distention-evoked sensory discharge. Single-fiber analysis showed that high-threshold fibers were particularly affected by α,β-methylene adenosine 5′-triphosphate, suggesting a correlation between purinergic activation and nociceptive stimuli. Conclusions: Adenosine 5′-triphosphate contributes to mechanosensory transduction in the rat colorectum, and this is probably associated with pain.

Activation and sensitisation of low and high threshold afferent fibres mediated by P2X receptors in the mouse urinary bladder.

It has been proposed that extracellular ATP may be involved in visceral mechanosensory transduction by activating ligand-gated ion channels (P2X receptors). In this study, we have investigated the effects of the P2X(3) agonist alpha,beta-methylene ATP (alpha,beta-meATP) and antagonist 2',3'-O-trinitrophenyl-ATP (TNP-ATP) on pelvic afferents innervating the urinary bladder using an in vitro mouse bladder-pelvic nerve preparation. Intravesical application of alpha,beta-meATP (0.03-1 mM) increased multifibre discharges in a concentration-dependent manner. The agonist potentiated, whereas TNP-ATP (0.03 mM) attenuated, the multifibre responses to bladder distensions. Single-unit analysis revealed that both high threshold (HT) fibres (> 15 mmHg; known to be associated with nociception) and low threshold (LT) fibres (< 15 mmHg; probably associated with non-nociceptive events) could be induced to discharge by intravesical alpha,beta-meATP (1 mM, 0.1 ml). The response of the vast majority (21/22, 95.5 %) of HT fibres to bladder distensions was enhanced with a significantly reduced threshold and an increased peak response after exposure to the agonist. On the other hand, 59.7 % (46/77) of LT fibres showed a greater peak and a slightly reduced threshold for response to bladder distension in the presence of alpha,beta-meATP. An additional 11 'silent' fibres became mechanosensitive after exposure to alpha,beta-meATP. TNP-ATP (0.03 mM) did not affect the threshold of LT fibres, but it reduced the peak response of some (22/51, 43.1 %) LT fibres. Conversely, the antagonist resulted in a markedly elevated threshold and reduced peak activity in the majority (13/16, 81.3 %) of HT fibres. The results support the view that P2X(3) receptor-mediated mechanisms contribute to both nociceptive and non-nociceptive (physiological) mechanosensory transduction in the urinary bladder.

Cannabinoid-Induced Presynaptic Inhibition of Glutamatergic EPSCs in Substantia Gelatinosa Neurons of the Rat Spinal Cord

The effect of cannabinoids on excitatory transmission in the substantia gelatinosa was investigated using intracellular recording from visually identified neurons in a transverse slice preparation of the juvenile rat spinal cord. In the presence of strychnine and bicuculline, perfusion of the cannabinoid receptor agonist WIN55,212-2 reduced the frequency and the amplitude of spontaneous excitatory postsynaptic currents (sEPSCs). Furthermore, the frequency of miniature EPSCs (mEPSCs) was also decreased by WIN55,212-2, whereas their amplitude was not affected. Similar effects were reproduced using the endogenous cannabinoid ligand anandamide. The effects of both agonists were blocked by the selective CB(1) receptor antagonist SR141716A. Electrical stimulation of high-threshold fibers in the dorsal root evoked a monosynaptic EPSC in lamina II neurons. In the presence of WIN55,212-2, the amplitude of the evoked EPSC (eEPSCs) was reduced, and the paired-pulse ratio was increased. The reduction of the eEPSC following CB(1) receptor activation was unlikely to have a postsynaptic origin because the response to AMPA, in the presence of 1 microM TTX, was unchanged. To investigate the specificity of this synaptic inhibition, we selectively activated the nociceptive C fibers with capsaicin, which induced a strong increase in the frequency of EPSCs. In the presence of WIN55,212-2, the response to capsaicin was diminished. In conclusion, these results strongly suggest a presynaptic location for CB(1) receptors whose activation results in inhibition of glutamate release in the spinal dorsal horn. The strong inhibitory effect of cannabinoids on C fibers may thereby contribute to the modulation of the spinal excitatory transmission, thus producing analgesia at the spinal level.

Analysis of the Central Pathway of the Trigeminal-Salivary Reflex by the Spinal Trigeminal Tractotomy.

The aim of this study was to clarify the role of spinal trigeminal subnucleus interporalis (Sp 5 I) and subnucleus caudalis (Sp 5 C) on the trigeminal-salivary reflex. Extracellular recordings were made from a total of 119 superior salivatory nucleus (SSN) neurons innervating the submandibular and sublingual glands in urethane-chloralose anaesthetized cats. Responses to stimulation of the inferior alveolar (IA) nerve were investigated in trigeminal tractotomized animals at the level of the obex (Sp 5 C animals) and at the level of 2.5-3mm rostral from the obex (Sp 5 I animals) as well as in non-tractotomized (control) animals. In control, Sp 5 C and Sp 5 I animals, 50% (23/46), 46% (15/33) and 10% (4/40) of the identified SSN neurons orthodromically responded to stimulation of the IA nerve, respectively.In Sp 5 C animals, no significant reduction of the response rate was found in comparison with control animals, and there was no significant difference in the mean onset latency between control and Sp 5 C animals. However, only the responses activated by the medium-threshold afferent fibres or by high-threshold afferent fibres could be observed.In Sp 5 I animals, a marked reduction of the response rate was noted in comparison with control animals, and there was no significant difference in the mean onset latency between control and Sp 5 I animals. However, only the responses excited by the medium-threshold afferent fibres or by high-threshold afferent fibres could be observed.As the relay nucleus in the trigeminal-salivary reflex, Sp 5 I has a more important role and Sp 5 C has a lesser role.

Lignocaine selectively reduces C fibre-evoked neuronal activity in rat spinal cord in vitro by decreasing N-methyl- d-aspartate and neurokinin receptor-mediated post-synaptic depolarizations; implications for the development of novel centrally acting analgesics

The action of Lignocaine on nociceptive transmission in the spinal cord has been studied in vitro using ventral root potential (VRP) recordings from 10–12-day-old rat hemisected spinal cord preparations. Single-shock stimulation of a dorsal root at intensities sufficient to activate high-threshold C-primary afferent fibres elicited VRPs lasting for 15–20 sec in the corresponding ventral root. The VRP consisted of 3 distinct parts: the early, slow and prolonged components, as previously described (Thompson et al. 1992), where the early represents Aβ fibre-evoked mono- and polysynaptic responses lasting for tens of milliseconds, the slow is a largely N-methyl- d-aspartic acid (NMDA) receptor-mediated small-calibre afferent-generated component, lasting for about 1.5 sec, and the prolonged is a neurokinin receptor-mediated long-lasting component generated by high-threshold fibres. Lignocaine superfusion (40–60 μM) significantly and reversibly reduced the slow and prolonged components of the C fibre-evoked VRP in a dose-dependent manner without any effect on the early or Aβ fibre-mediated component of the VRP. The amplitude of the cumulative VRP generated by repetitive inputs (1 and 10 Hz) was also significantly reduced as was the depolarization produced by bath application of NMDA (100 μM) or substance P (SP, 1 μM) in the presence or absence of tetrodotoxin (TTX) (300 nM). At this dose range Lignocaine had no effect on the compound action potential (CAP) elicited by stimulating the sciatic nerve and recorded on the dorsal root. The CAP was only significantly reduced with a 300 μM) dose of Lignocaine. Application of the opiate, glycine, GABA A and GABA B receptor antagonists, naloxone (1 μM) strychnine (100 μM), bicuculline (100 μM) and phaclofen (100 μM) did not alter depressant effects of lignocaine on the VRP. Low concentrations of Lignocaine have a selective action on nociceptive transmission in the spinal cord which is different and more potent than its local anaesthetic conduction blockade in the periphery. This includes a reduction of direct or synaptically driven NMDA- and NK receptor-mediated post-synaptic depolarizations indicating that this class of sodium channel blockers may be potentially useful as analgesic agents, possibly acting on TTX-resistant sodium ion channels.