Akihiko Himeno

Nagasaki University Hospital, Nagasaki, Nagasaki, Japan

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Publications (41)97.35 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: 1. Interaction in the recognition of endothelin-1 (ET-1), a typical bivalent ET receptor-ligand, between ETA and ETB receptors was investigated in the rat anterior pituitary gland, using our quantitative receptor autoradiographic method with tissue sections preserving the cell-membrane structure and ET receptor-related compounds. 2. In saturation binding studies with increasing concentrations (0.77-200 pM) of 125I-ET-1 (nonselective bivalent radioligand), 125I-ET-1 binding to the rat anterior pituitary gland was saturable and single with a KD of 71 pM and a Bmax of 120 fmol mg(-1). When 1.0 microM BQ-123 (ETA antagonist) was added to the incubation buffer, binding parameters were 8.3 pM of KD and 8.0 fmol mg(-1) of Bmax, whereas 10 nM sarafotoxin S6c (ETB agonist) exerted little change in these binding parameters (KD, 72 pM; Bmax, 110 fmol mg(-1)). 3. Competition binding studies with a fixed amount (3.8 pM) of 125 I-ET-1 revealed that when 1.0 microM BQ-123 was present in the incubation buffer, ETB receptor-related compounds such as sarafotoxin S6c, ET-3, IRL1620 (ETB agonist), and BQ-788 (ETB antagonist) competitively inhibited 125I-ET-1 binding with K(i)s of 140,18,350 pM, and 14 nM, respectively, however, these compounds were not significant competitors for 125I-ET-1 binding in the case of absence of BQ-123. 4. In cold-ligand saturation studies with a fixed amount (390 pM) of 125I-IRL 1620 (ETB radioligand), IRL1620 bound to a single population of the ETB receptor, and no change was observed in binding characteristics in the presence of 1.0 microM BQ-123. 125I-IRL1620 binding was competitively inhibited by ET-1 and ET-3 in the absence of BQ-123, with K(i)s of 20 and 29 pM, respectively, the affinities being much the same as those of 29 nM, in the presence of 1.0 microM BQ-123. 5. Two nonbivalent ETA antagonists, BQ-123 and PD151242, were highly sensitive and full competitors for 125I-ET-1 binding (5.0 pM), in the presence of 10 nM sarafotoxin S6c. 6. Taken together with the present finding that mRNAs encoding the rat ETA and the ETB receptors are expressed in the anterior pituitary gland, we tentatively conclude that although there are ETA and ETB receptors with a functional binding capability for ET receptor-ligands, the ETB receptor does not independently recognize ET-1 without the aid of the ETA receptor. If this thesis is tenable, then ET-1 can bridge between the two receptors to form an ETA-ETB receptor heterodimer.
    Cellular and Molecular Neurobiology 05/2002; 22(2):207-26. DOI:10.1023/A:1019822107048 · 2.51 Impact Factor
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    ABSTRACT: 1. In situ hybridization done using a 35S-cRNA probe was carried out to obtain information on the expressions of the SA gene in brains and kidneys of the spontaneously hypertensive rat (SHR) strain obtained from the Izumo colony (/Izm) and from Charles River Laboratories (/Crj). 2. In the brain, SA mRNA expression was most abundantly observed in epithelial cells of the choroid plexus. High to moderate levels was present on neurons of the CA1-CA4 pyramidal cell layer and the dentate gyrus of the hippocampus and the cerebellar Purkinje cell layer. The solitary tract nucleus and the dorsal motor nucleus of the vagus expressed the SA gene at very low levels. An increase in the expression was noted in the choroid plexus of WKY/Crj; there was no difference, however, in expression levels of other brain areas between WKY/Izm, SHR/Izm, and SHRSP/Izm, and between WKY/Crj and SHR/Crj. 3. In the kidney, expression signals of SA mRNA were observed in renal medullary rays and focal cortex of WKY/Izm, SHR/Izm, SHRSP/Izm, and SHR/Crj, whereas mRNA expression in the WKY/Crj kidney was observed in medullary rays and outer strips of the outer medulla. Microscopically, hybridization signals were predominant in the proximal tubules. 4. Expression densities decreased only in the kidney of WKY/Crj in 4-and 8-week-old rats, but not in the WKY/Izm kidney, compared with findings in SHR and SHRSP kidneys. These observations are in good agreement with data from Northern blot analysis. 5. The SA gene expressions in the brain and the kidney seem not to relate to states of elevated blood pressure, but rather to strain differences. Abundant expressions in the brain and the kidney may mean that the SA gene plays a role in the water-electrolyte transport system. It is noteworthy that there are neuronal expressions of the SA gene in hippocampal pyramidal cells and cerebellar Purkinje cells.
    Cellular and Molecular Neurobiology 01/2001; 20(6):633-52. DOI:10.1023/A:1007042506936 · 2.51 Impact Factor
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    ABSTRACT: 1. We examined time- and cell-type-dependent changes in endothelin (ET)-1-like immunoreactivity, ET receptors binding and nitric oxide (NO) synthase (NOS) activity in CA1 subfields of the hippocampus of stroke-prone spontaneously hypertensive rats subjected to a 10-min bilateral carotid occlusion and reperfusion.2. Microglia aggregated in accord with neuronal death and expressed a high density of ETB receptors and an intense NOS activity in the damaged CA1 pyramidal cell layer, 7 days after the induced transient forebrain ischemia. The increased NOS activity and ETB receptor in microglia disappeared 28 days after this transient ischemia.3. In contrast to microglia, astrocytes presented a moderate level of ET-1-like immunoreactivity, ETB receptors, and NOS activity in all areas of the damaged CA1 subfields, 7 days after the ischemia. These events were further enhanced 28 days after the ischemia.4. In light of these findings, the possibility that the microglial and the astrocytic ETB/NO system largely contributes to development of the neuronal death and to reconstitution of the damaged neuronal tissue, respectively, in the hippocampus subjected to a transient forebrain ischemia would have to be considered.
    Cellular and Molecular Neurobiology 09/2000; 20(5):541-551. DOI:10.1023/A:1007007710703 · 2.51 Impact Factor
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    ABSTRACT: Endothelins are potent vasoactive peptides that bind to their specific receptors, playing an important role in the CNS under physiological and pathophysiological conditions. Astrocytes, which have been shown to express these receptors, also have a considerable role to play under physiological and pathophysiological conditions, particularly those involved in delayed neuronal death. We carried out in vitro receptor autoradiographic binding experiments using specific ligands for endothelin receptors on cultured rat astrocytes. On astrocytes, the specific binding sites for (125)I-PD151242 (a selective endothelin A receptor antagonist) and (125)I-IRL 1620 (a selective endothelin B receptor agonist) were detected. We also characterized the qualitative and quantitative changes of endothelin receptors 24 h after subjecting cultured rat astrocytes to a transient 4-h hypoxia/hypoglycemia insult, used as a model of delayed neuronal death. After transient hypoxia/hypoglycemia, the number of endothelin B receptors increased significantly on cultured astrocytes, but this did not occur among the endothelin A receptors. These findings suggest that the astrocytic effects associated with endothelin in delayed neuronal death include gliosis or the repair process or both, manifested primarily by an increase in the number of endothelin B receptors. This rise does not require interaction with other types of CNS cells. Endothelin A receptors might have a role taking their number into consideration on rat astrocytes under both physiological and pathophysiological conditions.
    Glia 08/2000; 31(1):91-4. DOI:10.1002/(SICI)1098-1136(200007)31:13.0.CO;2-E · 6.03 Impact Factor
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    ABSTRACT: Endothelins (ETs) work as growth factors for fibroblastic growth. Keloids are clinically unpleasant scars within the skin that grow beyond the confines of the original wound. We compared the expression of ET receptors between human skins with hypertrophic scars and keloids, by using our quantitative receptor imaging system. The ET receptor binding sites were distributed in skin areas anatomically corresponding to the epidermis, the superficial and deep dermis, and the vascular bed. The ETA receptor was apparently observed in the skin areas, whereas the ETB receptor was only slightly detected, as deduced from the finding that mRNA encoding the human ETA receptor is expressed in the fibroblast and the vascular bed. The ETA receptor densities of keloid and highly hypertrophic scar were significantly higher than those of normal skin and moderately and mildly hypertrophic scar. There seemed to be no differences in the density between keloid and highly hypertrophic scar. In a keloid skin we noted a possible existence of atypical vascular bed ETA receptor with a low affinity to ETA antagonist of PD151242 and FR139317. The ETA receptor of endogenous growth factors possibly play an etiological role in dysfunctions of the fibroblast in hypertrophic scars, and a typical ETA receptor may be related to the growth of keloid
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    ABSTRACT: We used the quantitative receptor autoradiographic method with a radioligand of [ 125I ]lysergic acid diethylamide ([125I]LSD) to quantitate platelet serotonin (5-HT)2A receptors in affective disorders. Specific binding of [125I]LSD to human platelet pellet sections was saturable, and of high affinity and single. Both ketanserin and spiperone, 5-HT2A selective ligands, inhibited [125I]LSD binding to human platelet pellets with high potency (IC50 values of 0.15 and 0.19 nM, respectively), whereas 5-HT and paroxetine, selective 5- HT re-uptake inhibitors, inhibited binding with a very low potency. These data confirmed that binding sites of human platelet pellets specifically labelled by [125I] LSD were 5-HT2A receptors. The number of 5-HT2A receptors (Bmax of [ 125I] LSD binding) of human platelets obtained from drug-free depressed patients was significantly higher than those of healthy volunteers. There were no statistical differences in the number of 5-HT2A receptors between depressed patients with and without suicidal behaviors. The increased number in platelet 5-HT2A receptotrs may indicate a hyperfunction of the central 5-HT2A receptors. The method with human platelets pellet sections we used is simple and sensitive for investigating platelet 5-HT2A receptors, a diagnostic and therapeutic marker in depressive disorders, in the clinical research
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    ABSTRACT: In the gerbil brain, most of the []Sarcosine1–Angiotensin II binding sites are atypical, not sensitive to displacement with selective Angiotensin II AT1 and AT2 receptor ligands. A similar atypical binding profile exists in the gerbil kidney, where binding is highly expressed. We isolated a 2197 base pair clone from a gerbil kidney cDNA library which encodes a 359 amino acid protein with higher than 90% homology to other mammalian angiotensin II AT1 receptors. When expressed in COS-7 cells, stimulation by Angiotensin II of both the cloned gerbil receptor or the human AT1 receptor enhanced IP3 production to a similar degree. In COS-7 cells, the gerbil receptor also had a ligand affinity profile similar to that of the human AT1 receptor, but showed greatly reduced affinity for losartan (IC50=3480±174 nM). In the gerbil brain, in situ hybridization revealed receptor mRNA in circumventricular organs, selective hypothalamic, midbrain and brain stem areas, and in the hippocampus, where high mRNA expression was detected in the stratum pyramidale of the CA1 and CA2 subfields, and in the stratum granulosum of the dentate gyrus. The expression pattern of receptor mRNA corresponded well with that of atypical []Sar1–Ang II binding. In situ hybridization and Southern blot experiments using riboprobes against the open reading frame and the 3′-untranslated region of the cloned gerbil Ang II receptor cDNA suggest that gerbils have, like other rodents, two AT1 receptor subtypes. The receptor mRNA distribution of the cloned gerbil Ang II receptor corresponds to the distribution of AT1A receptors described in other rodent species.
    Molecular Brain Research 11/1998; 60(2-60):234-246. DOI:10.1016/S0169-328X(98)00187-9 · 2.00 Impact Factor
  • A Himeno · K Shigematsu · T Taguchi · M Niwa ·
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    ABSTRACT: 1. 125I-Endothelin (ET)-1 binding to the rat anterior pituitary gland was saturable and single, with a Kd of 71 pM and a Bmax of 120 fmol/mg. 2. When 1.0 microM BQ-123 (ETA antagonist) was added to the incubation buffer, the binding parameters were 8.3 pM and 8.0 fmol/mg, whereas 10 nM sarafotoxin S6c (ETB agonist) exerted little change in these binding parameters (Kd, 72 pM; Bmax, 110 fmol/mg). 3. ETB receptor-related compounds such as sarafotoxin S6c, ET-3, IRL1620, and BQ-788 competitively inhibited 125I-ET-1 binding, only when 1.0 microM BQ-123 was present in the incubation buffer. 4. Thus, the ETB receptor is capable of binding ET-1 when the ETA receptor is being occupied by BQ-123. A collaboration mechanism between the ETA and the ETB receptor may function in the recognition of ET-1, a typical "bivalent" ligand.
    Cellular and Molecular Neurobiology 09/1998; 18(4):447-52. DOI:10.1023/A:1022557717481 · 2.51 Impact Factor
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    ABSTRACT: We studied the characterization of receptor-mediated G protein activity by nociceptin throughout brain regions, using in situ GTPgammaS binding autoradiography. Nociceptin-stimulated GTPgammaS binding was markedly observed in amygdala, hippocampal pyramidal cell layers, temporal and entorhinal cortex, infralimbic organ, anterior olfactory nucleus, and rostral part of thalamus. These nociceptin-stimulated activities were not affected by naloxone, naltrindol nor norbinaltorphimine which completely blocked mu-, delta- or kappa-opioid agonist-stimulated GTPgammaS binding, respectively. In addition, the distribution of nociceptin-stimulated activities throughout brain regions was found to be different from such opioid receptor-mediated ones.
    Neuroscience Letters 12/1997; 237(2-3):113-6. DOI:10.1016/S0304-3940(97)00807-0 · 2.03 Impact Factor
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    ABSTRACT: Seven days after an intracerebroventricular injection of 0.8 microgram kainic acid, a time of neural tissue-repair after damage, we applied our receptor autoradiographic method to examine changes in the endothelin receptors in kainic acid-induced neural lesions of the rat brain. There were belt-shaped areas with the de novo expressed [125I]endothelin-1 binding sites in the damaged hippocampus CA1, CA3, and CA4 subfields. We also noted a homogeneous zone with a low binding-density, the area sandwiched by the belt-shaped areas. In a "remote" area corresponding anatomically to the deep soma layer of the piriform cortex plus lateral parts of amygdaloid complex we noted a well-defined area with "punched hole-figure" of low density [125I]endothelin-1 binding sites. The lesion was surrounded by areas rich in binding sites. The de novo expressed [125I]endothelin-1 binding sites were characterized endothelin B receptor. Microglia were present in the area with "punched hole-figure" and in the hippocampus pyramidal cell layer with neuronal death. In contrast to microglia, astrocytes were rich with hypertrophia in kainic acid-induced neural lesions anatomically corresponding to areas with the de novo endothelin B receptor. Taken together with the present observations of microscopic evidence of cellular distribution, we suggest that the de novo expressed endothelin B receptor was carried by astrocytes aggregating in neural lesions. In light of our findings, the possibility that astrocytes can be activated by the endothelin B receptor in response to neural tissue repair after damage to neurons would have to be considered.
    Neuroscience 12/1997; 81(2):565-77. DOI:10.1016/S0306-4522(97)00175-9 · 3.36 Impact Factor
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    ABSTRACT: 1. Our method of real-time monitoring of dopamine release from rat striatal slices revealed that endothelin (ET)-3-induced dopamine release was inhibited by N G-methyl-L-arginine (L-NMMA; 1 mM), an inhibitor of nitric oxide (NO) synthase, while N G-methyl-D-arginine (D-NMMA; 1 mM), an inactive isomer of L-NMMA, had no effect.2. The inhibition of L-NMMA (0.1 mM) became apparent when tissues were pretreated with tetrodotoxin (1 M) for 30 min and subsequently exposed to ET-3 (4 M).3. L-NMMA (0.1 and 1 mM) dose dependently protected against ET-3-triggered hypoxic/hypoglycemic impairment of striatal responses to high K+.4. Thus, NO may work as a promoter in mediation of the stimulatory and neurotoxic action of ET-3 on the striatal dopaminergic system, presumably by interacting with interneurons in the striatum.
    Cellular and Molecular Neurobiology 09/1997; 17(5):471-481. DOI:10.1023/A:1026354720732 · 2.51 Impact Factor
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    ABSTRACT: Endothelin (ET) receptors, ET-1-like immunoreactivity and nitric oxide synthase (NOS) were examined in the brain of stroke-prone spontaneously hypertensive rats (SHRSPs) with cerebral apoplexy. Our receptor autoradiographic method with 125I-ET-1 and unlabeled selective ligands for ET receptors revealed de novo expressions of ETA and ETB receptors in areas of neural lesions with cerebrovascular damage in SHRSPs. Immunohistochemical staining for ET-1 showed clear ET-1-like immunoreactivity in areas with highly expressed ET receptors. Histochemical studies on astrocytes and microglia suggested that these glial cells, aggregating in lesions, may carry ET receptors, ET-1-like immunoreactivity. Furthermore, NOS detected histochemically using an NADPH-diaphorase staining method was rich on glial cells in damaged areas of the brain in SHRSPs with cerebral apoplexy. Our data suggest the pathophysiological significance of glial ETA and ETB receptors, ET-1 and NOS in neural lesions of SHRSPs.
    Brain Research 06/1997; 756(1-2-756):61-67. DOI:10.1016/S0006-8993(97)00118-2 · 2.84 Impact Factor
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    ABSTRACT: 1. The receptor autoradiographic method done on the rat lower brain stem and cerebellum plus 125I-endothelin-1, BQ-123, an antagonist for the endothelin ETA receptor, and sarafotoxin S6c, an agonist for the ETB receptor, revealed minute amounts of the ETA receptor coexisting with the ETB receptor in the caudal solitary tract nucleus of the rat lower brain stem. 2. The ETB receptor is present predominantly in other parts of the lower brain stem. 3. Knowledge of the heterogeneous distribution of the central endothelin receptor subtypes aids in understanding the neurophysiology of endothelins.
    Cellular and Molecular Neurobiology 03/1997; 17(1):151-6. DOI:10.1023/A:1026341423825 · 2.51 Impact Factor
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    ABSTRACT: 1. We used the quantitative receptor autoradiographic method plus 125I-endothelin-1 (125I-ET-1), BQ-123, a specific antagonist for the endothelin ETA receptor, and sarafotoxin S6c, a selective agonist for the ETB receptor to investigate the ET receptor in the rat pituitary gland. 2. The method revealed that the BQ-123-sensitive ETA receptor was present predominantly in the anterior lobe and Rathke's pouch. 3. The posterior lobe contained BQ-123-sensitive ETA and sarafotoxin S6c-sensitive ETB receptors, in almost the same proportion. There was no significant 125I-ET-1 binding to the intermediate lobe. 4. Knowledge of the heterogeneous distribution of ET receptor subtypes in the pituitary gland supplies information that will be pertinent to physiological investigations of the gland.
    Cellular and Molecular Neurobiology 03/1997; 17(1):89-100. DOI:10.1023/A:1026381105170 · 2.51 Impact Factor

  • Japanese Heart Journal 01/1997; 38(4):597-597. DOI:10.1536/ihj.38.597 · 0.40 Impact Factor

  • Japanese Heart Journal 01/1997; 38(4):592-592. DOI:10.1536/ihj.38.592 · 0.40 Impact Factor

  • European Neuropsychopharmacology 06/1996; 6:129-129. DOI:10.1016/0924-977X(96)87900-4 · 4.37 Impact Factor
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    ABSTRACT: 1. When delayed neuronal death occurred in the hippocampus CA1 pyramidal cell layer of stroke-prone spontaneously hypertensive rats (SHRSP) at 4 and 7 days after a 10 min bilateral carotid occlusion and reperfusion, intense endothelin-1 (ET-1)- and ET-3-like immunoreactivities became evident in astrocytes in the damaged hippocampus CA1 subfields. 2. We also observed that microglia equipped with an ETb receptor aggregated within the CA1 pyramidal cell layer with neuronal death. 3. There was a dramatic increase in nitric oxide synthase (NOS) activity in astrocytes and microglia in the damaged hippocampus CA1 subfields. 4. Thus, the possibility that microglia with the ETb receptor are activated to produce NO, a neurotoxic factor, by astrocytic ET-1 and ET-3 produced in response to transient forebrain ischaemia would have to be considered.
    Clinical and experimental pharmacology & physiology. Supplement 01/1996; 22(1):S277-8. DOI:10.1111/j.1440-1681.1995.tb02915.x
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    ABSTRACT: 1. We characterized specific 125I-endothelin-1 (125I-ET-1) binding sites in microvessels isolated from human meningiomas, using an in vitro quantitative receptor autoradiographic technique coupled to a radioluminographic imaging plate system. 2. This newly developed and highly sensitive method revealed high-affinity ET receptors present in pellet sections of the microvessels from all the meningiomas studied, regardless of histological subtypes (dissociation constant, 1.2 +/- 0.3 nM; maximum binding capacity, 185 +/- 56 fmol/mg; means +/- SE for nine tumors). 3. In five cases of meningiomas, ET-3 competed for 125I-ET-1 binding to microvessels from those tumors with a low affinity [50% inhibiting concentration (IC50) of 1.6 +/- 0.4 x 10(-6) M], and a selective ETB receptor agonist, sarafotoxin S6c, up to 10(-6) M, did not displace ET binding from the sections. 4. In the sections of microvessels from four other tumors, biphasic competition curves were obtained in the case of incubation in the presence of increasing concentrations of ET-3, with an IC50 of 1.1 +/- 0.2 x 10(-9) M for the high-affinity component and 1.6 +/- 0.3 x 10(-6) M for the low-affinity component, respectively. In addition, S6c competed for ET binding to those sections (IC50 = 2.3 +/- 0.2 x 10(-10) M) and 10(-6) M S6c displaced 30% of the control, corresponding to the high-affinity component of competition curves obtained in the presence of ET-3. 5. Our results suggest that (a) capillaries in human meningiomas express a large number of high-affinity ETA (non-ETB) receptors with a small proportion of ETB receptors, and (b) ET may have a role in neovascularization, tumor blood flow, and/or function of the blood-tumor barrier in meningioma tissues by interacting with specific receptors present on the surface of the endothelium.
    Cellular and Molecular Neurobiology 07/1995; 15(3):327-40. DOI:10.1007/BF02089943 · 2.51 Impact Factor
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    ABSTRACT: We identified and characterized 125I-endothelin-1 (125I-ET-1) binding sites in tumor capillaries isolated from human glioblastomas, using the quantitative receptor autoradiographic technique with pellet sections. Quantification was done using the computerized radioluminographic imaging plate system. High-affinity ET receptors were localized in capillaries from glioblastomas and the surrounding brain tissues (KD = 4.7 +/- 1.0 x 10(-10) and 1.6 +/- 0.3 x 10(-10) M, respectively; Bmax = 161 +/- 38 and 140 +/- 37 fmol/mg, respectively; mean +/- SEM, n = 5). BQ-123, a selective antagonist for the ETA receptor, potently competed for 125I-ET-1 binding to sections of the microvessels with IC50 values of 5.1 +/- 0.3 and 5.1 +/- 1.5 nM, and 10(-6) M BQ-123 displaced 84 and 58% of ET binding to capillaries from tumors and brains, respectively. In addition, competition curves obtained in the presence of increasing concentrations of ET-3 showed two components (IC50 = 5.7 +/- 2.5 x 10(-10) and 1.4 +/- 0.2 x 10(-6) M for tumor microvessels, 1.8 +/- 0.6 x 10(-10) and 1.1 +/- 0.3 x 10(-6) M for brain microvessels, respectively). Our results indicate that (a) the method we used is simple and highly sensitive for detecting and characterizing various receptors in tumor capillaries, especially in the case of a sparse specimen, and (b) capillaries in glioblastomas express specific high-affinity ET binding sites, candidates for biologically active ET receptors, which predominantly belong to the ETA subtype.
    Journal of Neurochemistry 01/1995; 63(6):2240-7. DOI:10.1046/j.1471-4159.1994.63062240.x · 4.28 Impact Factor

Publication Stats

670 Citations
97.35 Total Impact Points


  • 1988-2002
    • Nagasaki University Hospital
      Nagasaki, Nagasaki, Japan
  • 1988-1998
    • National Institute of Mental Health (NIMH)
      • • Section on Pharmacology
      • • Laboratory of Clinical Science
      베서스다, Maryland, United States