Takeshi Ishide

Glutamate neurotransmission and nitric oxide interaction within the ventrolateral medulla during cardiovascular responses to muscle contraction.

We previously reported that nitric oxide, within the RVLM and CVLM, plays an opposing role in modulating cardiovascular responses during static muscle contraction [B.J. Freda, R.S. Gaitonde, R. Lillaney, A. Ally, Cardiovascular responses to muscle contraction following microdialysis of nitric oxide precursor into ventrolateral medulla, Brain Res. 828 (1999) 60-67]. In this study, we determined whether the effects of administering L-arginine, a precursor for the synthesis of nitric oxide, and N(G)-monomethyl-L-arginine (L-NMMA), a nitric oxide synthase inhibitor, into the rostral (RVLM) and caudal (CVLM) ventrolateral medulla on cardiovascular responses elicited during static muscle contraction were mediated via an alteration of localized glutamate concentrations using microdialysis techniques. In experiments within the RVLM (n=8), muscle contraction increased MAP and HR by 21+/-2 mmHg and 22+/-3 bpm, respectively. Glutamate increased from 1.1+/-0.4 to 4.4+/- 0.6 ng/5 microl measured from bilateral RVLM areas. Microdialysis of L-arginine (1.0 microM) for 30 min attenuated the contraction-evoked increases in MAP, HR, and glutamate levels. After subsequent microdialysis of L-NMMA (1.0 microM) into the RVLM, contraction augmented the pressor and tachycardic responses and glutamate release. In experiments within CVLM (n=8), muscle contraction increased MAP and HR by 22+/-3 mmHg and 20+/-2 bpm, respectively. Glutamate increased from 0.8+/-0. 4 to 3.6+/-0.6 ng/5 microl measured from the CVLM. L-Arginine augmented the cardiovascular responses and glutamate release and L-NMMA attenuated all the effects. Results suggest that nitric oxide within the RVLM and CVLM plays opposing roles in modulating cardiovascular responses during static exercise via decreasing and increasing, respectively, extracellular glutamate levels.

Rostral ventrolateral medulla opioid receptor activation modulates glutamate release and attenuates the exercise pressor reflex

We previously reported that the administration of [D-Ala(2)]methionine enkephalinamide (DAME), an opioid receptor agonist, into the rostral (RVLM) but not into the caudal ventrolateral medulla (CVLM), attenuated increases in mean arterial pressure (MAP) and heart rate (HR) during static muscle contraction that had been blocked by prior microdialysis of the opioid receptor antagonist, naloxone [Am. J. Physiol. 274 (1998) H139-H146]. In this study, we determine whether this RVLM-mediated opioidergic-modulation of cardiovascular responses is associated with localized changes in extracellular concentrations of glutamate, an excitatory amino acid, using microdialysis techniques in anesthetized rats. Muscle contraction increased MAP and HR by 37+/-5 mmHg and 23+/-3 bpm, respectively. Extracellular glutamate concentrations, determined using HPLC-ECD, increased from 0.8+/-0.2 to 6.6+/-1.2 ng/5 microliter in the bilateral RVLM areas. Microdialysis of DAME (100 microM) for 30 min attenuated the contraction-evoked increases in MAP, HR, and glutamate levels (20+/-4 mmHg, 10+/-2 bpm, and 1.8+/-0.2 ng/5 microliter, respectively). After microdialysis of naloxone (100 microM) for 30 min into the RVLM, muscle contraction blocked the attenuations (35+/-5 mmHg, 26+/-4 bpm, and 5.8+/-1.0 ng/5 microliter, respectively). Developed muscle tensions were similar throughout the protocol (676+/-38, 678+/-37 and 687+/-37 g, respectively). These results suggest that an opioidergic receptor-mediated mechanism within the RVLM attenuates cardiovascular responses during static exercise via modulating extracellular concentrations of glutamate in the RVLM.

Clinical spectrum and endomyocardial biopsy findings in eosinophilic heart disease

SummaryFourteen cases of heart disease with hypereosinophilia were analyzed employing conventional cardiologic methods, including echocardiography, cardiac catheterization, and endomycoardial biopsy. The cases were divided into four types: (1) Acute carditic (endocarditis, myocarditis, pericarditis; five cases); (2) ventricular dilation (three cases); (3) restrictive (three cases); (4) electric disturbance (three cases). Biopsy revealed significant changes in all cases. In one case of the ventricular dilation type, endomyocardial fibrosis with myocardial degeneration was seen, and in another case mural thrombus formation was shown to be present. In three cases of the restrictive type, endomyocardial fibrosis (EMF) was observed. In two cases of the electric disturbance type, minor right ventricular myocardial degeneration was observed. In two of the three cases of the carditic type and in three of eight cases in other categories, postmyocarditic changes were observed. The course of the disease compared with the type of disorder revealed a short course in the carditic type and a longer course, ranging from 2 to 24 years, with one exception, in the other types. It is also confirmed that the various histopathologic changes can be related to particular clinical presentations. We have shown that the basic changes in eosinophilic heart disease are not restricted to the endomyocardium and that they occur in various parts of the heart causing more widespread manifestations. The more comprehensive term “eosinophilic heart diease” is a preferable description.

Cardiovascular responses and neurotransmitter changes following blockade of nNOS within the ventrolateral medulla during static muscle contraction.

Nitric oxide (NO) is synthesized from L-arginine through the activity of the synthetic enzyme, NO synthase (NOS). Previous studies have demonstrated the roles of the three isoforms of NOS, namely endothelial NOS (eNOS), neuronal NOS (nNOS), and inducible NOS (iNOS) in cardiovascular regulation. However, no investigation has been done to study their individual role in modulating cardiovascular responses during static skeletal muscle contraction. In this study, we determined the effects of microdialyzing a specific nNOS antagonist into the rostral (RVLM) and caudal ventrolateral medulla (CVLM) on cardiovascular responses and glutamatergic/GABAergic neurotransmission during the exercise pressor reflex using rats. We hypothesized that the NO modulation of the exercise pressor reflex was largely influenced by specific nNOS activity within the ventrolateral medulla. Bilateral microdialysis of a selective nNOS antagonist, 1-(2-trifluoromethylphenyl)-imidazole (1.0 microM), for 30 or 60 min into the RVLM potentiated cardiovascular responses and glutamate release during a static muscle contraction. Levels of GABA within the RVLM were decreased. The cardiovascular responses and neurochemical changes to muscle contraction recovered following discontinuation of the drug. In contrast, bilateral application of the nNOS antagonist into CVLM attenuated cardiovascular responses and glutamate release during a static muscle contraction, but augmented GABA release. These results demonstrate that nNOS in the ventrolateral medulla plays an important role in modulating glutamatergic/GABAergic neurotransmission that regulates the exercise pressor reflex, and contributes to the sympathoexcitatory and sympathoinhibitory actions of NO within the RVLM and CVLM, respectively.

Simultaneous glutamate and γ-aminobutyric acid release within ventrolateral medulla during skeletal muscle contraction in intact and barodenervated rats

The purpose of this study was to determine if baroreflex modulates cardiovascular responses and neurotransmitter release within rostral (RVLM) and caudal (CVLM) ventrolateral medulla during static contraction of skeletal muscle using anesthetized rats. We evoked cardiovascular responses by a static muscle contraction and measured simultaneous release of glutamate and gamma-aminobutyric acid (GABA) in both the RVLM and CVLM using microdialysis probes, two inserted bilaterally into the RVLM and two into the CVLM. In intact anesthetized rats, a muscle contraction increased release of glutamate concomitantly in both the RVLM and CVLM along with significant increases in heart rate and arterial blood pressure. In contrast, concentrations of GABA increased within the RVLM, but decreased significantly within the CVLM during the pressor response. These changes were due to contraction-evoked activation of muscle afferents since tibial nerve stimulation following muscle paralysis failed to evoke glutamate, GABA, or any cardiovascular changes. On the other hand, static muscle contractions in baroreceptor denervated rats augmented the increases in heart rate and blood pressure. Furthermore, muscle contraction significantly enhanced the release of glutamate in the RVLM but attenuated its release in the CVLM. In addition, concentrations of GABA within the RVLM were attenuated following a muscle contraction in denervated rats without any changes in GABA within the CVLM. These results demonstrate that the baroreceptors influence cardiovascular responses to static muscle contraction associated with dynamic changes in glutamate and GABA release within the RVLM and CVLM.

Nitric oxide within periaqueductal gray modulates glutamatergic neurotransmission and cardiovascular responses during mechanical and thermal stimuli

We have previously reported that nitric oxide (NO) within the rostral ventrolateral medulla (RVLM) attenuates cardiovascular responses and extracellular concentrations of glutamate during thermal, but not during mechanical nociceptive stimulation (Ishide. T., Maher, T.J., Ally, A. 2003. Role of nitric oxide in the ventrolateral medulla on cardiovascular responses and glutamate neurotransmission during mechanical and thermal stimuli. Pharmacol. Res. 47, 59-68). In this study, we examined the role of nitric oxide within the dorsolateral periaqueductal gray matter (PAG), a higher center integrating nociceptive reflexes, on cardiovascular responses and glutamate release during both mechanical and thermal nociception using anesthetized Sprague-Dawley rats. Two types of stimuli were studied, both activating peripheral A(delta) and C fiber polymodal nociceptors. Noxious mechanical stimulus was given by applying a bilateral hindpaw pinch for 5 s. Mechanical stimulation of a hindlimb increased mean arterial pressure (MAP), heart rate (HR), and extracellular fluid glutamate within PAG by 20+/-3 mmHg, 37+/-6 bpm, and 1.7+/-0.3 ng/5 microl, respectively (n=10). Bilateral microdialysis of L-arginine (1.0 microM), a NO precursor, into the PAG significantly attenuated MAP, HR, and glutamate increases during a mechanical stimulation. Subsequent administration of N(G)-methyl-L-arginine (L-NMMA) (1.0 microM), a NO synthase inhibitor, into the PAG blocked the ability of NO within PAG to modulate the cardiovascular responses to mechanical stimulus. The noxious thermal stimulus was generated by immersing the metatarsus of a hindpaw in water-bath at a temperature of 52 degrees C for 5 s. Similar increases were observed following thermal stimulation: 35+/-5 mmHg, 40+/-6 bpm, and 1.14+/-0.4 ng/5 microl (n=10). L-Arginine attenuated both cardiovascular responses and glutamate increase during thermal nociception. These results demonstrate that NO within the dorsolateral PAG plays a role in modulating cardiovascular responses by altering glutamate concentrations during both thermal and mechanical nociception.

Neurochemistry within ventrolateral medulla and cardiovascular effects during static exercise following eNOS antagonism.

Nitric oxide synthase (NOS), necessary for the production of nitric oxide from l-arginine, exists in three isoforms: neuronal NOS (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS). We have previously demonstrated that blockade of nNOS within the rostral (RVLM) and caudal ventrolateral medulla (CVLM) differentially modulated cardiovascular responses to static exercise [Ishide, T., Nauli, S.M., Maher, T.J., Ally, A., 2003. Cardiovascular responses and neurotransmitter changes following blockade of nNOS within the ventrolateral medulla during static muscle contraction. Brain Res. 977, 80-89]. In this study, we have examined the effects of bilaterally microdialyzing a specific eNOS antagonist into the RVLM and CVLM on cardiovascular responses and glutamatergic/GABAergic neurotransmission during the exercise pressor reflex in anesthetized rats. Bilateral microdialysis of a selective eNOS antagonist, l-N(5)-(1-iminoethyl)ornithine (l-NIO; 10.0 microM) into the RVLM potentiated cardiovascular responses and increased extracellular fluid glutamate levels during a static muscle contraction. At the same time, levels of GABA within the RVLM were decreased. The cardiovascular responses and neurochemical changes to muscle contraction recovered after discontinuation of the drug. In contrast, bilateral application of the eNOS antagonist into the CVLM attenuated cardiovascular responses and glutamate concentrations during a static muscle contraction, but augmented levels of GABA. These results demonstrate that eNOS within the ventrolateral medulla plays an important role in modulating glutamate/GABAergic neurotransmission, that in turn regulates the exercise pressor reflex. The present study provides further evidence of simultaneous sympathoexcitatory and sympathoinhibitory effects of nitric oxide within the RVLM and CVLM involved in the neural control of circulation during static exercise.

Immunohistochemical localization of plasminogen activator inhibitor-1 in human coronary atherosclerotic lesions involved in acute myocardial infarction.

SummaryImmunohistochemical localization of plasminogen activator inhibitor-1 (PAI-1) was studied using the streptavidin-biotin method in human atherosclerotic coronary arteries. The patients (one male and two females), whose ages ranged from 61 to 78 years, died of anteroseptal acute myocardial infarction without having received any thrombolytic therapy. PAI-1 immunoreactivities (IRs) were mainly detected in the endothelial cells, smooth muscle cells, and collagen fibers of the coronary arterial intima and not in thrombi. Remarkable immunohistochemical staining was seen in intimal collagen fibers. In one patient, intimal PAI-1 IRs partly bordered a thrombus and surrounded a large atheroma rich in cholesterol crystals. Our results suggest that PAI-1 is present in both cellular and extracellular components of human coronary atherosclerotic lesions.

Extracellular serotonin changes in VLM during muscle contraction: effects of 5-HT1A-receptor activation.

This study determined whether muscle contraction causes an increase in extracellular levels of serotonin (5-HT) in the rostral (rVLM) or caudal ventrolateral medulla (cVLM) in anesthetized rats. Muscle contraction, evoked by tibial nerve stimulation, increased mean arterial blood pressure (MAP) by 27 +/- 4 mmHg (n = 8). In addition, 5-HT levels in the rVLM were elevated by 65 +/- 9% during the contraction (n = 8). Results were similar over two repeated contractions. In contrast, muscle contraction increased MAP, but not 5-HT, levels in the cVLM (n = 6). Tibial nerve stimulation after muscle paralysis had no effect on either MAP or 5-HT levels in both rVLM and cVLM. Microdialysis of a 5-HT1A agonist, 8-OH-DPAT (10 mM), into the rVLM for 30 min (n = 6) blunted the MAP change and reduced 5-HT release during contraction. Administration of NAN-190, a 5-HT1A antagonist, into the rVLM had no effect on 5-HT release and cardiovascular responses during muscle contraction and blocked the changes in 5-HT, MAP, and heart rate to static contraction after subsequent microdialysis of 8-OH-DPAT. Results demonstrate that 5-HT levels in the rVLM increase during muscle contraction and that 5-HT1A-receptor activation in the rVLM blunts MAP response to muscle contraction via a decrease in the extracellular concentration of 5-HT.This study determined whether muscle contraction causes an increase in extracellular levels of serotonin (5-HT) in the rostral (rVLM) or caudal ventrolateral medulla (cVLM) in anesthetized rats. Muscle contraction, evoked by tibial nerve stimulation, increased mean arterial blood pressure (MAP) by 27 ± 4 mmHg ( n = 8). In addition, 5-HT levels in the rVLM were elevated by 65 ± 9% during the contraction ( n = 8). Results were similar over two repeated contractions. In contrast, muscle contraction increased MAP, but not 5-HT, levels in the cVLM ( n = 6). Tibial nerve stimulation after muscle paralysis had no effect on either MAP or 5-HT levels in both rVLM and cVLM. Microdialysis of a 5-HT1A agonist, 8-OH-DPAT (10 mM), into the rVLM for 30 min ( n = 6) blunted the MAP change and reduced 5-HT release during contraction. Administration of NAN-190, a 5-HT1A antagonist, into the rVLM had no effect on 5-HT release and cardiovascular responses during muscle contraction and blocked the changes in 5-HT, MAP, and heart rate to static contraction after subsequent microdialysis of 8-OH-DPAT. Results demonstrate that 5-HT levels in the rVLM increase during muscle contraction and that 5-HT1A-receptor activation in the rVLM blunts MAP response to muscle contraction via a decrease in the extracellular concentration of 5-HT.

Role of nitric oxide in the ventrolateral medulla on cardiovascular responses and glutamate neurotransmission during mechanical and thermal stimuli

We have previously reported that alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-receptor blockade within the rostral ventrolateral medulla (RVLM) attenuates cardiovascular responses and extracellular concentrations of glutamate during mechanical, but not during thermal stimulation [Pharmacol. Res. 43 (2001) 47]. In this study, we examined the role of nitric oxide (NO) within the RVLM on cardiovascular responses and glutamate release during both mechanical and thermal nociception using anesthetized Sprague-Dawley rats. Two types of stimuli were studied, both activating peripheral Adelta and C fiber polymodal nociceptors. Noxious mechanical stimuli were given by applying a bilateral hindpaw pinch for 5s. The noxious thermal stimuli were generated by immersing the metatarsus of both hindpaws in a water bath at a temperature of 52 degrees C for 5s. Mechanical stimulation of both hindlimb extremities increased mean arterial pressure (MAP), heart rate (HR), and extracellular fluid glutamate by 14+/-2 mmHg, 35+/-5 bpm, and 1.4+/-0.3 ng/5 microl, respectively (n=8). Similar responses were observed following thermal stimulation: 40+/-4 mmHg, 44+/-6 bpm, and 0.97+/-0.2 ng/5 microl (n=8). Bilateral microdialysis of L-arginine (1.0 microM), a nitric oxide precursor, into the RVLM had no effects on MAP, HR, and glutamate increases during mechanical stimulation. However, L-arginine attenuated these responses during thermal nociception. Subsequent administration of L-NMMA (1.0 microM), a NOS inhibitor, reversed the attenuations. These results show that nitric oxide most likely plays a role in modulating cardiovascular responses by altering glutamate concentrations within the RVLM during thermal but not mechanical nociception. Overall, the present study delineates the differential central integrative mechanisms that regulate processing of sensory impulses arising from peripheral stimulation.