Biomedical subjects

M Kumada

Publications and source records attributed to M Kumada.

At least 37 records · Page 2Linked to original sources

PubMed1999

Responses of blood pressure and catecholamine metabolism to high salt loading in endothelin-1 knockout mice.

The molecular mechanism responsible for salt sensitivity is poorly understood. Mice heterozygous for the null mutation of the endothelin-1 (ET-1) gene, Edn1, may be a potential tool for studying this mechanism, because they have elevated blood pressure and disturbances in central sympathetic nerve regulation. In the present study, we used this mouse model to examine the degree to which ET-1 contributes to the responses of blood pressure and catecholamine metabolism to high salt loading. Male Edn1+/- heterozygous mice and Edn1+/+ wild-type littermates were given either a high salt (8%) or a normal salt (0.7%) diet for 4 wk. During the normal diet, renal ET-1 levels in Edn1+/- mice were approximately 50% lower than ET-1 levels in wild-type mice, whereas the high salt diet decreased renal ET-1 levels by about 50% in both Edn1+/- and wild-type mice. The high salt diet significantly increased urinary sodium excretion and fractional excretion of sodium (FENa) but did not affect circulating plasma volume, serum electrolytes, creatinine clearance, or systemic blood pressure. In addition, urinary norepinephrine and normetanephrine excretion were significantly increased, indicating that salt loading can increase sympathetic nerve activity in normal mice. These responses to salt loading did not differ between Edn1+/- mice and their wild-type littermates. We conclude that physiological changes in ET-1 production do not affect the responses of blood pressure and catecholamine metabolism to salt loading, although the renal ET-1 content is decreased by salt loading.

Animals↗

PubMed1998

Renal sympathetic nerve activity in mice: comparison between mice and rats and between normal and endothelin-1 deficient mice.

Recently generated knockout mice with disrupted genes encoding endothelin (ET)-1 showed an elevation of arterial blood pressure (AP) and supplied an evidence for intrinsic ET-1 as one of the physiological regulators of systemic AP. Little is yet known, however, why deficiency of ET-1, which was originally found as a potent vasoconstrictor, led to higher AP in these mice. To address this apparent paradox, we first developed a method to measure renal sympathetic nerve activity (RSNA) in mice using rats as reference and successively compared it between normal and ET-1 deficient mice. RSNA was successfully recorded in urethane-anesthetized and artificially ventilated mice by a slight modification of the method used for rats. At basal condition, mean AP (MAP) and RSNA in ET-1 deficient mice (105+/-2 mmHg and 9.71+/-1.49 muVs, n=20) were significantly higher than those in wild-type mice (96+/-2 mmHg and 5. 07+/-0.70 muVs, n=25). Basal heart rate (HR) and baroreflex-control of HR was not significantly different between the two. On the other hand, resting RSNA, RSNA range, and maximum RSNA were significantly greater in ET-1 deficient mice, and thus MAP-RSNA relationship was upwards reset. Hypoxia-induced increase in RSNA was not different between ET-1 deficient (73.4+/-9.4%) and wild-type mice (91.2+/-12.0%), while hypercapnia-induced one was significantly attenuated in ET-1 deficient mice (18.8+/-3.6 vs. 39.1+/-5.2% at 10% CO2). These results indicate that endogenous ET-1 participates in the central chemoreception of CO2 and reflex control of the RSNA. Baroreceptor resetting and normally preserved hypoxia-induced chemoreflex may explain a part of the elevation of AP in ET-1 deficient mice.

Animals↗

PubMed1998

Impaired anaphylactic responses with intact sensitivity to endotoxin in mice lacking a platelet-activating factor receptor.

Platelet-activating factor (PAF) is a potent phospholipid mediator with diverse biological activities in addition to its well-known ability to stimulate platelet aggregation. Pharmacologic studies had suggested a role for PAF in pregnancy, neuronal cell migration, anaphylaxis, and endotoxic shock. Here we show that disruption of the PAF receptor gene in mice caused a marked reduction in systemic anaphylactic symptoms. Unexpectedly, however, the PAF receptor-deficient mice developed normally, were fertile, and remained sensitive to bacterial endotoxin. These mutant mice clearly show that PAF plays a dominant role in eliciting anaphylaxis, but that it is not essential for reproduction, brain development, or endotoxic shock.

Anaphylaxis↗

PubMed1998

Design of a compact synchrotron light source for medical applications at NIRS.

A synchrotron light source dedicated to medical applications is required to be compact for installation in limited spaces at hospitals. The NIRS storage ring, with a circumference of 44.8 m, is designed to accelerate electrons up to 1.8 GeV and to store a beam of 400 mA. The ring is composed of superconducting bending magnets for downsizing. A beam of 300 MeV is injected into the ring from a microtron operated at an L-band RF frequency. There are two superconducting multipole wigglers with nine poles and a maximum field of 8 T, which can produce a photon flux of about 1.4 x 10(13) photons s(-1) mrad(-1) (0.1% bandwidth)(-1) at 33 keV used for coronary angiography.

Journal Article↗

PubMed1998

Systemic and renal response to salt loading in endothelin-1 knockout mice.

Endothelin-1 (ET-1) knockout mice demonstrate elevated blood pressure, which may be associated with disturbance in central cardiorespiratory regulation. In this study we examined responses to salt loading in ET-1 knockout mice to investigate whether ET-1 is involved in the pathophysiology of salt-sensitive hypertension. Male Edn1+/- heterozygous mice and their wild-type littermates were fed either a high NaCl (8%) or a normal (0.2%) diet for 4 weeks. Systemic blood pressure and tissue ET-1 levels were measured as well as several parameters relating to sodium handling and volume homeostasis. On normal diet, renal ET-1 levels of Edn1+/- mice were about 50% of those of wild-type mice. A high-salt diet caused a significant decrease in renal ET-1 levels by about 50% in both groups. Urine volume, urinary sodium excretion, and FENa in mice on the 8% NaCl diet were significantly higher than those in mice on the 0.2% NaCl diet, whereas there were no differences in circulating plasma volume, serum electrocytes, and creatinine clearance. These responses were similar in Edn1+/- and wild-type mice. Although systemic blood pressure was significantly higher in Edn1+/- mice than in the wild-type, the effect of salt loading on blood pressure was not significant in either Edn1+/- or wild-type mice. We conclude that changes in ET-1 production within a physiologic range do not affect salt sensitivity, although renal ET-1 content is decreased by salt loading.

Animals↗

PubMed1998

Cranial and cardiac neural crest defects in endothelin-A receptor-deficient mice.

Neural crest cells arise in the dorsal aspect of the neural tube and migrate extensively to differentiate into a variety of neural and non-neural tissues. While interactions between neural crest cells and their local environments are required for the proper development of these tissues, little information is available about the molecular nature of the cell-cell interactions in cephalic neural crest development. Here we demonstrate that mice deficient for one type of endothelin receptor, ETA, mimic the human conditions collectively termed CATCH 22 or velocardiofacial syndrome, which include severe craniofacial deformities and defects in the cardiovascular outflow tract. We show that ETA receptor mRNA is expressed by the neural crest-derived ectomesenchymal cells of pharyngeal arches and cardiac outflow tissues, whereas ET-1 ligand mRNA is expressed by arch epithelium, paraxial mesoderm-derived arch core and the arch vessel endothelium. This suggests that paracrine interaction between neural crest-derived cells and both ectoderm and mesoderm is essential in forming the skeleton and connective tissue of the head. Further, we find that pharyngeal arch expression of goosecoid is absent in ETA receptor-deficient mice, placing the transcription factor as one of the possible downstream signals triggered by activation of the ETA receptor. These observations define a novel genetic pathway for inductive communication between cephalic neural crest cells and their environmental counterparts.

Animals↗

PubMed1998

Oxidative stress increases adrenomedullin mRNA levels in cultured rat vascular smooth muscle cells.

We investigated the effect of oxidative stress on the expression of adrenomedullin (AM) mRNA in cultured rat vascular smooth muscle cells (VSMCs), using diethyldithiocarbamate (DDC), which is known to inhibit endogenous Cu, Zn superoxide dismutase (SOD) and to increase superoxide (O2-). DDC (10 mM) increased O2- levels produced from rat VSMCs in a time-dependent fashion. Concomitantly, DDC increased the expression of AM mRNA for up to 24 h, although it did not affect the expression of beta-actin mRNA. Thus, oxidative stress enhanced AM production in rat VSMCs, which may compensate for oxidative-stress-induced vasoconstriction.

Actins↗

PubMed1998

Excitation of baroreceptors depresses A- and C-components of the somato-cardiac sympathetic reflex in anesthetized rats.

The effect of baroreceptor activation on somato-cardiac sympathetic reflex discharges was examined in urethane-anesthetized, vagotomized, and artificially ventilated rats. Single shock stimulation of myelinated (A) and unmyelinated (C) fibers in the tibial nerve of the left hindlimb elicited two separate excitatory reflex discharge components in a branch of the cardiac sympathetic nerve. They are termed the A- and C-components of the somato-cardiac sympathetic reflex discharges. When aortic nerves (AN) and carotid sinus nerves (CSN) were intact, a sudden increase in mean arterial blood pressure to about 150 mmHg induced by I.V. injection of phenylephrine (50 micro/kg) depressed the A- and C-components by up to 47 +/- 5.4 and 37 +/- 7.7% of the control values, respectively. However, bilateral sino-aortic denervation completely abolished the pressure-induced depression of both components. We conclude that baroreceptor afferent signals from the AN and CSN inhibit both A- and C-components of the excitatory somato-cardiac sympathetic reflex discharges. This and other previous evidence mentioned in the text indicate that inhibitory cardiac sympathetic reflexes originating from arterial baroreceptors and excitatory ones originating from somatic afferents interact, probably at the brainstem.

Anesthetics, Intravenous↗

PubMed1997

Physiological role of brain endothelin in the central autonomic control: from neuron to knockout mouse.

Although endothelin (ET) was discovered as a potent vascular endothelium-derived constricting peptide, its presumed physiological and pathophysiological roles are now considered much more diverse than originally though. Endothelin in the brain is thought to be deeply involved in the central autonomic control and consequent cardiorespiratory homeostasis, possibly as a neuromodulator or a hormone that functions locally in an autocrine/paracrine manner or widely through delivery by the cerebrospinal fluid (CSF). This notion is based on the following lines of evidence. (1) Mature ET, its precursors, converting enzymes, and receptors all are detected at strategic sites in the central nervous system (CNS), especially those controlling the autonomic functions. (2) The ET is present in the CSF at concentrations higher than in the plasma. (3) There is a topographical correspondence of ET and its receptors in the CNS. (4) The ET is released by primary cultures of hypothalamic neurons. (5) When ET binds to its receptors, intracellular calcium channels. (6) An intracerebroventricular or topical application of ET to CNS sites elicits a pattern of cardiorespiratory changes accompanied by responses of vasomotor and respiratory neurons. (7) Recently generated knockout mice with disrupted genes encoding ET-1 exhibited, along with malformations in a subset of the tissues of neural crest cell lineage, cardiorespiratory abnormalities including elevation of arterial pressure, sympathetic overactivity, and impairment of the respiratory reflex. Definitive evidence is expected from thorough analyses of knockout mice by applying conventional experimental methods.

Amino Acid Sequence↗

PubMed1997

Differential central modulation of the baroreflex by salt loading in normotensive and spontaneously hypertensive rats.

In salt-sensitive hypertensive animal models and human subjects compared with their salt-resistant counterparts, sympathetic activity is abnormally enhanced during a high salt diet. We examined whether salt loading differentially modulates the arterial baroreceptor reflex (ABR), a major control mechanism of arterial pressure and sympathetic vasomotor activity, in young normotensive Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). Six-week-old WKY and SHR were fed a normal (0.66%) or high (8.00%) salt diet for 4 weeks. After the diet regimen, baseline levels of mean arterial pressure (MAP), renal sympathetic nerve activity (RSNA), and the overall and central properties of the ABR were compared among the four groups of rats under halothane anesthesia. In WKY, a high salt diet did not affect baseline arterial pressure and RSNA but potentiated the ABR, as evidenced by an increase in the maximal slope of MAP-RSNA and MAP-heart rate relationships. In SHR, by contrast, salt loading accelerated hypertension and sympathetic overactivity and impaired the ABR. Salt-induced modulation of the ABR was associated with that of the central property, since reflex inhibition of RSNA by stimulation of the aortic depressor nerve was augmented in WKY and attenuated in SHR. These results suggest that differential modulation of the central mechanism subserving the baroreflex control of sympathetic activity at least partly accounts for the difference in salt sensitivity between WKY and SHR.

Animals↗

PubMed1997

Determination of ventilatory volume in mice by whole body plethysmography.

We examined the reliability of whole-body plethysmography (WBP) for determining the ventilatory volume in mice. The degree of agreement of tidal volume (VT) determined consecutively by WBP with that by direct plethysmography (DP) or pneumotachography (PT) was analyzed using urethane-anesthetized young male 129/Sv mice. Between WBP and DP, the mean difference of VT was 0.000 ml with a 95% confidence interval of -0.003 to 0.002, whereas that between WBP and PT was 0.000 ml (-0.004 to 0.003). The limits of agreement, or mean +/- 2SD of the difference, were -0.008 and 0.008 ml in WBP vs. DP, and -0.011 and 0.010 ml in WBP vs. PT. These values were 5.1-6.6% of the average VT (0.157 ml in WBP and DP, and 0.165 ml in WBP and PT). Thus, VT can be determined by WBP without any bias and with a systemic error of less than 7% of that measured by the other direct methods examined. Through applying WBP to conscious 129/Sv and 129/SvJ-ICR hybrid mice, we further studied the effects of differences in mouse strain, age and sex on respiratory rate (f) and minute volume (V = VT.f), and their reflex changes in responses to systemic hypoxia and hypercapnia. V was significantly greater in young (10-16 weeks old) male 129/Sv than in young male 129/SvJ-ICR hybrids (139 +/- 11 and 73 +/- 8 ml/min/100 g, respectively), while the pattern of reflex responses was similar. In aged (32-44 weeks old) male 129/Sv, the basal level of V and its reflex increases were all diminished as compared to those in their young counter parts. However, we were unable to find significant differences between aged male and female 129/Sv mice. In conclusion, WBP is a useful and reliable method to determine ventilatory parameters in conscious and anesthetized mice. Furthermore, there are considerable strain- and age-related differences in baseline ventilation and reflex responses to hypoxia and hypercapnia.

Age Factors↗

PubMed1997

[Central autonomic control in human hypertension].

Sympathetic nerve fibers innervating the heart and blood vessels exhibit tonic activity which originates from vasomotor neurons in the rostral ventrolateral medulla (RVLM). Whether sympathetic nerve activity (SNA) is elevated in humans with essential hypertension is still unsettled. Recording of muscle SNA (MSNA) by the microneurographic method mostly points to elevated SNA in hypertensives including borderline and mildly hypertensive patients. Exaggerated MSNA is most conspicuous in the accelerated hypertensives. Power spectral analyses of heart rate and blood pressure variability provide controversial results on this issue. Nevertheless, most studies demonstrate altered sympathetic cardiovascular control in hypertensive subjects. In young borderline hypertensives, the sympathetic response to mental stress or passive tilting is exaggerated, whereas in elderly subjects it is attenuated.

Blood Pressure↗

PubMed1996

Protein kinase C inhibits the CAK-CDK2 cyclin-dependent kinase cascade and G1/S cell cycle progression in human diploid fibroblasts.

Serum stimulation of human diploid fibroblast IMR-90 cells leads to phosphorylation of p33CDK2 at Thr160 and activation of CDK2 kinase, a necessary event for G1/S transition. We report that serum stimulation causes a gradual, sustained increase in the activity of CDK-activating kinase (CAK) that phosphorylates CDK2 at Thr160, which starts by 5 h after serum stimulation and reaches the maximal plateau level at around the G1/S boundary. In this cell type addition of phorbol-12, 13-dibutyrate 5 h but not 16 h after serum stimulation completely inhibits CDK2 kinase activation and DNA synthesis. Phorbol ester treatment does not reduce the protein level of p33CDK2, but does inhibit serum-stimulated increases in the CAK activity and CDK2 phosphorylation at Thr160. The suppression of the CAK activity by the phorbol ester is accompanied by decreases in the message levels of both CDK7 and cyclin H, the catalytic and the positive regulatory subunit of CAK, respectively. These results indicate that in IMR-90 cells activation of protein kinase C in the late G1 phase causes cell cycle arrest before the G1/S boundary at least in part through downregulation of CAK and CAK-mediated CDK2 phosphorylation and activation.

Antibodies↗

PubMed1996

Impaired ventilatory responses to hypoxia and hypercapnia in mutant mice deficient in endothelin-1.

We studied respiratory functions in mutant mice deficient in endothelin-1 (ET-1) generated by gene targeting. In conscious adult mice heterozygous for ET-1 gene mutation (ET+/- heterozygous mice), arterial PO2 was significantly lower, PCO2 tended to be higher, and pH tended to be lower than in wild-type littermates. When these conscious mice breathed room air, respiratory minute volume and rate, determined by body plethysmography, were not significantly different between the two groups. However, when ET+/- heterozygous mice were subjected to systemic hypoxia (1:1 air-N2) or hypercapnia (5% CO2-95% O2), increases in respiratory minute volume were significantly attenuated. In conscious newborn ET-/- homozygous mice delivered by cesarean section and tracheotomized, ventilatory responses to systemic hypoxia and hypercapnia, regularly observed in newborn wild-type mice, were almost totally absent. In urethan-anesthetized adult ET+/- heterozygous mice, increases in phrenic nerve discharges in response to hypoxia and hypercapnia were significantly attenuated. Our results demonstrate that ventilatory responses to hypoxia and hypercapnia are impaired in ET-1-deficient mice and suggest that endogenous ET-1 participates in the physiological control of ventilation.

Animals↗

PubMed1995

Molecular cloning and functional analysis of a novel P2 nucleotide receptor.

The cDNA encoding a novel P2 receptor was isolated from rat aortic smooth muscle cell library and functionally characterized. The cloned P2 receptor exhibits structural features characteristic of the G protein-coupled receptor family and shows 44 and 38% amino acid identity with previously cloned rat P2U and chicken P2Y receptors, respectively. The cloned P2 receptor is functionally coupled to phospholipase C but not to adenylate cyclase in C6 rat glioma cells transfected with the cloned P2 expression vector. The rank order of agonist potency as judged by intracellular Ca2+ mobilization responses is UTP > ADP = 2-methylthioATP > ADP beta S > ATP = ATP gamma S, which is not compatible with any of the previously characterized P2 receptor subtypes. The nonselective P2 antagonists, suramin and reactive blue-2, inhibit nucleotide-induced phospholipase C activation in cells expressing the cloned P2 receptor. The cloned P2 receptor mRNA is abundantly expressed in various rat tissues including lung, stomach, intestine, spleen, mesentery, heart, and, most prominently, aorta. The results indicate that the novel metabotropic P2 receptor has pharmacological characteristics distinct from any of P2 receptor subtypes thus far identified and suggest the existence of a novel regulatory system by extracellular nucleotides of potential significance.

Adenine Nucleotides↗

PubMed1995

Involvement of intact inositol-1,4,5-trisphosphate-sensitive Ca2+ stores in cell cycle progression at the G1/S boundary in serum-stimulated human fibroblasts.

Thapsigargin, a selective inhibitor of the endoplasmic reticulum Ca2+ pump, has been shown to deplete inositol-1,4,5-trisphosphate-sensitive Ca2+ stores. Here we report that when thapsigargin was introduced to serum-stimulated human fibroblasts at a time point just before the G1/S boundary, it completely inhibited expression of cyclin A, activation of p33CDK2 cyclin-dependent kinase and initiation of DNA synthesis. In contrast, the Ca2+ mobilizing ionophore ionomycin was without effect. These findings indicate that Ca2+ inside the inositol-1,4,5-trisphosphate-sensitive Ca2+ stores plays a pivotal role for traverse across the G1/S transition point.

CDC2-CDC28 Kinases↗

PubMed1995

Impaired development of the thyroid and thymus in endothelin-1 knockout mice.

We have previously demonstrated that endothelin-1 (ET-1) is essential to the development of neural crest-derived craniofacial and cardiovascular structures. In this study we evaluated the intrauterine growth and development of the thyroid and thymus glands in Edn1-/- homozygous mice. Edn1-/- homozygous mice were smaller than their heterozygous or wild-type littermates (about 90% of normal body weight). The thyroid and thymus of Edn1-/- homozygous mice were smaller than those of normal mice and were not fused in the midline, resulting in two separate lobules. The thymus of Edn1-/- homozygous mice is not descended into the normal position. These results suggest that ET-1 is important for normal development of the glands in the neck region, as well as for systemic growth. The combination of abnormalities in Edn1-/- homozygous mice suggests that these mice might serve as an animal model for the human diseases DiGeorge syndrome and velocardiofacial syndrome.

Animals↗

PubMed1995

Gene expression of endothelin isoforms and receptors in endothelin-1 knockout mice.

Gene expression of endothelin (ET) isoforms and receptors was examined in ET-1 knockout mice and compared with that of wild-type mice. ET-1 mRNA and peptide levels were in parallel with the number of Edn1 authentic alleles. ET-2 mRNA levels in intestine and ET-3 mRNA and/or peptide levels in lung, brain, and intestine were not different among Edn1+/+ wild-type, Edn1+/- heterozygous, and Edn1-/- homozygous mice. ET(A) and ETB receptor mRNA levels in several organs also were not different among Edn1+/+ wild-type, Edn1+/- heterozygous, and Edn1-/- homozygous mice. These results suggest that gene expression of ET-2, ET-3, and ET(A/B) receptors is not affected by levels of ET-1 production and is regulated independently. Neither compensation by redundant expression of other ET isoforms nor upregulation of ET receptors could explain the elevated blood pressure in Edn1+/- heterozygous mice.

Animals↗