Influences of the sympathetic and parasympathetic nerve transection on cardiovascular reflexes induced by volleys in the IXth nerve fibers of rat.

The effect of vagotomy and sympathectomy on cardio-acceleratory and arterial hypertensive reflexes evoked by electrical stimulation of the rat glossopharyngeal (IXth) nerve was studied in relation to changes in baseline heart rate and arterial blood pressure. Uni- and bi-lateral transection of the cervical vagal trunk brought about augmentation of baseline heart rate, accompanied by a depression in reflex tachycardia; the amount of depression being inversely related to that of an increase in baseline heart rate. The latter increased more after a right vagotomy than after a left vagotomy. No appreciable change in reflex hypertension as well as in baseline blood pressure was observed by different types of vagotomy. For unilateral sympathectomy, semilunar cordotomy caudal to the obex was performed. It was found that right semilunar cordotomy significantly depressed the magnitude of cardio-acceleratory and arterial hypertensive reflexes in association with a significant decrease in baseline heart rate, though there was a less pronounced decrease in baseline blood pressure. The result obtained by a left semilunar cordotomy was similar to that of a right semilunar cordotomy, except that the decrease in reflex tachycardia was very small and statistically insignificant. Thus, the efferent activities in vagus and sympathetic nerves were more effective on the right than on the left side, in modifying reflex tachycardia and baseline heart rate, whereas, right and left sympathetic efferent outflows were equally effective in depressing the reflex increase in blood pressure.

Cardiovascular responses to gustatory and mechanical stimulation of the nasopharynx in rats.

The effects of natural (mechanical and gustatory) stimulation of the nasopharynx or electrical stimulation of the pharyngeal branch of the glossopharyngeal (PH-IXth) nerve on the changes in heart rate (HR) and arterial blood pressure (BP) were investigated in paralyzed and anesthetized rats. Afferent responses in the PH-IXth nerve were also investigated. Electrical stimulation of the PH-IXth nerve elicited a tachycardia and an increase in BP. Among the gustatory (1.0 M NaCl, 0.03 M HCl, 0.03 M QHCl, 1.0 M sucrose, H2O, and 0.9% NaCl) and mechanical stimuli applied to the nasopharynx, 1.0 M sucrose and 0.9% NaCl were ineffective in changing HR and BP; the rest of the stimuli were strongly effective as was the case with electrical stimulation of the PH-IXth nerve. Responses were evoked in the PH-IXth nerve by nasopharyngeal stimulation with the stimuli which were effective in producing cardiovascular responses. On the other hand, 1.0 M sucrose and 0.9% NaCl, which were ineffective stimuli for cardiovascular responses, did not produce any response in the PH-IXth nerve. There was a high correlation between the magnitude of the responses in the PH-IXth nerve and those of the cardiovascular system. These results indicate that gustatory and mechanical information carried in the PH-IXth nerve innervating the nasopharynx plays an important role in cardiovascular regulation as well as the sense of taste.

Power spectrum analysis of heart rate fluctuations and respiratory movements associated with cooling the human skin.

A length of respiratory movement and instantaneous heart rate was divided into 32 segments to adopt computerized power spectrum analysis. An array of frequency spectrum across all the segments was then constructed to demonstrate sequential changes in the respiratory rhythm (0.4 Hz), low (less than 0.1 Hz) and high (0.2-0.4 Hz)-frequency components of heart rate fluctuations associated with lowering the skin temperature. A significant increase in the power for the low and high frequency components of heart rate fluctuations accompanying bradycardia occurred by cooling the face to 0 and 10 degrees C, while cooling the foot to 0 degrees C yielded a significant increase and decrease in the power for the former and latter component, respectively, without change in heart rate. The spectral power for respiratory movements was not altered by these stimuli. The maximal change in the power for low- and high-frequency components as well as in the heart rate during cooling the face and foot to 0 degrees C was found to correlate negatively with the relative resting magnitude of the respective parameter in the pre-stimulus period. Similar relationship existed only with the power for low-frequency component of heart rate fluctuations when lowering the temperature of foot to 10 degrees C. The power spectrum array employed in this study thus enabled us to estimate the effectiveness of skin cooling on the three hemodynamic parameters.

Receptive fields and gustatory responsiveness of frog glossopharyngeal nerve. A single fiber analysis.

Receptive fields and responsiveness of single fibers of the glossopharyngeal (IXth) nerve were investigated using electrical, gustatory (NaCl, quinine HCl, acetic acid, water, sucrose, and CaCl2), thermal, and mechanical stimulation of the single fungiform papillae distributed on the dorsal tongue surface in frogs. 172 single fibers were isolated. 58% of these fibers (99/172) were responsive to at least one of the gustatory stimuli (taste fibers), and the remaining 42% (73/172) were responsive only to touch (touch fibers). The number of papillae innervated by a single fiber (receptive field) was between 1 and 17 for taste fibers and between 1 and 10 for touch fibers. The mean receptive field of taste fibers (X = 6.6, n = 99) was significantly larger than that of touch fibers (X = 3.6, n = 73) (two-tailed t test, P less than 0.001). In experiments with natural stimulation of single fungiform papillae, it was found that every branch of a single fiber has a similar responsiveness. Taste fibers were classified into 14 types (Type N, Q, A, NA, NCa, NCaA, NCaW, NCaAW, NCaWS, NQ, NQA, NQAS, NQWarm, Multiple) on the basis of their responses to gustatory and thermal stimuli. The time course of the response in taste fibers was found to be characteristic of their types. For example, the fibers belonging to Type NQA showed phasic responses, those in Type NCa showed tonic responses, etc. These results indicate that there are several groups of fibers in the frog IXth nerve and that every branch of an individual fiber has a similar responsiveness to the parent fiber.

The ability of gustatory stimuli to modify the cardiac sympathetic and vagus nerve activities.

The effects of chemical stimulation of the oro-lingual mucosa on the heart rate and arterial blood pressure were studied together with changes in the cardiac nerve activities in the rat. Of four qualities of basic taste stimuli, only HCl and NaCl but not quinine and sucrose were effective in increasing the heart rate and blood pressure. These changes in the cardiovascular system were associated with an increase in the efferent cardiac sympathetic activity and a decrease in the vagal activity, although they were no longer appreciable after transection of the glossopharyngeal nerve.

Taste responses of Purkinje cells in the frog cerebellum.

Fifty-nine Purkinje cells that responded to electrical stimulation of the glossopharyngeal (IXth) nerve with complex and/or simple spikes were isolated in the frog cerebellum. For these 59 Purkinje cells, changes in the complex and simple spike activity during taste stimulation of the tongue (42 cells for NaCl and 17 for quinine) were investigated. Of 42 Purkinje cells, 23 (54.8%) showed excitatory changes in simple and/or complex spike discharge rate during NaCl stimulation, and the remaining 19 (45.2%) showed no response. On the contrary, only a few Purkinje cells (2 of 17 cells, 11.8%) showed an excitatory change in simple or complex spike discharge rate during quinine stimulation. These results demonstrate that gustatory information influences cerebellar Purkinje cell activity.

Response properties of cerebellar neurons to stimulation of the frog glossopharyngeal nerve and tongue.

The cerebellum receives information from many kinds of sensory organs (muscle, somatosensory, auditory, vestibular, visual) as well as from the autonomic system. The cerebellum presumably has a role in the control of tongue movement and salivary secretion. However, the relationship between cerebellar neuron activity and tongue sensation has not been investigated previously. In the present study, negative cerebellar field potentials in the molecular layer and single unit responses of Purkinje cells induced by electrical stimulation of the bullfrog glossopharyngeal (IXth) nerve or tongue surface were investigated. The interaction between IXth nerve stimulation and natural (taste and touch) stimulation of the tongue in their effects on cerebellar neuron activity were investigated. The negative field potentials were potentiated by a brief train of electrical pulses to the tongue or IXth nerve. With electrical stimulation of the tongue surface, several fungiform papillae were needed to elicit cerebellar field potentials. The latency of Purkinje cells following IXth nerve stimulation was 44.4-53.6 msec for complex spikes, whereas for simple spikes two maxima were seen, with mean values at 33.9-36 msec and 96.8 msec. A preceding electrical stimulation of the IXth nerve depressed the negative field potentials or Purkinje cell complex spike responses induced by test stimulation of the IXth nerve. These depressive effects were also seen following a preceding natural stimulation of the tongue and were dependent upon the type of preceding stimulation. The depressive effects were produced by preceding stimulation with NaCl, CaCl2, water, and touch, but not with quinine and acetic acid stimulation. These results clearly demonstrate that gustatory and tactile signals from the tongue can influence cerebellar neuron activity.

Origin of the climbing fibers activated by glossopharyngeal nerve stimulation in the frog.

The origin of climbing fibers activated by electrical stimulation of the frog's glossopharyngeal (IXth) nerve was investigated using histological and electrophysiological technique. At the molecular layer near the Purkinje cell layer, where the maximum negative cerebellar field potential could be recorded following electrical stimulation of the IXth nerve, horseradish peroxidase (HRP) was iontophoretically injected through the tip of the recording micropipette. The HRP labeled cells were seen in the contralateral inferior olive (IO). In some cases, a small number of HRP-labeled cells were seen in the ipsilateral IO. Labeled cells were not found in the other areas of the brain stem. After electrolytic lesion of the contralateral IO, the negative cerebellar field potential which would be recorded in the molecular layer following electrical stimulation of the IXth nerve had almost ceased. These results demonstrate that the climbing fibers activated by the IXth nerve stimulation have their origin in the contralateral IO.

Multimodal responses of taste neurons in the frog nucleus tractus solitarius.

The responses of 216 neurons in the nucleus tractus solitarius (NTS) of the American bullfrog were recorded following taste, temperature, and tactile stimulation. Cells were classified on the basis of their responses to 5 taste stimuli: 0.5 M NaCl, 0.0005 M quinine-HCl (QHCl), 0.01 M acetic acid, 0.5 M sucrose, and deionized water (water). Neurons showing excitatory responses to 1, 2, 3, or 4 of the 5 kinds of taste stimuli were named Type I, II, III, or IV, respectively. Cells whose spontaneous rate was inhibited by taste and/or tactile stimulation of the tongue were termed Type V. Type VI neurons were excited by tactile stimulation alone. Of the 216 cells, 115 were excited or inhibited by taste stimuli (Types I-V), with 35 being Type I, 34 Type II, 40 Type III, 2 Type IV and 4 Type V. The remaining 101 cells were responsive only to tactile stimulation (Type VI). Of those 111 cells excited by taste stimulation (Types I-IV), 106 (95%) responded to NaCl, 66 (59%) to acetic acid, 44 (40%) to QHCl, 10 (9%) to water, and 9 (8%) to warming. No cells responded to sucrose. Of the 111 cells of Types I-IV, 76 (68%) were also sensitive to mechanical stimulation of the tongue. There was some differential distribution of these neuron types within the NTS, with more narrowly tuned cells (Type I) being located more dorsally in the nucleus than the more broadly tuned (Type III) neurons. Cells responding exclusively to touch (Type VI) were also more dorsally situated than those responding to two or more taste stimuli (Types II and III).

Selective depressant action of antidromic impulses on gustatory nerve signals.

The depressant action of antidromic volleys of impulses on gustatory nerve signals from the tongues of bullfrogs was studied. Electrical stimulation of the glossopharyngeal nerve at a rate of 100 Hz for 10 s and at supramaximal intensity slightly depressed the integrated glossopharyngeal nerve responses to quinine and to mechanical taps to the tongue. The same antidromic stimuli resulted in a 30-40% reduction in the responses to salt, acid, water, and warmed saline, but depressed greater than 80% of the afferent impulses firing spontaneously. The magnitude of responses to quinine and NaCl and the number of spontaneous discharges decreased gradually with an increase in either the frequency or the duration of antidromic stimuli. Similar results were obtained with intensities above the threshold for exciting gustatory and slowly adapting mechanosensitive fibers. The time required to recover from termination of the antidromic stimuli to two-thirds of the maximal amount of depression ranged between 6 and 7 min, with no significant differences among the depressions. The possible mechanisms involved in the antidromic depression of gustatory nerve signals are discussed.

Responses of cerebellar cortex to electrical stimulation of the glossopharyngeal nerve in the frog.

In the frog cerebellar cortex, electrical stimulation of the glossopharyngeal (IXth) nerve induced negative field potentials with a peak latency of 57 ms whose distribution was bilateral with ipsilateral predominance. The site where the maximum negativity was induced by IXth nerve stimulation was histologically located within the molecular layer near the Purkinje cell layer. In extracellular recording, electrical stimulation of the IXth nerve induced complex and/or simple spike discharges of Purkinje cells. Such evoked potentials and unitary spikes in the cerebellum were attributed to the excitation of the IXth nerve afferents of higher threshold which are mainly composed of fibers sensitive to taste stimulation. These results suggest that gustatory information projects to the cerebellum, as well as those of other kinds of senses, such as touch, visual and auditory sensation.

Gustatory signal processing in the glossopharyngeo-hypoglossal reflex arc of the frog.

The relationship between the gustatory input and motor output in the glossopharyngeo-hypoglossal reflex was analyzed on the basis of neuronal activities in the solitary tract and hypoglossal motor nuclei of bullfrogs. Concentration-response relations for NaCl, quinine and acetic acid, obtained from the glossopharyngeal (IXth) nerve and simultaneously recorded from the hypoglossal (XIIth) nerve, were expressed relative to the response of each nerve to 1 M NaCl. Compared with a relatively small amount of the afferent input for acid, the reflex motor output was much larger in the relative value. A similarly high output relation was obtained for warmed saline but not for quinine and cooled saline. Although the responsiveness of the nucleus tractus solitarius neurons to 1 M NaCl and 1 mM quinine was not significantly different from that of the hypoglossal motoneurons, responses to 10 mM acetic acid were greater in the latter neurons than in the former by a factor of about 5.2. These phenomena were consistent with those in the peripheral nerves. The solitary tract neurons responsive to NaCl, quinine and acid showed both the phasic and tonic components of discharges. According to classification by a transiency index, the discharge mode became more phasic for the hypoglossal motoneurons responsive to NaCl and quinine, but more tonic for those responsive to acid. The above-mentioned chemoreflex is thus regulated by the intrinsic neural network which sends signals to the XIIth nerve after modifying not only the amount but also the temporal pattern of gustatory nerve signals for a particular taste.

Effect of antidromic stimulation of the glossopharyngeal nerve on afferent discharges occurring with and without sensory stimulation of the frog tongue.

Repetitive stimulation of the bullfrog glossopharyngeal nerve at a rate of 100 Hz for 10 s and at a supra-maximal intensity resulted in little depression of the glossopharyngeal nerve responses to application of quinine and mechanical taps to the tongue, in a 30-40% decrease in the responses to salt, acid, water and warmed saline and in a 80% decrease in the number of spontaneous discharges from the tongue. Such a selective depression existed throughout changes in the frequency and the duration of the antidromic stimuli. The phenomena were attributed to antidromic impulses in high-threshold glossopharyngeal nerve afferents.

Intraganglionic distribution of the primary afferent neurons in the frog glossopharyngeal nerve and its transganglionic projection to the rhombencephalon studied by HPR method.

Anterograde transport of horseradish peroxidase (HRP) along the bullfrog IXth nerve was studied 6-16 days after application of HRP to the cut end of either the IXth nerve trunk or its distal 2 branches. The jugular and IXth nerve ganglia attached to the rhombencephalon were removed after fixation and serial sections of 50 microns in thickness were stained by the Graham and Karnovsky method. Of all the primary afferent neurons in the IXth nerve, 62% of the cell bodies were distributed within the IXth nerve ganglion, the remaining 38%, within the jugular ganglion. Similar distribution was found with the cells belonging to each of the IXth nerve branches. A part of the transganglionic IXth nerve fibers entering the medulla oblongata ascended to the cerebellar peduncle while the majority descended along the fasciculus solitarius. Some of the descending fibers in the fasciculus extended to the dorsal field of the spinal cord at the third spinal nerve, while some others run to join the descending tract of trigeminal nerve.

Surface and intramedullary potentials evoked by stimulation of the glossopharyngeal nerve in frogs.

The bulbar potentials evoked by afferent volleys in the bullfrog glossopharyngeal nerve and in its 2 distal branches were studied. Following supramaximal electric stimulation of the peripheral nerve, the potential consisting of 2 triphasic deflections (S1 and S2) of presynaptic origin and 4 postsynaptic negative waves, N1, N2, N3 and N4, having the peak latency of 5, 8, 30 and 80 ms, respectively, was obtained along the nucleus fasciculus solitarius. By lowering the stimulus intensity to the threshold for exciting mechanosensitive fibers, only S1 followed by N1, N3 and N4 was produced, whereas, at higher intensities, S2 which accompanied by N2 became apparent. N1 and N2 waves were distributed over the bulbar dorsal surface with the maximal amplitude at 1-2 mm rostral to the obex and 0.5-1 mm lateral from the midline, the negativity being found maximal at the depth 0.5-1 mm from the surface. The surface-recorded N2 potential evoked by stimulation of the medial branch distributed caudal to that produced by stimulation of the lateral branch. Of intramedullary-recorded 4 negative waves, only N1 caused by volleys in the lateral branch distributed deeper layer than the one evoked by those in the other branch.

Spatial distribution of somatosensory responses evoked by tapping the tongue and finger in man.

Early components of the somatosensory potential (SEP) evoked by tactile stimulation of the tongue were recorded from the scalp in 7 normal subjects and compared with those resulting from taps on the middle finger. (1) Unipolar but not bipolar recording of SEP to tapping the tongue failed to differentiate the early components from large EEG deflections of myogenic origin. No such contamination was seen in either unipolar or bipolar recording of SEP following taps on the finger. (2) The early components evoked by tapping the tongue consisted of 4 main deflections; the first negative wave (N1) peaking at about 13 msec after the stimulus was followed by the first positive (P1), second negative (N2) and second positive (P2) wave peaking at about 23, 32 and 44 msec, respectively. Each wave had a shorter latency by several msec than the corresponding wave elicited by taps on the finger. (3) To demonstrate the spatial distribution of a SEP wave over the scalp, a method of amplitude transformation of bipolar records was developed. Topography of the transformed amplitudes of P1 wave, thus obtained while tapping the tongue differed from that obtained while tapping the finger; the former indicated bilateral distribution of the P1 with the maximal amplitude at contralateral (T3 or T5) and ipsilateral (T4 or T6) temporal loci, whereas, in the latter, the distribution was most prominent over the contralateral posterior quadrant with the maximum at parietal locus (C3). P1 wave in SEP thus appeared to represent excitation of the primary cortical neurons responding to tactile stimulation of the different body areas.

Frog's tongue receptive areas: neural organization and gustatory function.

Lateral and medial branch of the frog's IXth nerve innervates rostral third and caudal two-thirds of the tongue surface, respectively. The amounts of gustatory signals in these branches differ in proportion to the area they supply.

Depression of lingual tactile sensation during chemical stimulation of the human tongue.

The subjectively estimated magnitude of human tactile sensation elicited by tapping the tongue was depressed during taste stimulation by 3 and 6% NaCl solution, 0.1% quinine and 0.5% acetic acid. No significant change in the tactile sensation occurred during flow of 2-20% sucrose solution. The result may be attributed to presynaptic inhibition exerted by gustatory onto lingual tactile afferents.