Role of the vestibular nuclear complex in facilitating the jaw-opening reflex following stimulation of the red nucleus.

According to our previous studies, stimulation of the red nucleus (RN) facilitates the low-threshold afferent-evoked jaw-opening reflex (L-JOR). It has been reported that the RN projects to the superior (SVN), lateral (LVN) and inferior vestibular (IVN) nuclei. The SVN and the LVN have reciprocal intrinsic connections with the medial vestibular nucleus (MVN). Our previous study demonstrated that stimulation of the vestibular nuclear complex (VN) modulates the L-JOR. These facts suggest that RN-induced facilitation of the L-JOR is mediated via the VN. In the present work we investigated whether electrically induced lesions of the VN, or microinjection of muscimol into the VN, affects RN-induced facilitation of the L-JOR. The L-JOR was evoked by electrical stimulation of the inferior alveolar nerve. The stimulus intensity was 1.2 times the evocation threshold. Lesions of the MVN or the LVN or the SVN, and the muscimol injection into the MVN or the LVN or the SVN, reduced the RN-induced facilitation of the L-JOR. Conversely, lesions of the IVN, and the muscimol injection into the IVN, increased the RN-induced facilitation of the L-JOR. These results suggest that the RN-induced facilitation of the L-JOR is mediated by a relay in the VN.

Suppression of the swallowing reflex by stimulation of the red nucleus.

We study whether the red nucleus is involved in control of swallowing. The swallowing reflex was induced in anesthetized rats by repetitive electrical stimulation of the superior laryngeal nerve. The electromyographic activities of the mylohyoid and thyrohyoid muscles were recorded in order to identify the swallowing reflex. Repetitive electrical stimulation applied to the red nucleus reduced the number of swallows. The onset latency of the first swallow was increased during repetitive electrical stimulation applied to the magnocellular part of the red nucleus. Microinjection of monosodium glutamate into the red nucleus also reduced the number of swallows. The onset latency of the first swallow was increased after microinjection of monosodium glutamate into the magnocellular part of the red nucleus. These results imply that the red nucleus is involved in the control of swallowing.

Role of the red nucleus in suppressing the jaw-opening reflex following stimulation of the raphe magnus nucleus.

In a previous study, we found that electrical and chemical stimulation of the red nucleus (RN) suppressed the high-threshold afferent-evoked jaw-opening reflex (JOR). It has been reported that the RN receives bilaterally projection fibers from the raphe magnus nucleus (RMg), and that stimulation of the RMg inhibits the tooth pulp-evoked nociceptive JOR. These facts imply that RMg-induced inhibition of the JOR could be mediated via the RN. The present study first examines whether stimulation of the RMg suppresses the high-threshold afferent-evoked JOR. The JOR was evoked by electrical stimulation of the inferior alveolar nerve (IAN), and was recorded as the electromyographic response of the anterior belly of the digastric muscle. The stimulus intensity was 4.0 (high-threshold) times the threshold. Conditioning electrical stimulation of the RMg significantly suppressed the JOR. A further study then examined whether electrically induced lesions of the RN or microinjection of muscimol into the RN affects RMg-induced suppression of the JOR. Electrically induced lesions of the bilateral RN and microinjection of muscimol into the bilateral RN both reduced the RMg-induced suppression of the JOR. These results suggest that RMg-induced suppression of the high-threshold afferent-evoked JOR is mediated by a relay in the RN.

Role of the lateral reticular nucleus in suppressing the jaw-opening reflex following stimulation of the red nucleus.

We found in a previous study that stimulation of the red nucleus (RN) facilitated the low-threshold afferent-evoked jaw-opening reflex (JOR) and suppressed the high-threshold afferent-evoked JOR. It has been reported that the RN projections to the contralateral lateral reticular nucleus (LRt), and stimulation of the LRt inhibits the nociceptive JOR. These facts suggest that RN-induced modulation of the JOR is mediated via the LRt. We investigated whether electrically induced lesions of the LRt, or microinjection of muscimol into the LRt, affects RN-induced modulation of the JOR. The JOR was evoked by electrical stimulation of the inferior alveolar nerve (IAN), and was recorded as the electromyographic response of the anterior belly of the digastric muscle. The stimulus intensity was either 1.2 (low-threshold) or 4.0 (high-threshold) times the threshold. Electrically induced lesion of the LRt and microinjection of muscimol into the LRt reduced the RN-induced suppression of the high-threshold afferent-evoked JOR, but did not affect the RN-induced facilitation of the low-threshold afferent-evoked JOR. These results suggest that the RN-induced suppression of the high-threshold afferent-evoked JOR is mediated by a relay in the contralateral LRt.

Modulation of two types of jaw-opening reflex by stimulation of the red nucleus.

The red nucleus (RN) is divided cytoarchitecturally into two parts, the parvicellular part (RPC) and the magnocellular part (RMC). The present study aims, first, to compare the effects of RN stimulation between low- and high-threshold afferents-evoked jaw opening reflexes (JORs), and secondly to compare the size of these effects in the RPC and RMC. Experiments were performed on rats anesthetized with urethane-chloralose. The JOR was evoked by electrical stimulation of the inferior alveolar nerve and was recorded as the electromyographic response of the anterior belly of the digastric muscle. The stimulus intensity was either 1.2 (low-threshold) or 4.0 (high-threshold) times that necessary to elicit the JOR. Conditioning electrical stimulation of the RN significantly facilitated the JOR evoked by the low-threshold afferents. On the other hand, conditioning electrical stimulation of the RN significantly suppressed the JOR evoked by the high-threshold afferents. Microinjection of monosodium glutamate into the RN also facilitated the JOR evoked by the low-threshold afferents, but suppressed that evoked by high-threshold afferents. Facilitation did not differ between the RMC and the RPC. Suppression by the RMC stimulation was significantly greater than that by the RPC stimulation. These results suggest that the RN has distinct functional roles in the control of the JOR.

Suppression of the nociceptive jaw-opening reflex by stimulation of the red nucleus.

We studied the effect of stimulation of the red nucleus (RN) on the jaw-opening reflex (JOR) in anesthetized rats. The JOR was evoked by electrical stimulation of the tooth pulp of a lower incisor, and was recorded as the electromyographic responses of the anterior belly of the digastric muscle, bilaterally. Conditioning electrical stimulation of the RN was found to suppress the JOR bilaterally. Microinjection of monosodium glutamate into the RN also suppressed the JOR bilaterally. The suppressive effect of the magnocellular part of the RN was significantly larger than that of the parvicellular part of the RN. These results imply that the RN is involved in control of the JOR evoked by noxious stimulus.

Neuronal activities of the vestibular nuclear complex during mechanically induced rhythmic jaw movements in rats.

We studied the neuronal activities of the vestibular nuclear complex (VN) neurons during rhythmic jaw movements in rats anesthetized with urethane. Rhythmic jaw movements were induced by mechanical stimulation of the palate mucosa. The firing rate of approximately 25% of VN neurons increased significantly, and that of 10% of VN neurons decreased significantly, during these rhythmic jaw movements. There was no correlation between the change in the firing rate and the phase of the rhythmic jaw movements (jaw-opening and jaw-closing phases). The neurons that were affected were intermingled in the VN. These results suggest that the VN neurons are involved in controlling jaw movements.

The rostral parvicellular reticular formation neurons mediate lingual nerve input to the rostral ventrolateral medulla.

In rats that had been anesthetized by urethane-chloralose, we investigated whether neurons in the rostral part of the parvicellular reticular formation (rRFp) mediate lingual nerve input to the rostral ventrolateral medulla (RVLM), which is involved in somato-visceral sensory integration and in controlling the cardiovascular system. We determined the effect of the lingual nerve stimulation on activity of the rRFp neurons that were activated antidromically by stimulation of the RVLM. Stimulation of the lingual trigeminal afferent gave rise to excitatory effects (10/26, 39%), inhibitory effects (6/26, 22%) and no effect (10/26, 39%) on the RVLM-projecting rRFp neurons. About two-thirds of RVLM-projecting rRFp neurons exhibited spontaneous activity; the remaining one-third did not. A half (13/26) of RVLM-projecting rRFp neurons exhibited a pulse-related activity, suggesting that they receive a variety of peripheral and CNS inputs involved in cardiovascular function. We conclude that the lingual trigeminal input exerts excitatory and/or inhibitory effects on a majority (61%) of the RVLM-projecting rRFp neurons, and their neuronal activity may be involved in the cardiovascular responses accompanied by the defense reaction.

Effects of neck muscle activities during rhythmic jaw movements by stimulation of the medial vestibular nucleus in rats.

This study first examines whether there is rhythmic activity of the neck muscles during cortically induced rhythmic jaw movements in rats anesthetized by urethane. Rhythmic jaw movements were induced by repetitive electrical stimulation of the orofacial motor cortex. An electromyogram in the splenius muscles (spEMG) showed rhythmic bursts during the jaw-opening phase, or during the transition from the jaw-opening phase to the jaw-closing phase. In the sternomastoid (stEMG), however, the electromyogram did not show any bursts during rhythmic jaw movements. A further study then examines whether stimulation of the medial vestibular nucleus (MVN) modulates the rhythmic activity of the neck muscles. Stimuli applied in the jaw-closing phase induced a transient burst in the stEMG, and the duration of activity in the spEMG was increased. Stimuli applied in the jaw-opening phase induced a transient burst in the stEMG and an inhibitory period in the spEMG. These results imply that the MVN is involved in the modulation of neck muscle activities during rhythmic jaw movements induced by stimulating the orofacial motor cortex.

Modulation of cortically induced rhythmic jaw movements in rats by stimulation of the vestibular nuclear complex.

We study whether stimulation of the vestibular nuclear (VN) complex can modulate rhythmic jaw movements in rats anesthetized by urethane. Rhythmic jaw movements were induced by repetitive electrical stimulation of the orofacial motor cortex. Stimulation of the medial vestibular nucleus (MVN) during the jaw-closing phase increased the amplitude of the jaw-closing movement. (This is not a movement that continues to closure.) Stimulation of the MVN during the jaw-opening phase disturbed the rhythm of jaw movements and induced a small jaw-closing movement. Stimulation of the superior VN (SVN) and the lateral VN (LVN) during the jaw-closing phase did not affect the amplitude of the jaw-closing movement. Stimulation of the SVN and the LVN during the jaw-opening phase increased the amplitude of the jaw-opening movement, however. Stimulation of the inferior VN during the jaw-closing and the jaw-opening phase, respectively decreased the amplitude of the jaw-closing and the jaw-opening movements. Stimulation applied outside the VN did not modulate the amplitude of the jaw movements. These results imply that the VN is involved in the modulation of rhythmic jaw movements induced by stimulation of the orofacial motor cortex.

Multi-source inputs converge on the superior salivatory nucleus neurons in anaesthetized rats.

Activation of parasympathetic nerves innervating salivary glands evokes not only salivation but also vascular responses. These parasympathetic nerves may have cardiac and/or respiratory-related activity as well as the cardiovascular sympathetic nerves that control vascular bed of salivary glands. Therefore, we investigated whether preganglionic superior salivatory nucleus (SSN) neurons projecting to the submandibular and intra-lingual ganglia exhibit pulse-related and/or respiratory-related activity, and whether they can be excited by electrical stimulation of the lingual nerve. 25% of SSN neurons were found to have pulse-related and tracheal pressure-related activities, implying that they receive cardiac and respiratory inputs. 44% of neurons exhibited only pulse-related activity, whereas 31% of the neurons had neither pulse-related nor tracheal pressure-related activity. Neurons with pulse and tracheal pressure-related activities, and those only with pulse-related activity, had B and C fibre range axons. 53% of SSN neurons received both cardiac and lingual nerve inputs. 16% of neurons recorded were found to receive only cardiac inputs, and 26% only lingual nerve inputs; whereas 5% received neither cardiac nor lingual nerve inputs. We conclude that the inputs from diverse sources converge on the SSN neurons, and they can cooperate to modulate SSN neuronal activity.

Modulation of the masseteric monosynaptic reflex by stimulation of the vestibular nuclear complex in rats.

The effect of stimulation of the vestibular nuclear complex (VN) on the masseteric monosynaptic reflex (MMR) was studied in anesthetized rats. The MMR was evoked by electrical stimulation of the mesencephalic trigeminal nucleus and was recorded, bilaterally, as the electromyographic responses of the masseter muscles. Conditioning electrical stimulation of the medial vestibular nucleus (MVN) facilitated the MMR bilaterally, as did microinjection of monosodium glutamate into the MVN. In contrast, conditioning electrical stimulation of the inferior vestibular nucleus (IVN) inhibited the MMR bilaterally. Microinjection of monosodium glutamate into the IVN also inhibited the MMR bilaterally. Conditioning electrical stimulation of the lateral and superior vestibular nuclei did not modulate the MMR. These results suggest that the MVN and the IVN are involved in modulation of the MMR and plays an important role in controlling jaw movements.

Modulation of the jaw-opening reflex by stimulation of the vestibular nuclear complex in rats.

The effect of stimulation of the vestibular nuclear complex (VN) on the jaw-opening reflex (JOR) was studied in anesthetized rats. The JOR was evoked by electrical stimulation of the inferior alveolar nerve, and was recorded as the electromyographic responses of the anterior belly of the digastric muscle, bilaterally. Conditioning electrical stimulation of the medial (MVN), lateral (LVN) and superior (SVN) vestibular nuclei facilitated the JOR bilaterally. Microinjection of monosodium glutamate into the SVN, LVN and MVN also facilitated the JOR bilaterally. In contrast, conditioning electrical stimulation of the inferior vestibular nucleus (IVN) inhibited the JOR ipsilaterally. Microinjection of monosodium glutamate into the IVN also inhibited the JOR ipsilaterally. These results suggest that the VN is involved in modulation of the JOR and plays an important role in controlling the jaw movements.

Non-NMDA and NMDA receptor agonists induced excitation and their differential effect in activation of superior salivatory nucleus neurons in anaesthetized rats.

We investigated the effects of the ionophoretic application of ionotropic non-NMDA receptor agonist (AMPA) and NMDA receptor agonist (NMDA) on extracellularly recorded and antidromically identified superior salivatory nucleus (SSN) neurons. A great majority (93%) of SSN neurons was induced to fire by ionophoretic application of AMPA, and they were classified into high firing rate (more than 6 spikes/s), and low firing rate (less than 3 spikes/s) neurons. No clear differences were found between high firing rate and low firing rate neurons according their fibre type and histological locations. Of the SSN neurons that excited by AMPA, 22% (4/18) and 50% (5/9) of the neurons also were induced to fire following ionophoretic application of the NMDA receptor agonist NMDA in different concentrations, 20 mM and 100 mM, respectively. In neurons that induced firing by AMPA and by NMDA, AMPA-evoked firings were induced by the lower intensities of applied current and had higher mean firing rates than NMDA-evoked firing. Neurons that were induced firing by AMPA and by NMDA had B fibre and C fibre axons as well as those that induced firing only by AMPA. Neurons that were fired only by AMPA were found in whole SSN area, whereas neurons that were induced firing by AMPA and by NMDA were mainly found in intermediate SSN area. In conclusion, activation of ionotoropic non-NMDA receptor has a greater excitatory effect on the SSN neurons than that of ionotropic of NMDA receptor. Our data support the view that non-NMDA receptor plays a major role, whereas NMDA receptor plays a minor role, in the activation of SSN neurons.

Changes in cortically induced rhythmic jaw movements after lesioning of the red nucleus in rats.

We study whether the red nucleus (RN) lesion can modify rhythmic jaw movements. Rhythmic jaw movements were induced by repetitive electrical stimulation of the two cortical masticatory areas (area A: the orofacial motor cortex; area P: the insular cortex). Lesions made by applied electric current in the RN were found to influence the rhythmic jaw movements induced by stimulation of A-area. The distance between the maximum and minimum jaw-opening positions was less after the lesions were induced. The duration of rhythmic jaw movements was shorter after lesioning. In contrast, lesions of the RN did not influence rhythmic jaw movements induced by stimulation of the P-area. Next, kainic acid (0.2 microl, lesion group) or phosphate-buffered saline (0.2 microl, control group) was injected into the left RN. Three days after injection, rhythmic jaw movements were induced by repetitive electrical stimulation of the A-area. The distance between the maximum and minimum jaw-opening positions in the lesion group was smaller than in the control group. The rhythmic jaw movements of the lesion group had shorter duration than the control group. These results suggest that the RN is involved in the modification of jaw movements induced by stimulation of the A-area.

Effects of pioglitazone on suppressor of cytokine signaling 3 expression: potential mechanisms for its effects on insulin sensitivity and adiponectin expression.

Pioglitazone is widely used for the treatment of diabetic patients with insulin resistance. The mechanism of pioglitazone to improve insulin sensitivity is not fully understood. Recent studies have shown that the induction of suppressor of cytokine signaling 3 (SOCS3) is related to the development of insulin resistance. Here, we examined whether the insulin-sensitizing effect of pioglitazone affects the SOCS induction. In db/db mice and high-fat-fed mice, expression of SOCS3 mRNA in fat tissue was increased compared with that in lean control mice, and pioglitazone suppressed SOCS3 levels. In 3T3-L1 adipocytes, mediators of insulin resistance such as tumor necrosis factor-alpha (TNF-alpha), interleukin-6, growth hormone, and insulin increased SOCS3 expression, which was partially inhibited by pioglitazone. The ability of pioglitazone to suppress SOCS3 induction by TNF-alpha was greatly augmented by peroxisome proliferator-activated receptor gamma overexpression. SOCS3 overexpression and tyrphostin AG490, a Janus kinase 2 inhibitor, or dominant-negative STAT3 expression partially inhibited adiponectin secretion and was accompanied by decreased STAT3 phosphorylation. Conversely, pioglitazone increased adiponectin secretion and STAT3 phosphorylation in fat tissue of db/db mice and in 3T3-L1 adipocytes. These results suggest that pioglitazone exerts its effect to improve whole-body insulin sensitivity in part through the suppression of SOCS3, which is associated with the increase in STAT3 phosphorylation and adiponectin production in fat tissue.

Chronic tumor necrosis factor-alpha treatment causes insulin resistance via insulin receptor substrate-1 serine phosphorylation and suppressor of cytokine signaling-3 induction in 3T3-L1 adipocytes.

Serine phosphorylation of insulin receptor substrate (IRS)-1 and the induction of suppressor of cytokine signaling 3 (SOCS3) is recently well documented as the mechanisms for the insulin resistance. However, the relationship between these two mechanisms is not fully understood. In this study, we investigated the involvement of SOCS3 and IRS-1 serine phosphorylation in TNFalpha-induced insulin resistance in 3T3-L1 adipocytes. TNFalpha transiently stimulated serine phosphorylation of IRS-1 from 10 min to 1 h, whereas insulin-stimulated IRS-1 tyrosine phosphorylation was inhibited only after TNFalpha treatment longer than 4 h. These results suggest that serine phosphorylation of IRS-1 alone is not the major mechanism for the inhibited insulin signaling by TNFalpha. TNFalpha stimulation longer than 4 h enhanced the expression of SOCS3 and signal transducer and activator of transcription-3 phosphorylation, concomitantly with the production of IL-6. Anti-IL-6 neutralizing antibody ameliorated suppressed insulin signaling by 24 h TNFalpha treatment, when it partially decreased SOCS3 induction and signal transducer and activator of transcription-3 phosphorylation. These results suggest that SOCS3 induction is involved in inhibited insulin signaling by TNFalpha. However, low-level expression of SOCS3 by IL-6 or adenovirus vector did not affect insulin-stimulated IRS-1 tyrosine phosphorylation. Interestingly, when IRS-1 serine phosphorylation was enhanced by TNFalpha or anisomycin in the presence of low-level SOCS3, IRS-1 degradation was remarkably enhanced. Taken together, both IRS-1 serine phosphorylation and SOCS3 induction are necessary, but one of the pair is not sufficient for the inhibited insulin signaling. Chronic TNFalpha may inhibit insulin signaling effectively because it causes both IRS-1 serine phosphorylation and the following SOCS3 induction in 3T3-L1 adipocytes.

Effect of orofacial motor cortex stimulation on neuronal activity in the red nucleus.

We studied modulation of the activities of the red nucleus (RN) neurons under to electrical stimulation of the orofacial motor cortex (OfM) in urethane-anesthetized rats. Of 57 neurons studied, 30 (53%) neurons modulated the firing patterns. The firing patterns of the RN neurons were classified into four types: an excitation (E) type (n=4), a long inhibition (LI) type (n=4), a short inhibition (SI) type (n=22), and a no-effect type (n=27). These modulated neurons were intermingled in the dorso-ventral part of the RN. Our results suggest that the RN neurons receive excitatory or inhibitory inputs from the OfM.

Rostral parvicellular reticular formation neurons projecting to rostral ventrolateral medulla receive cardiac inputs in anesthetized rats.

The rostral parvicellular reticular formation (rRFp) was explored electrophysiologically in urethane-chloralose anesthetized rats. Spontaneously-active neurons that exhibited a pulse-related activity were recorded and tested for their projections to the rostral ventrolateral medulla (RVLM). About one-third (10/29) of the rRFp neurons that exhibited a pulse-related activity were antidromically activated by RVLM stimulation with conduction velocities between 0.2-4.4m/s and fell within the B and C fibre range. A majority (8/10) of these neurons had a low (<10spikes/s) mean firing rate, whereas a small proportion (2/10) had a high (>15spikes/s) mean firing rate. These findings suggest a direct pathway from the rRFp to the RVLM and suggest that neurons projecting to the RVLM receive cardiac inputs and can modulate RVLM neuronal activity.

Modulation of cortically induced rhythmical jaw movements by stimulation of the red nucleus in the rat.

We study whether stimulation of the red nucleus (RN) can modulate rhythmical jaw movements in rats anesthetized by urethane. Rhythmical jaw movements were induced by repetitive electrical stimulation of the two cortical masticatory areas (area A: the orofacial motor cortex; area P: the insular cortex). Stimuli applied to the RN did influence rhythmical jaw movements induced by stimulation of the A-area. Stimuli applied in the jaw-closing phase increased the amplitude of the jaw-closing movement. Stimuli applied in the jaw-opening phase disturbed the rhythm of jaw movements and induced a small jaw-closing movement. Stimuli applied to the RN did not influence rhythmical jaw movements induced by stimulation of the P-area. These results indicate that the RN is involved in the modulation of rhythmical jaw movements induced by stimulation of the A-area.