Excitability Of Primary Sensory Neurons Research Articles

Emerging role of Toll-like receptors in the control of pain and itch

Toll-like receptors (TLRs) are germline-encoded pattern-recognition receptors that initiate innate immune responses by recognizing molecular structures shared by a wide range of pathogens, known as pathogen-associated molecular patterns (PAMPs). After tissue injury or cellular stress, TLRs also detect endogenous ligands known as danger-associated molecular patterns (DAMPs). TLRs are expressed in both non-neuronal and neuronal cell types in the central nervous system (CNS) and contribute to both infectious and non-infectious disorders in the CNS. Following tissue insult and nerve injury, TLRs (such as TLR2, TLR3, and TLR4) induce the activation of microglia and astrocytes and the production of the proinflammatory cytokines in the spinal cord, leading to the development and maintenance of inflammatory pain and neuropathic pain. In particular, primary sensory neurons, such as nociceptors, express TLRs (e.g., TLR4 and TLR7) to sense exogenous PAMPs and endogenous DAMPs released after tissue injury and cellular stress. These neuronal TLRs are new players in the processing of pain and itch by increasing the excitability of primary sensory neurons. Given the prevalence of chronic pain and itch and the suffering of affected people, insights into TLR signaling in the nervous system will open a new avenue for the management of clinical pain and itch.

Enhanced Excitability of Small Dorsal Root Ganglion Neurons in Rats with Bone Cancer Pain

BackgroundPrimary and metastatic cancers that affect bone are frequently associated with severe and intractable pain. The mechanisms underlying the development of bone cancer pain are largely unknown. The aim of this study was to determine whether enhanced excitability of primary sensory neurons contributed to peripheral sensitization and tumor-induced hyperalgesia during cancer condition. In this study, using techniques of whole-cell patch-clamp recording associated with immunofluorescent staining, single-cell reverse-transcriptase PCR and behavioral test, we investigated whether the intrinsic membrane properties and the excitability of small-sized dorsal root ganglion (DRG) neurons altered in a rat model of bone cancer pain, and whether suppression of DRG neurons activity inhibited the bone cancer-induced pain.ResultsOur present study showed that implantation of MRMT-1 tumor cells into the tibial canal in rats produced significant mechanical and thermal hyperalgesia in the ipsilateral hind paw. Moreover, implantation of tumor cells provoked spontaneous discharges and tonic excitatory discharges evoked by a depolarizing current pulse in small-sized DRG neurons. In line with these findings, alterations in intrinsic membrane properties that reflect the enhanced neuronal excitability were observed in small DRG neurons in bone cancer rats, of which including: 1) depolarized resting membrane potential (RMP); 2) decreased input resistance (Rin); 3) a marked reduction in current threshold (CT) and voltage threshold (TP) of action potential (AP); 4) a dramatic decrease in amplitude, overshot, and duration of evoked action potentials as well as in amplitude and duration of afterhyperpolarization (AHP); and 5) a significant increase in the firing frequency of evoked action potentials. Here, the decreased AP threshold and increased firing frequency of evoked action potentials implicate the occurrence of hyperexcitability in small-sized DRG neurons in bone cancer rats. In addiotion, immunofluorescent staining and single-cell reverse-transcriptase PCR revealed that in isolated small DRG neurons, most neurons were IB4-positive, or expressed TRPV1 or CGRP, indicating that most recorded small DRG neurons were nociceptive neurons. Finally, using in vivo behavioral test, we found that blockade of DRG neurons activity by TTX inhibited the tumor-evoked mechanical allodynia and thermal hyperalgesia in bone cancer rats, implicating that the enhanced excitability of primary sensory neurons underlied the development of bone cancer pain.ConclusionsOur present results suggest that implantation of tumor cells into the tibial canal in rats induces an enhanced excitability of small-sized DRG neurons that is probably as results of alterations in intrinsic electrogenic properties of these neurons. Therefore, alterations in intrinsic membrane properties associated with the hyperexcitability of primary sensory neurons likely contribute to the peripheral sensitization and tumor-induced hyperalgesia under cancer condition.

The modulation of the excitability of primary sensory neurons by Ca2+–CaM–CaMKII pathway

Ca(2+)-calmodulin (CaM) dependent protein kinase II (CaMKII) is an important intracellular signal transduction pathway. CaMKII is rich in the primary sensory neurons and specifically presents in the small- and medium-sized neurons. It remains unclear about the modulation on the excitability of primary sensory neurons by Ca(2+)-CaM-CaMKII pathway. By current clamp recording, we found that the excitability of capsaicin-sensitive small and medium trigeminal ganglion (TG) neurons was significantly reduced by a CaM specific antagonist (W-7) and a CaMKII antagonist (KN-93). The inhibition is represented as the reduction of numbers of action potential (AP), decrease of the amplitude of AP, increase of threshold, and prolongation of duration of AP. Consistently, by voltage clamp recording, we found that both voltage-gated sodium channels (VGSCs) and voltage-gated potassium channels (VGPCs) were inhibited by W-7 and KN-93 in the order of total sodium (Na(+)) current (INa-T) > sustained potassium (K(+)) current (IK) > A-type K(+) current (IA). In addition, AIP (a selective CaMKII peptide inhibitor) and KN-93 caused a similar inhibition of INa-T and IK. Those evidences show that the excitability of capsaicin sensitive small and medium TG neurons can be regulated by Ca(2+)-CaM-CaMKII pathway through modulating VGSCs and VGPCs. Considering the specific distribution of CaMKII and its susceptibility to many analgesic stimuli, Ca(2+)-CaM-CaMKII pathway may play an important role in the peripheral sensory transduction, especially in nociception.

Excitation of primary afferent neurons by near-infrared light in vitro

Near-infrared light therapy is an emerging neurostimulation technology, but its cellular mechanism of action remains unresolved. Using standard intracellular recording techniques, we observed that 5-10 ms pulses of 1889 nm light depolarized the membrane potential for hundreds of milliseconds in more than 85% of dorsal root ganglion and nodose ganglion neurons tested. The laser-evoked depolarizations (LEDs) exhibited complex, multiphasic kinetics comprising fast and slow components. There was no discernable difference in the LEDs in intact ganglion neurons and in acutely isolated neurons. Thus, the LED sensor seems to reside within the neuronal membrane. The near-uniform distribution of responsive neurons increased membrane conductance, and the negative reversal potential value (-41+/-2.9 mV) suggests that LED is unrelated to the activation of heat-sensitive transient receptor potential cation channel subfamily V member 1 channels. The long duration of LEDs favors an involvement of second messengers.

Glial cell line-derived neurotrophic factor acutely modulates the excitability of rat small-diameter trigeminal ganglion neurons innervating facial skin

Glial cell line-derived neurotrophic factor (GDNF) plays an important role in adult sensory neuron function. However, the acute effects of GDNF on primary sensory neuron excitability remain to be elucidated. The aim of the present study was to investigate whether GDNF acutely modulates the excitability of adult rat trigeminal ganglion (TRG) neurons that innervate the facial skin by using perforated-patch clamping, retrograde-labeling and immunohistochemistry techniques. Fluorogold (FG) retrograde labeling was used to identify the TRG neurons innervating the facial skin. The FG-labeled small- and medium-diameter GDNF immunoreactive TRG neurons, and most of these neurons also expressed the GDNF family receptor α-1 (GFRα-1). In whole-cell voltage-clamp mode, GDNF application significantly inhibited voltage-gated K + transient (I A) and sustained (I K) currents in most dissociated FG-labeled small-diameter TRG neurons. This effect was concentration-dependent and was abolished by co-application of the protein tyrosine kinase inhibitor, K252b. Under current-clamp conditions, the repetitive firing during a depolarizing pulse were significantly increased by GDNF application. GDNF application also increased the duration of the repolarization phase and decreased the duration of the depolarization phase of the action potential, and these characteristic effects were also abolished by co-application of K252b. These results suggest that acute application of GDNF enhances the neuronal excitability of adult rat small-diameter TRG neurons innervating the facial skin, via activation of GDNF-induced intracellular signaling pathway. We therefore conclude that a local release of GDNF from TRG neuronal soma and/or nerve terminals may regulate normal sensory function, including nociception.

Experience with oral mexiletine in primary erythromelalgia in children

Primary erythromelalgia is characterized by burning pain, redness, and warmth in the extremities. We present two cases of primary erythromelalgia both of whom presented with a history of several months of severe burning pain in both hands and feet. Both patients had received multiple pain medications with no improvement in symptoms. Pain was relieved by putting affected parts in ice cold water, which resulted in immersion injury of the affected parts. Both patients stopped taking part in school and social activities. We tried oral mexiletine, a class Ib antiarrythmic agent, in view of its reported role in various chronic painful conditions. Dramatic improvement was observed with its use. Both patients improved after several weeks of use, and there were fewer soaking episodes. We observed no adverse effects with mexilitine therapy.

Hyperpolarization-activated current (Ih) contributes to excitability of primary sensory neurons in rats

In various excitable tissues, the hyperpolarization-activated, cyclic nucleotide-gated current (Ih) contributes to burst firing by depolarizing the membrane after a period of hyperpolarization. Alternatively, conductance through open channels Ih channels of the resting membrane may impede excitability. Since primary sensory neurons of the dorsal root ganglion show both loss of Ih and elevated excitability after peripheral axonal injury, we examined the contribution of Ih to excitability of these neurons. We used a sharp electrode intracellular technique to record from neurons in nondissociated ganglia to avoid potential artefacts due to tissue dissociation and cytosolic dialysis. Neurons were categorized by conduction velocity. Ih induced by hyperpolarizing voltage steps was completely blocked by ZD7288 (approximately 10 µM), which concurrently eliminated the depolarizing sag of transmembrane potential during hyperpolarizing current injection. Ih was most prominent in rapidly conducting Aα/β neurons, in which ZD7288 produced resting membrane hyperpolarization, slowed conduction velocity, prolonged action potential (AP) duration, and elevated input resistance. The rheobase current necessary to trigger an AP was elevated and repetitive firing was inhibited by ZD7288, indicating an excitatory influence of Ih. Less Ih was evident in more slowly conducting Aδ neurons, resulting in diminished effects of ZD7288 on AP parameters. Repetitive firing in these neurons was also inhibited by ZD7288, and the peak frequency of AP transmission during tetanic bursts was diminished by ZD7288. Slowly conducting C-type neurons showed minimal Ih, and no effect of ZD7288 on excitability was seen. After spinal nerve ligation, axotomized neurons had less Ih compared to control neurons and showed minimal effects of ZD7288 application. We conclude that Ih supports sensory neuron excitability, and loss of Ih is not a factor contributing to increased neuronal excitability after peripheral axonal injury.

Monocyte chemoattractant protein‐1 functions as a neuromodulator in dorsal root ganglia neurons

It has previously been observed that expression of chemokine monocyte chemoattractant protein-1 (MCP-1/CC chemokine ligand 2 (CCL2)) and its receptor CC chemokine receptor 2 (CCR2) is up-regulated by dorsal root ganglion (DRG) neurons in association with rodent models of neuropathic pain. MCP-1 increases the excitability of nociceptive neurons after a peripheral nerve injury, while disruption of MCP-1/CCR2 signaling blocks the development of neuropathic pain, suggesting MCP-1 signaling is responsible for heightened pain sensitivity. To define the mechanisms of MCP-1 signaling in DRG, we studied intracellular processing, release, and receptor-mediated signaling of MCP-1 in DRG neurons. We found that in a focal demyelination model of neuropathic pain both MCP-1 and CCR2 were up-regulated by the same neurons including transient receptor potential vanilloid receptor subtype 1 (TRPV1) expressing nociceptors. MCP-1 expressed by DRG neurons was packaged into large dense-core vesicles whose release could be induced from the soma by depolarization in a Ca2+-dependent manner. Activation of CCR2 by MCP-1 could sensitize nociceptors via transactivation of transient receptor potential channels. Our results suggest that MCP-1 and CCR2, up-regulated by sensory neurons following peripheral nerve injury, might participate in neural signal processing which contributes to sustained excitability of primary afferent neurons.

A New “Kine” of Pain: MCP-1 and Sensory Neuron Excitability. Focus on “MCP-1 Enhances Excitability of Nociceptive Neurons in Chronically Compressed Dorsal Root Ganglia”

In 1998, [Hu and Xing (1998)][1] developed a method for nerve injury by chronically compressed dorsal root ganglia (CCD). Robert LaMotte and his colleagues at Yale have utilized this technique to explore the consequences of injury on nociception and excitability of primary sensory neurons. This

Upregulation of the Voltage-Gated Sodium Channel β2 Subunit in Neuropathic Pain Models: Characterization of Expression in Injured and Non-Injured Primary Sensory Neurons

The development of abnormal primary sensory neuron excitability and neuropathic pain symptoms after peripheral nerve injury is associated with altered expression of voltage-gated sodium channels (VGSCs) and a modification of sodium currents. To investigate whether the beta2 subunit of VGSCs participates in the generation of neuropathic pain, we used the spared nerve injury (SNI) model in rats to examine beta2 subunit expression in selectively injured (tibial and common peroneal nerves) and uninjured (sural nerve) afferents. Three days after SNI, immunohistochemistry and Western blot analysis reveal an increase in the beta2 subunit in both the cell body and peripheral axons of injured neurons. The increase persists for >4 weeks, although beta2 subunit mRNA measured by real-time reverse transcription-PCR and in situ hybridization remains unchanged. Although injured neurons show the most marked upregulation,beta2 subunit expression is also increased in neighboring non-injured neurons and a similar pattern of changes appears in the spinal nerve ligation model of neuropathic pain. That increased beta2 subunit expression in sensory neurons after nerve injury is functionally significant, as demonstrated by our finding that the development of mechanical allodynia-like behavior in the SNI model is attenuated in beta2 subunit null mutant mice. Through its role in regulating the density of mature VGSC complexes in the plasma membrane and modulating channel gating, the beta2 subunit may play a key role in the development of ectopic activity in injured and non-injured sensory afferents and, thereby, neuropathic pain.

Contribution of the tetrodotoxin-resistant voltage-gated sodium channel NaV1.9 to sensory transmission and nociceptive behavior.

The transmission of pain signals after injury or inflammation depends in part on increased excitability of primary sensory neurons. Nociceptive neurons express multiple subtypes of voltage-gated sodium channels (NaV1s), each of which possesses unique features that may influence primary afferent excitability. Here, we examined the contribution of NaV1.9 to nociceptive signaling by studying the electrophysiological and behavioral phenotypes of mice with a disruption of the SCN11A gene, which encodes NaV1.9. Our results confirm that NaV1.9 underlies the persistent tetrodotoxin-resistant current in small-diameter dorsal root ganglion neurons but suggest that this current contributes little to mechanical thermal responsiveness in the absence of injury or to mechanical hypersensitivity after nerve injury or inflammation. However, the expression of NaV1.9 contributes to the persistent thermal hypersensitivity and spontaneous pain behavior after peripheral inflammation. These results suggest that inflammatory mediators modify the function of NaV1.9 to maintain inflammation-induced hyperalgesia.

Beta adrenergic inhibition of capsaicin-induced, NK 1 receptor-mediated nerve growth factor biosynthesis in rat skin

Excitation of primary afferent neurons stimulates the expression of cytokines and nerve growth factor (NGF) in innervated tissues. Since NGF is a neurotrophic and immunomodulatory factor contributing to inflammatory hyperalgesia and tissue response to injury, this study was conducted in order to investigate the mechanisms by which afferent neuron stimulation by topical application of capsaicin increases NGF in the rat skin. Thereby it was sought to identify possible targets for pharmacological modulation of NGF biosynthesis. Topical capsaicin (>1 mg/ml ethanol) caused a concentration- and time-dependent increase in the concentration of NGF in rat skin. The capsaicin-induced increase of NGF was not significantly affected by indomethacin administered at a dose (2 mg/kg) that abolishes prostaglandin E 2 biosynthesis. The NGF increase was suppressed by treatment of rats with the selective tachykinin NK 1 receptor antagonist SR140333 (0.1 mg/kg), and by the beta adrenergic agonist terbutaline (0.3 mg/kg). The effect of terbutaline was reversed by the beta adrenergic antagonist propranolol (1 mg/kg). Terbutaline also inhibited the increase in NGF caused by intraplantar injection of the NK 1 receptor agonist substance P (SP), but did not significantly affect that caused by carrageenan. The results show that topical administration of capsaicin causes a primarily NK 1 receptor-dependent increase in the NGF content of rat skin, which is susceptible to inhibition by beta adrenergic agonists. These observations not only suggest regulation of skin NGF biosynthesis by afferent neuronal and adrenergic mechanisms, but also indicate possible targets for pharmacological modulation of skin NGF biosynthesis.

Onset and recovery of hyperalgesia and hyperexcitability of sensory neurons following intervertebral foramen volume reduction and restoration

Objective To investigate the relationships between L4 and L5 intervertebral foramen (IVF) stenosis (IVFS), as well as the restoration and onset and recovery of behavioral hyperalgesia and alterations in primary sensory neuron excitability. Methods IVFS was produced by surgically implanting stainless steel rods unilaterally into the intervertebral foramen at L4 and L5. The insertion of a stainless steel rod in the IVF caused IVF volume reduction, which mimics IVFS. The rods were kept for up to 14 weeks in 16 rats and 2 to 4 weeks in another 32 rats. Rod withdrawal was expected to restore the IVF volume. The rods were withdrawn in 20 rats on the 7th day and in another 20 rats on the 14th day, postoperatively. Two additional groups of control rats received no surgery or sham operation. Behavioral hyperalgesia was evidenced by the significantly decreased threshold and shortened latency of foot withdrawal to mechanical and thermal stimulation of the plantar surface. Electrophysiological intracellular recordings were obtained in vitro from L4 and/or L5 dorsal root ganglia (DRG). Results The IVFS rats exhibited a rapid-onset (≤1 day), long-lasting (10-11 weeks), mechanical, and thermal hyperalgesia. DRG neurons in each category, large-sized, medium-sized, and small-sized, from IVFS rats were more excitable than those from control rats, evaluated by the significantly decreased threshold current and action potential threshold and increased number of discharges evoked by depolarizing current and incidence of spontaneous activity. IVF volume restoration significantly reduced behavioral hyperalgesia and the increased excitability of DRG neurons. In contrast, sham surgery produced no behavioral or electrophysiological changes in the ganglion neurons. Conclusion The present study demonstrates that hyperalgesia and hyperexcitability of the primary sensory neurons can be induced following the IVF volume reduction produced by insertion of a stainless steel rod and mostly relieved by the rod withdrawal. The recovery of excitability of DRG cells to normal levels is associated with the abatement of hyperalgesia. These results support the hypothesis that increased excitability of DRG neurons is associated with the generation and maintenance of hyperalgesia and suggest that relief of the IVF stenosis, which could compress all of the normal constituents within the IVF (ie, DRG, nerve root, blood and lymph vessels, adipose, etc.), may help to alleviate chronic pain in humans.

Ion channels in airway afferent neurons

Action potentials initiated at the peripheral terminal of an afferent nerve are conducted to the central nervous system therein causing release of neurotransmitters that excite secondary neurons in the brain stem or spinal cord. Various chemicals, extremes in osmolarity and pH as well as mechanical stimuli are sensed by primary afferent nerves that innervate the airways. The processes leading to excitation of afferent nerve endings, conduction of action potentials along axons, transmitter secretion, and neuronal excitability are regulated by ions flowing through channels in the nerve membrane. Voltage-gated ion channels selective for K + and Na + ions allow the generation and conduction of action potentials and along with families of ion channels selective for other ions such as Ca 2+ or Cl − are thought to play distinctive roles in regulating neuronal excitability and transmitter secretion. Here we discuss, in general terms, the roles played by various classes of ion channels in the activation, neurotransmitter secretion and excitability of primary afferent neurons.

A Novel Persistent Tetrodotoxin-Resistant Sodium Current In SNS-Null And Wild-Type Small Primary Sensory Neurons

TTX-resistant (TTX-R) sodium currents are preferentially expressed in small C-type dorsal root ganglion (DRG) neurons, which include nociceptive neurons. Two mRNAs that are predicted to encode TTX-R sodium channels, SNS and NaN, are preferentially expressed in C-type DRG cells. To determine whether there are multiple TTX-R currents in these cells, we used patch-clamp recordings to study sodium currents in SNS-null mice and found a novel persistent voltage-dependent sodium current in small DRG neurons of both SNS-null and wild-type mice. Like SNS currents, this current is highly resistant to TTX (Ki = 39+/-9 microM). In contrast to SNS currents, the threshold for activation of this current is near 70 mV, the midpoint of steady-state inactivation is -44 +/- 1 mV, and the time constant for inactivation is 43+/-4 msec at 20 mV. The presence of this current in SNS-null and wild-type mice demonstrates that a distinct sodium channel isoform, which we suggest to be NaN, underlies this persistent TTX-R current. Importantly, the hyperpolarized voltage-dependence of this current, the substantial overlap of its activation and steady-state inactivation curves and its persistent nature suggest that this current is active near resting potential, where it may play an important role in regulating excitability of primary sensory neurons.

Sodium channels, excitability of primary sensory neurons, and the molecular basis of pain

Following nerve injury, primary sensory neurons (dorsal root ganglion [DRG] neurons, trigeminal neurons) exhibit a variety of electrophysiological abnormalities, including increased baseline sensitivity and/or hyperexcitability, which can lead to abnormal burst activity that underlies pain, but the molecular basis for these changes has not been fully understood. Over the past several years, it has become clear that nearly a dozen distinct sodium channels are encoded by different genes and that at least six of these (including at least three distinct DRG- and trigeminal neuron–specific sodium channels) are expressed in primary sensory neurons. The deployment of different types of sodium channels in different types of DRG neurons endows them with different physiological properties. Dramatic changes in sodium channel expression, including downregulation of the SNS/PN3 and NaN sodium channel genes and upregulation of previously silent type III sodium channel gene, occur in DRG neurons following axonal transection. These changes in sodium channel gene expression are accompanied by a reduction in tetrodotoxin (TTX)-resistant sodium currents and by the emergence of a TTX-sensitive sodium current which recovers from inactivation (reprimes) four times more rapidly than the channels in normal DRG neurons. These changes in sodium channel expression poise DRG neurons to fire spontaneously or at inappropriately high frequencies. Changes in sodium channel gene expression also occur in experimental models of inflammatory pain. These observations indicate that abnormal sodium channel expression can contribute to the molecular pathophysiology of pain. They further suggest that selective blockade of particular subtypes of sodium channels may provide new, pharmacological approaches to treatment of disease involving hyperexcitability of primary sensory neurons. © 1999 John Wiley & Sons, Inc. Muscle Nerve 22:1177–1187, 1999

Sodium channels, excitability of primary sensory neurons, and the molecular basis of pain.

Following nerve injury, primary sensory neurons (dorsal root ganglion [DRG] neurons, trigeminal neurons) exhibit a variety of electrophysiological abnormalities, including increased baseline sensitivity and/or hyperexcitability, which can lead to abnormal burst activity that underlies pain, but the molecular basis for these changes has not been fully understood. Over the past several years, it has become clear that nearly a dozen distinct sodium channels are encoded by different genes and that at least six of these (including at least three distinct DRG- and trigeminal neuron-specific sodium channels) are expressed in primary sensory neurons. The deployment of different types of sodium channels in different types of DRG neurons endows them with different physiological properties. Dramatic changes in sodium channel expression, including downregulation of the SNS/PN3 and NaN sodium channel genes and upregulation of previously silent type III sodium channel gene, occur in DRG neurons following axonal transection. These changes in sodium channel gene expression are accompanied by a reduction in tetrodotoxin (TTX)-resistant sodium currents and by the emergence of a TTX-sensitive sodium current which recovers from inactivation (reprimes) four times more rapidly than the channels in normal DRG neurons. These changes in sodium channel expression poise DRG neurons to fire spontaneously or at inappropriately high frequencies. Changes in sodium channel gene expression also occur in experimental models of inflammatory pain. These observations indicate that abnormal sodium channel expression can contribute to the molecular pathophysiology of pain. They further suggest that selective blockade of particular subtypes of sodium channels may provide new, pharmacological approaches to treatment of disease involving hyperexcitability of primary sensory neurons.

Alteration of Na + currents in dorsal root ganglion neurons from rats with a painful neuropathy

Increased excitability of primary sensory neurons may be important for the generation of neuropathic pain from nerve injury. The currents underlying the action potentials of these neurons are largely carried by Na +, and changes in Na + currents have been postulated to contribute to this increased excitability. Using patch clamp in whole-cell mode, we recorded Na + currents from DRG neurons freshly isolated from rats with a chronic constriction injury (CCI), an animal model of neuropathic pain. We found significant changes in Na + currents after CCI when cell size and Na + channel properties were used to segregate DRG neurons. Most changes were concentrated in small neurons (≤25 μm diameter) and in the slow, TTX-resistant current that is predominant in these cells. CCI produced two principle changes in these cells: it shifted the voltage-dependence of activation of the TTX-resistant current to more negative potentials, and it reduced the average density of this current. The decrease in density appears to be primarily due to the decrease in the number of small neurons expressing this current. The net result is a change in both activation and steady-state inactivation properties of the total Na + current to more negative potentials without a significant change in the density of the total Na + current. The change in activation properties of the TTX-resistant Na + current are similar to those produced by some hyperalgesic autacoids, and may contribute to the increase in primary afferent excitability and hyperalgesia that occurs after this lesion.

Antagonism by picrotoxin of 5-hydroxytryptamine-induced excitation of primary afferent neurons

Pharmacological studies have revealed that the effects of certain putative neurotransmitters on peripheral neurons in autonomic and sensory ganglia are very similar to the effects of these agents on primary afferent terminals in the central nervous system1,2, 4 15,17,18,23,34. Thus it was suggested that peripheral ganglia might be useful model systems for examining the 'presynaptic' actions of drugs 4,1°-1~A7. In this regard it was shown that gamma-aminobutyric acid (GABA) a transmitter which hyperpolarizes central nerve cells 6 depolarizes peripheral ganglion cells and primary afferent terminalsl,2,4-15,1s, 34. The depolarizing responses were blocked by picrotoxin and bicuculline, agents which also block neurally-evoked primary afferent depolarization and presynaptic inhibition in the spinal cord5-7A6,e4, 31. These observations provided support for the view that GABA was a presynaptic inhibitory transmitter. The present investigation was undertaken to study the response of sensory ganglion cells to 5-hydroxytryptamine (5-HT), a substance that has also been implicated in presynaptic inhibition in the spinal c0rd3,27,29,~2,~, 36. It is well known that 5-HT depolarizes and excites afferent nerves as well as peripheral ganglion cells14,2°,e5,26,28,a°, 35. In autonomic ganglia its excitatory effects are blocked by picrotoxin14, 30. The present experiments demonstrate that picrotoxin also blocks the excitatory effects of 5-HT on afferent neurons in the nodose ganglion and afferent axons in the carotid sinus nerve. Experiments were performed on 14 cats anesthetized with a mixture of diallylbarbiturate (70 mg/kg), urethane (280 mg/kg) and monoethylurea (280 mg/kg). After intubation of the trachea, the nodose ganglion, the carotid sinus nerve and vagus nerve on the left side were dissected free from underlying connective tissue. Potentials were recorded from the surface of the ganglion with silver-silver chloride electrodes connected to a resistance-coupled preamplifier. One electrode was placed in direct

Anticoagulants in the treatment of small cell carcinoma of the bronchus.

The object of this study was to see if the addition of anticoagulants to a regimen of cytotoxic drugs would improve the prognosis in patients with small cell carcinoma of the bronchus. Twenty-four patients were randomly allocated to receive chemotherapy or chemotherapy plus anticoagulants. The median survival in the group receiving the anticoagulants was not improved.