Effects of pharyngeal water stimulation on swallowing behaviors in healthy humans.

The aims of the present study were to determine whether the interval between swallows and the electromyographic (EMG) burst patterns of the suprahyoid muscles is affected by peripheral inputs during swallowing. Eighteen normal adults were asked to perform repetitive voluntary swallowing as quickly as possible, and three variables of swallowing were measured and evaluated, i.e., the swallowing intervals and the time interval between the onset and peak (rising time) and between the peak and offset (falling time) of the suprahyoid EMG burst. During recording, pharyngeal fluid infusion was applied with distilled water or 0.3 M NaCl solution at a very slow infusion rate (0.2 mL/min). The former and latter were used to activate and inhibit the excitation of water-sensitive receptors in the pharynx, respectively. The swallowing interval was significantly shorter during infusion of water than during infusion of NaCl solution. The rising time was also significantly shorter during infusion of water than during infusion of NaCl solution. There was a linear positive correlation between these values and facilitatory effects: the longer either the swallowing interval or rising time with infusion of 0.3 M NaCl solution, the stronger the facilitation of swallowing by the activation of water receptors. Facilitatory effects on the swallowing interval and rising time showed a linear correlation. It is suggested that weak liquid stimulation changed sensory inputs into the swallowing center and synchronously modulated the swallowing interval and time interval between the onset and peak of the EMG burst.

Effect of stimulation of the laryngopharynx with water and salt solutions on voluntary swallowing in humans: characteristics of water receptors in the laryngopharyngeal mucosa.

Stimulation of water receptors in the laryngopharynx (LP) with water facilitates voluntary swallowing in humans. Based on measures of swallowing intervals (SIs) in repetitive swallowing, we investigated characteristics of laryngopharyngeal water receptors in humans. Healthy adult volunteers were instructed to perform repetitive swallowing as quickly as possible during infusion of a solution into the LP. Infusion of water shortened SI, suggesting that water excites water receptors. Infusion of 0.3 M NaCl solution prolonged SI, suggesting that the NaCl solution inhibits activity of water receptors. SI increased with increasing concentration of NaCl. Anion or cation substitutions indicated that excitation of water receptors is due to absence or reduced concentration of Cl(-). With diminution of peripheral inputs, cortical inputs would play a dominant role in voluntary swallowing. With infusion of a nonstimulating solution (0.3 M NaCl at 0.2 mL/min), SI varied greatly from subject to subject, suggesting that the ability of central regulation of swallowing to initiate repetitive voluntary swallowing varies among subjects. Facilitation of swallowing by chemosensory inputs from water receptors appeared strongly in subjects with longer SI with infusion of the nonstimulating solution. It appears that chemosensory activation compensates for the difficulty in initiating swallowing via the central neural mechanism.

Facilitation of voluntary swallowing by chemical stimulation of the posterior tongue and pharyngeal region in humans.

In this study, we investigated the functional difference between chemical stimulations of the posterior tongue (PT) and pharyngeal region (PR) for facilitation of voluntary swallowing in humans. The PT or PR stimulation consisted of infusion of water (distilled water), 0.3 M NaCl solution or olive oil (non-chemical stimulant) into the PT or the PR through a fine tube at a very slow infusion rate (0.2 ml/min). Water was used as a stimulant of water receptors. A solution of 0.3 M NaCl was used as an inhibitor of the response of water receptors and as a stimulant of salt taste receptors. Excitation of the mucosal receptors would facilitate voluntary swallowing and diminution of sensory inputs from the oral mucosa would induce difficulty in swallowing. Swallowing intervals (SIs) during voluntary swallowing were measured by submental electromyographic activity. Infusion of water into the PR shortened SI (facilitation of swallowing) and infusion of 0.3 M NaCl or olive oil into the same region prolonged it (difficulty in swallowing). On the other hand, infusion of water into the PT prolonged SI and infusion of 0.3 M NaCl into the same region shortened it. The results suggest that water receptors are localized in the PR and that salt taste receptors are almost absent in the PR and present in the PT. With diminution of sensory inputs from the oral mucosa, central inputs would play a dominant role in initiating swallowing voluntarily, and SI would be prolonged. With weak stimulation (infusion of 0.3 M NaCl into the PR or infusion of water into the PT), SI was prolonged and inter-individual variation in SI was pronounced, suggesting that the ability of the central regulation of swallowing to perform repetitive voluntary swallowing varies among subjects. With stimulation of water receptors or salt taste receptors, SI was shortened and inter-individual variation in SI was moderate, suggesting that sensory inputs are important for performing voluntary swallowing smoothly and that the sensory inputs compensate for the difficulty in performing swallowing caused by the central mechanism.

Mechanism of enhancement of the responses of the frog glossopharyngeal nerve to electrolytes by enhancers.

In frogs, the responses of the glossopharyngeal nerve (GL) to NaCl are enhanced after treatment of the tongue with 8-anilino-1-naphthalene-sulfonic acid (ANS), a hydrophobic probe for biological membranes. The enhancement by ANS treatment has been explained by removal of Ca2+ from the receptor membrane treated with ANS. To explore the mechanism of enhancement by ANS treatment, we recorded neural responses from the frog GL. After ANS treatment, treatment with 10 mM CaCl2 prior to stimulation of NaCl did not affect the enhanced responses to 100 mM NaCl. The response to a relatively high concentration of CaCl2 (50 mM) was enhanced after ANS treatment. It is difficult to interpret these neural events in terms of modulation of the responses by membrane-bound calcium. The presence of NiCl2 in stimulating solution is known as an enhancer. Neural events after ANS treatment were similar to those caused by NiCl2. Our previous studies have demonstrated that enhancement of the responses to electrolytes by NiCl2 is due to modulation of the responses of water fibers in the GL. Water fibers are characterized by sensitivity to water or CaCl2, and they also respond to relatively high concentrations of electrolytes such as NaCl and choline Cl. Using a suction electrode method, we recorded unitary impulses from single water fibers. The ANS treatment led greatly enhanced responses to NaCl or choline Cl in water fibers, suggesting that enhancement by the ANS treatment is due to modulation of the responses of water fibers as well as enhancement by NiCl2. It appears that distinct receptors for each separate cation responsible for the neural responses in water fibers interact with a membrane element that is affected by ANS or Ni2+.

Membrane excitability of wing and rod cells in frog taste discs following denervation.

The frog tongue has a disc-shaped taste organ (taste disc) on the top of fungiform papillae. The taste disc contains two types of cells, wing cells with a sheet-like apical process and rod cells with a rod-like apical process. Both wing and rod cells can produce action potentials. Unlike the taste buds of mammals, frog taste discs do not degenerate over a long period after denervation. Here we report that the shapes of wing and rod cells isolated from taste discs in the bullfrog (Rana catesbeiana) remained unchanged 1 month after cutting bilateral glossopharyngeal nerves. By applying the whole cell patch-clamp technique to isolated wing and rod cells, we found voltage-dependent inward currents and outward currents and action potentials in denervated frogs as seen in normal frogs. These results suggest that the maintenance of morphological integrity and electrical excitability of taste cells does not require a nerve supply in frogs.

Effect of electrical stimulation of the internal capsule on nociceptive neurons responding to orofacial stimuli in the medullary dorsal horn of the rat.

Internal capsule (IC) stimulation has been used clinically to alleviate central pain. However, the neuronal mechanisms underlying pain relief by IC stimulation are poorly understood. In order to elucidate the analgesic mechanism, the effect of IC conditioning stimulation on nociceptive neurons in the rat medullary dorsal horn was investigated in the present study. Rats were anaesthetized with N(2)O-O(2) (2:1) and 0.5% halothane and were immobilized with pancuronium bromide. Two kinds of nociceptive neurons, wide dynamic range (WDR) and nociceptive specific (NS) neurons, responding to noxious stimulations of the face and oral structures were recorded in the trigeminal caudal nucleus and the medial reticular subnuclei. A test stimulus with a single rectangular pulse (2ms in duration, 5-70V) was applied to the centre of the receptive field. Responses in 55.9% of the WDR neurons and in 60% of the NS neurons were inhibited by conditioning stimuli to the ipsilateral IC with trains of 33 pulses (100-300microA) at 330Hz. The percents of peak inhibitory effects on WDR neurons and NS neurons were 78.1+/-25.0% (n=19) and 89.0+/-13.6% (n=3), respectively. The inhibitory effect continued for conditioning-test intervals of up to 500ms. Effective sites for conditioning stimulation were concentrated in the lateral side of the IC. These findings suggest that modulation of nociceptive transmission by IC stimulation occurs at second-order neurons via a presynaptic phenomenon by corticofugal fibers in the IC.

Voltage-gated inward currents of morphologically identified cells of the frog taste disc.

We used the patch clamp technique to record from taste cells in vertical slices of the bullfrog (Rana catesbeiana) taste disc. Cell types were identified by staining with Lucifer yellow in a pipette after recording their electrophysiological properties. Cells could be divided into the following three groups: type Ib (wing) cells with sheet-like apical processes, type II (rod) cells with single thick rod-like apical processes and type III (rod) cells with thin rod-like apical processes. No dye-coupling was seen either between cells of the same type or between cells of different types. We focused on the voltage-gated inward currents of the three types of cells. Type Ib and type II cells exhibited tetrodotoxin (TTX)-sensitive voltage-gated Na+ currents. Surprisingly, type III cells showed TTX-resistant voltage-gated Na+ currents and exhibited a lack of TTX-sensitive Na+ currents. TTX-resistant voltage-gated Na+ currents in taste cells are reported for the first time here. The time constant for the inactivating portion of the voltage-gated inward Na+ currents of type III cells was much larger than that of type Ib and type II cells. Therefore, slow inactivation of inward Na+ currents characterizes type III cells. Amplitudes of the maximum peak inward currents of type III cells were smaller than those of type Ib and type II cells. However, the density (pA/pF) of the maximum peak inward currents of type III cells was much higher than that of type Ib cells and close to that of type II cells. No evidence of the presence of voltage-gated Ca2+ channels in frog taste cells has been presented up to now. In this study, voltage-gated Ba2+ currents were observed in type III cells but not in type Ib and type II cells when the bath solution was a standard Ba2+ solution containing 25 mM Ba2+. Voltage-gated Ba2+ currents were blocked by addition of 2 mM CoCl2 to the standard Ba2+ solution, suggesting that type III cells possess the voltage-gated Ca2+ channels and they do classical (calcium-influx) synaptic transmission. It appears that type III cells are taste receptor cells.

Responses of diencephalic nociceptive neurones to orofacial stimuli and effects of internal capsule stimulation in the rat.

The effect of conditioning stimulation of the internal capsule on nociceptive neurones in the rat diencephalon was investigated. The animals were anaesthetised with N(2)O/O(2) (2:1) and 0.5% halothane, and immobilised with pancuronium bromide. Nociceptive neurones responding to noxious stimulation of the face and oral structures were recorded in the ventral posteromedial nucleus, posterior group and zona incerta. These neurones were classified into wide dynamic range and nociceptive-specific types. Functional segregation of these nociceptive neurones was not apparent within the nucleus or between nuclei. A test stimulus with a single rectangular pulse (5-70 V) was applied to the centre of the receptive field; the nociceptive neurones exhibited short- and/or long-latency responses. Both responses in about 45% of the nociceptive neurones were inhibited by conditioning stimuli to the contralateral internal capsule with trains of 33 pulses (300 microA) at 330 Hz. The percent inhibitory effects on the nociceptive neurones of each area were 68.0+/-14.8% (n = 6) in the ventral posteromedial nucleus, 72.8+/-12.4% (n = 4) in the posterior group and 61.5+/-7.5% (n = 4) in the zona incerta. Effective sites for conditioning stimulation were concentrated in the lateral side of the internal capsule, through which the corticofugal fibres from the somatosensory cortex pass. These findings suggest that the transmission of nociceptive information to the diencephalon is modulated by stimulation of the internal capsule at the level of the trigeminal sensory complex in the brainstem. They might provide a novel way to elucidate the neurophysiological basis for antinociception by stimulation of the internal capsule observed in clinical studies.