Biomedical subjects

T Inagami

Publications and source records attributed to T Inagami.

At least 325 records · Page 18Linked to original sources

PubMed1986

Atrial natriuretic factor inhibits norepinephrine release evoked by sympathetic nerve stimulation in isolated perfused rat mesenteric arteries.

The effect of atrial natriuretic factor (ANF) on [3H]norepinephrine release evoked by sympathetic nerve stimulation was examined in the isolated perfused rat mesenteric arteries. ANF (1 nM to 0.1 microM) caused a dose-dependent inhibition of [3H]norepinephrine release during nerve stimulation. The present result indicates that ANF inhibits noradrenergic neurotransmission in the rat mesenteric arteries through a prejunctional mechanism. This prejunctional effect of ANF may in part contribute to its vasodilation action.

Animals↗

PubMed1986

Release of immunoreactive atrial natriuretic factor from rat hypothalamus in vitro.

Atrial natriuretic factor (ANF) had been found in brain tissues. Its role and the mechanisms by which it is produced and functions in the brain were not clear. We have initiated in vitro studies to find whether it is released from brain tissue and to elucidate the mechanism of its release. ANF was found to be released from rat hypothalamus by a depolarizing concentration of potassium and by a calcium-dependent mechanism. The ANF released was found to be predominantly a low molecular weight form. A small amount of high molecular weight form was also released. These results suggest that ANF produced in brain tissues is released, by a depolarization-induced and calcium-mediated mechanism, presumably from neuronal cells.

Animals↗

PubMed1986

Rat atrial natriuretic factor suppresses proopiomelanocortin-derived peptides secretion from both anterior and intermediate lobe cells and growth hormone release from anterior lobe cells of rat pituitary in vitro.

Synthetic rat atrial natriuretic factor (ANF) was found to attenuate, in a dose-dependent manner, basal and corticotropin-releasing factor-induced secretion of proopiomelanocortin-derived peptides from cultured anterior and intermediate lobe cells of rat pituitary. ANF was also found to suppress basal and growth hormone-releasing factor-stimulated secretion of growth hormone from anterior lobe cells of rat pituitary. These results, together with reports of the existence of ANF-positive neurons in the hypothalamus and ANF-positive fibers in the median eminence, suggest that hypothalamic ANF is probably involved in the regulation of pituitary hormone secretion, especially that of proopiomelanocortin-derived peptides and growth hormone.

Adrenocorticotropic Hormone↗

PubMed1986

Regulation of renin angiotensins by gonadotropic hormones in cultured murine Leydig tumor cells. Release of angiotensin but not renin.

Renin and angiotensins coexist in various tissues. The mode of control of the extrarenal renin-angiotensin system is not clear. Whether it is renin or angiotensin that is secreted has not been identified. We have investigated gonadotropin-dependent synthesis and subsequent release of the components of the intracellular renin-angiotensin system in a cloned and cultured mouse Leydig tumor cell line (MA-10). Treatment of cultured Leydig cells with bovine luteinizing hormone (bLH, 100 ng/ml) or human chorionic gonadotropin (hCG, 25 ng/ml) resulted in greater than 150- and 40- fold increased formation of angiotensin I and angiotensin II. In cells incubated with bLH or hCG, the majority of AII (up to 90%) was found in the culture medium while most of angiotensin I (greater than 85%) was in the cell lysate. Treatment with gonadotropic hormones (bLH/ hCG) increased renin 35- to 40-fold. Renin activity was confined mainly in the cell lysate even after the stimulation by gonadotropins, and only 1-2% of the total renin activity was detectable in culture medium. These results were interpreted that, in these transformed cells, hormonally-induced renin functions to generate angiotensin I within the Leydig cell and it is the angiotensins which are secreted.

Angiotensin I↗

PubMed1986

Generation of angiotensinogen by cultured neuroblastoma and glioma cells.

Cultured neuroblastoma cells and neuroblastoma-glioma cells have been shown to contain renin activity, angiotensin-converting enzyme activity, and angiotensins. It has been assumed that these cells also produce angiotensinogen as the substrate of an intracellular renin-angiotensin system. However, measurements of angiotensinogen have not been reported in the neuroblastoma or neuroblastoma-glioma cells, and the possibility that the cells generate angiotensins from fetal bovine angiotensinogen has not been eliminated. In this work angiotensinogen was shown to accumulate in the serum-free medium of thoroughly washed neuroblastoma cells (mouse Neuro-2A and rat B103) and glioma cells (rat C6). Separate experiments demonstrated that mouse Neuro-2A cells continue to produce angiotensinogen even after two passages in a defined serum-free culture medium. Further evidence that the angiotensinogen was not a contaminant from fetal bovine serum was obtained by the use of a monoclonal antibody raised against angiotensinogen of rat plasma. The angiotensinogen of Neuro-2A and C6-glioma cells is bound by the monoclonal antibody, whereas fetal bovine angiotensinogen is not bound. These results are consistent with the hypothesis that angiotensinogen is produced locally in the brain and in neuroblastoma cells as a substrate for an intracellular renin-angiotensin system.

Angiotensinogen↗

PubMed1986

Atrial pressure and secretion of atrial natriuretic factor into the human central circulation.

Atrial natriuretic factor, a peptide found in mammalian cardiac atria, has natriuretic and vasodilatory properties that may be important in the regulation of intravascular volume. To study factors related to its release in human subjects, intracardiac pressures and plasma atrial natriuretic factor concentrations in the central circulation were measured in 34 patients with a variety of cardiovascular disorders. Plasma atrial natriuretic factor concentration increased from the inferior vena cava to the right atrium (76 +/- 24 to 162 +/- 37 pg/ml, p less than 0.001) and from the vena cava to the aorta (76 +/- 24 to 177 +/- 46 pg/ml, p less than 0.001). Mean right atrial pressure was positively correlated with atrial natriuretic factor concentration in the pulmonary artery (r = 0.58, p less than 0.001), and mean pulmonary capillary wedge pressure was positively correlated with concentration in the aorta (r = 0.64, p less than 0.001). In six patients whose atrial natriuretic factor concentrations were measured at two different levels of atrial pressure, increased atrial pressure was accompanied by increased atrial natriuretic factor concentration in the pulmonary artery (p less than 0.01) and aorta (p less than 0.01). Atrial natriuretic factor levels measured in fresh myocardium from a patient undergoing cardiac transplantation showed tissue concentrations in the atria 500-fold higher than tissue concentrations in the ventricles. These data document that atrial natriuretic factor is found in human atrial myocardium and suggest that it may be released in response to increased atrial pressure. Such a secretory release mechanism is consistent with the hypothesis that atrial natriuretic factor plays a role in the regulation of circulatory volume.

Adult↗

PubMed1986

Immunocytochemical localization of renin and kallikrein in the rat renal cortex.

Immunocytochemical studies in the past, using alternate serial sections to localize individual antigens, concluded that there was no close relationship between renin- and kallikrein-containing structures in the rat kidney. We have investigated this relationship by simultaneously localizing renin and kallikrein in the same section using immunoperoxidase with two different chromogens. Analysis of serial kidney sections from three rats indicated that kallikrein-containing late distal tubular cells corresponded in their distribution to connecting tubule cells. They were observed in the proximity (less than 3 micrograms) of renin-containing JG cells in 66.6% of the superficial (N = 30), 46.6% of the midcortical (N = 15) and 26.7% of the juxtamedullary (N = 15) afferent arterioles surveyed. When traced through serial sections, 90% of the afferent arterioles from superficial glomeruli (N = 30), 86.7% of the afferent arterioles from midcortical glomeruli (N = 15) and 73.3% of those from juxtamedullary glomeruli (N = 15) came within 3 micrograms of a late distal tubule showing some kallikrein-positive cells. These cells were adjacent to the afferent arteriole in 67 to 80% of the arterioles surveyed. This spatial relationship suggests an anatomical basis for a possible interaction between the afferent arteriole, containing renin-positive JG cells, and kallikrein-positive late distal tubular cells.

Animals↗

PubMed1986

Increased concentration of plasma immunoreactive atrial natriuretic factor in Dahl salt sensitive rats with sodium chloride-induced hypertension.

In order to determine whether there is a relationship between genetically determined salt-induced hypertension and atrial natriuretic factor (ANF), a radio-immunoassay for ANF was applied to the determination of immunoreactive ANF in plasma, atrium, hypothalamus and pons of Dahl salt-sensitive (S) and -resistant (R) rats which were fed high- or low-salt diet for 7 weeks. A twofold higher concentration of plasma ANF was observed in high-salt S rats, which developed hypertension, compared with low-salt S rats or R rats on high or low salt, which were normotensive. No significant difference was seen in atrial concentrations of ANF between S and R rats. The brain ANF concentration of the high-salt group was lower than that of the low-salt group in both S and R rats. It is proposed that the elevation of plasma ANF in the hypertensive rats may reflect a compensatory mechanism induced by volume expansion in the salt-fed S rats.

Animals↗

PubMed1986

Role of vascular angiotensin II released by beta-adrenergic stimulation in rats.

The effect of a beta-adrenoceptor agonist on the release of the components of the vascular renin-angiotensin system was examined in vitro. Isolated rat mesenteric arteries were perfused in an open system with Krebs-Ringer solution and released immunoreactive angiotensin II (ANG IIir) into the perfusate was directly determined using a Sep-Pak C-18 cartridge connected to the perfusion system. Renin activity in the concentrated perfusate was also determined. Isoproterenol (1 nM-1 microM) increased the release of ANG IIir in a dose-dependent manner. The increase in ANG IIir release during isoproterenol (1 microM) infusion was inhibited by propranolol (1 microM) or captopril (2 microM). Isoproterenol-induced increment of ANG IIir release was blocked by the selective beta 2-adrenoceptor antagonist, ICI 118,551 (1 microM), but not by the selective beta 1-adrenoceptor antagonist, atenolol (1 microM). Renin activity in the perfusate was measurable, but did not increase in response to isoproterenol (1 microM) infusion. There was no significant difference in the response of ANG IIir release to isoproterenol between spontaneously hypertensive rats and Wistar-Kyoto rats. The present results indicate that locally generated ANG II is released by beta 2-adrenoceptor activation. The beta-adrenoceptor agonist and the vascular renin-angiotensin system may play an important role for the regulation of peripheral vascular tone.

Adrenergic beta-Antagonists↗

PubMed1986

Subcellular localization of angiotensin-converting enzyme in cultured neuroblastoma cells.

Angiotensin-converting enzyme (ACE) activity of Neuro-2A mouse neuroblastoma cells was found predominantly in particulate fractions. Density gradient centrifugation of the particulate fractions showed ACE activity in light fractions of the gradient, a result suggesting a plasma membrane localization. This was confirmed using the aqueous two-phase polymer system of plasma membrane isolation. The rapid and energy-independent hydrolysis of exogenous substrate by ACE of intact cells and the sensitivity of the enzyme of intact cells to proteases indicate further that the active site of ACE is oriented extracellularly.

Animals↗

PubMed1986

Beta-adrenoceptor-mediated release of angiotensin II from mesenteric arteries.

Essential components of the renin-angiotensin system such as renin enzymes, angiotensinogen, converting enzyme, and angiotensin receptors have been found in vascular tissues. Locally generated angiotensin (ANG) II may regulate vascular tone by contracting vascular smooth muscle or potentiating sympathetic activity. Recently it was suggested that beta-adrenoceptor-induced enhancement of noradrenergic neurotransmission is mediated by the vascular renin-angiotensin system. The present study was designated to obtain direct evidence for the release of ANG II from the vasculature by beta-adrenoceptor activation. Isolated rat mesenteric arteries were perfused in vitro with Krebs-Ringer solution, and released ANG II was concentrated in a Sep-Pak C-18 cartridge connected to the perfusion system. High-pressure liquid chromatography combined with radioimmunoassay clearly demonstrated the presence of ANG I, II, and a small amount of ANG III in the perfusate. Isoproterenol (10(-9) - 10(-6) M) induced the enhancement of pressor responses to nerve stimulation. This effect was markedly suppressed by propranolol (5 X 10(-7) M), captopril (2 X 10(-6) M), or [Sar1-Ile8]ANG II (10(-6) M). Isoproterenol (10(-9) - 10(-6) M) caused increase in the release of ANG II from mesenteric arteries. The increase in ANG II release during isoproterenol (10(-6) M) infusion was blocked by propranolol (10(-6) M). Captopril (2 X 10(-6) M) also inhibited the increase in ANG II induced by isoproterenol. These results indicate that locally generated ANG II is released from isolated perfused rat mesenteric arteries and its release is mediated by beta-adrenoceptors.

Angiotensin II↗

PubMed1986

Sodium loading and posture modulate human atrial natriuretic factor plasma levels.

Atrial natriuretic factor is postulated to act through atrial stretch receptors as a volume regulatory hormone that stimulates diuresis and natriuresis in response to increased atrial pressure. To characterize the stimuli associated with the release of atrial natriuretic factor in humans, we studied 14 normal subjects, both in the supine position and after 10 minutes in an upright posture, while they were on a regular diet (Day 0) and during 3 days of supplemental sodium chloride intake (8 g/day). Radioimmunoassay of plasma atrial natriuretic factor was performed with rabbit antibody to the human hormone amino acids (102-126). Urinary sodium excretion increased from 111 +/- 13 mEq/day (mean +/- SEM) on Day 0 to 275 +/- 15 mEq/day by the third day (Day 3) of high sodium intake. The level of atrial natriuretic factor in the supine position rose from 17 +/- 4 pg/ml (Day 0) to 76 +/- 13 pg/ml on Day 3 (p less than 0.001) and after 10 minutes in an upright posture on Day 3, the level fell to 32 +/- 10 (p less than 0.005). Plasma concentrations of atrial natriuretic factor correlated positively with spot and 24-hour urinary sodium excretion and weight gain, and correlated negatively with plasma aldosterone and renin activity. We conclude that the response of atrial natriuretic factor to sodium loading and posture change in humans is appropriate for a volume regulatory hormone.

Adult↗

PubMed1986

Vascular renin-angiotensin system in two-kidney, one clip hypertensive rats.

The possible role of the renin-angiotensin system in the maintenance of hypertension in two-kidney, one clip hypertensive rats was studied. Plasma renin activity rose rapidly and markedly in association with the elevation of blood pressure and then decreased gradually, although blood pressure remained high. Renin activity in the lung, aorta, and mesenteric artery also increased with the development of hypertension and then decreased in a way similar to that of plasma renin activity at the chronic stage of hypertension. Plasma angiotensin converting enzyme activity did not change significantly until 16 weeks after unilateral renal artery clipping, whereas vascular angiotensin converting enzyme activity significantly increased at the chronic, but not the acute, stage of hypertension. In chronically renal hypertensive rats, 1-sarcosine, 8-isoleucine angiotensin II or enalapril, an angiotensin converting enzyme inhibitor, lowered the blood pressure and enalapril also lowered the angiotensin converting enzyme activity of vascular tissues. The constrictor effect of angiotensin I was greater in isolated arteries from chronically hypertensive rats than in those from age-matched normotensive rats. These results suggest that the vascular renin-angiotensin system plays an important role in the maintenance of two-kidney, one clip hypertension. Elevated vascular angiotensin converting enzyme activity appears to increase local production of angiotensin II, which results in vasoconstriction by acting directly and indirectly through adrenergic nerves on vascular smooth muscle.

Angiotensin II↗

PubMed1986

The storage form of renin in renin granules from rat kidney cortex.

Renin granules were partially purified from rat kidney cortex, and a storage form of renin in the granules was examined. Renin granules were isolated by discontinuous Percoll density gradient centrifugation followed by continuous Percoll density gradient centrifugation. The partially purified fraction was free from mitochondria and microsomes, as judged by the absence of marker enzymes of these organelles, but contained some lysosomal enzyme activities. The specific renin activity was 0.58 mg angiotensin I/hr/mg protein, 500 times as active as the original homogenate. Immunochemical staining with specific antisera against rat kidney renin revealed that about 10% of the granules recovered in the partially purified fractions were stained strongly. The stored renin was not activated either by acidification or by trypsin treatment, indicating that stored renin was in the fully active form. By sodium dodecyl sulfate gel electrophoresis, the stored renin had two different molecular weights, 38,000 and 36,000, and these molecular weights were not reduced by dithiothreitol or 2-mercaptoethanol, suggesting that these renins are single-chain types as opposed to the two-chain type found in male mouse submaxillary gland. These results suggest that active renins with two different molecular weights may be released from renin granules of juxtaglomerular cells.

Animals↗

PubMed1986

Renin in the rat pituitary coexists with angiotensin II and depends on testosterone.

In the rat pituitary gland, immunoreactive angiotensin II (ANG II), renin, and LH, but not PRL, were found within the same cells of the anterior pituitary gland by staining with the avidin-biotin complex method in adjacent sections. No renin-positive staining was observed in the pituitary of the rats after 10 days of castration, but positive staining reappeared after 8 weeks. This effect of castration on renin immunoreactivity was abolished by the simultaneous administration of testosterone. In contrast, ANG II immunoreactivity was unaffected by castration. The intensity of renin immunoreactivity in the pituitary was less prominent in the female than in the male rat. These results suggest that there exists a pituitary renin-angiotensin system localized in the gonadotrophs and that the pituitary renin is under androgenic control.

Angiotensin II↗

PubMed1986

Is renin secreted by exocytotic mechanism through mature renin granules from juxtaglomerular cells?

Mature renin granules were isolated by the combination of discontinuous and continuous Percoll density gradient centrifugation. Stored renin in the renin granules was found to consist of isoelectrically seven different forms. The seven different isoelectric points (pIs) were 5.6, 5.35, 5.2, 5.0, 4.8, 4.6 and 4.4. Approximately 70% of the stored renin as the total enzymatic activities from all isoelectric peaks was found in a peak which pI corresponded to be 5.35. Renin secreted from isolated glomeruli was also focused into seven peaks possessing identical values. However, the distribution pattern of renin peaks was quite different from that of stored renin. In the secreted renin, peaks of 5.35 (pI) and 5.2 (pI) showed high renin activity and each had approximately 30% of released renin as the total recovered. These results indicate multiple forms of renin are stored and secreted by rat kidney. As the distribution pattern of enzymatic activities in renin peaks between stored renin and secreted renin are different, it is probable that renin may not secreted through mature renin granules by exocytotic mechanism.

Animals↗

PubMed1986

Alterations in atrial and plasma atrial natriuretic factor (ANF) content during development of hypoxia-induced pulmonary hypertension in the rat.

Distension of the atrial wall has been proposed as a signal for the increased release of atrial natriuretic factor (ANF) from atrial myocytes in response to perceived volume overload. To determine whether pressure changes resulting from hypertension in the pulmonary circulation may stimulate release of ANF, rats were exposed to chronic hypobaric hypoxia for 3 or 21 days and the ANF concentration in the atria and plasma were determined by specific radioimmunoassay. Exposure to chronic hypoxia resulted in significant increases in hematocrit at both 3 (p less than 0.025) and 21 days (p less than 0.005) and in the development of right ventricular hypertrophy (RVH) expressed as the ratio of the weight of the right ventricle to the weight of the left ventricle and septum (RV/LV+S) at both 3 (RV/LV+S = 0.278 +/- 0.005) and 21 days (RV/LV+S = 0.536 +/- 0.021). After 21 days, left atrial (LA) ANF content was significantly increased in hypoxic rats compared to controls (508 +/- 70 ng/mg tissue vs 302 +/- 37 ng/mg), while right atrial (RA) ANF content was significantly reduced (440 +/- 45 vs 601 +/- 58 ng/mg). At this time, plasma ANF concentration was significantly elevated compared to controls (238 +/- 107 pg/ml vs 101 +/- 10 pg/ml). These results suggest that the development of pulmonary hypertension following chronic hypobaric exposure induces altered atrial ANF content and increased plasma ANF concentration as a result of altered distension of the atrial wall.

Animals↗

PubMed1986

Renin in angiolymphoid hyperplasia with eosinophilia. Its possible effect on vascular proliferation.

A hypertensive man became normotensive after the surgical removal of two subcutaneous masses of angiolymphoid hyperplasia with eosinophilia. To demonstrate that angiolymphoid hyperplasia with eosinophilia is a renin-producing pathologic condition, Bowie stain for juxtaglomerular cell granules and immunohistochemistry for human renin were used. Bowie stain was positive in cells showing cytoplasmic granules similar to those found in the juxtaglomerular cells of the kidney. Immunohistochemical staining using antiserum against human renin showed the presence of renin-containing cells. This staining was not seen after substitution of the specific renin antiserum by preimmune serum, by the renin antiserum preabsorbed with pure human renin, or with plasma from a patient with high-plasma renin activity. Renin-containing cells were located in areas surrounding vascular structures and were apparently neither endothelial, mast, nor lymphoid cells. Six of eight additional cases of angiolymphoid hyperplasia with eosinophilia were positive for renin-containing cells. Renin has been described in several other histologically highly vascularized tumors. Since a product of renin, angiotensin II, has been found to have angiogenic properties, it is possible to postulate that renin, through angiotensin II, may stimulate the proliferation of vessels and, therefore, may be involved in the pathogenesis of angiolymphoid hyperplasia with eosinophilia.

Abdominal Muscles↗