Biomedical subjects

A Ohtsuka

Publications and source records attributed to A Ohtsuka.

At least 91 records · Page 5Linked to original sources

PubMed1995

Blood vascular bed and pericapillary space in rat parathyroid glands.

The blood vascular bed and pericapillary space of the rat parathyroid gland were studied by scanning electron microscopy of vascular casts, freeze-cracked tissue blocks, and NaOH-treated tissue specimens. The findings were supplemented by transmission light and electron microscopy of sectioned tissue samples. The rat parathyroid gland contained a rich network of freely anastomosing capillaries. These capillaries were surrounded by marked pericapillary spaces that were demarcated by basal lamina of both capillaries and parenchymal cells. The pericapillary spaces contained many collagen fibrils and frequently issued some projections running deep into the sheets of parathyroid cells. The latter projections may be useful to supply the parenchymal cells located far from the capillaries. The collagen fibrils may regulate the flow of tissue fluid in the pericapillary space and convey parathyroid hormone, which is released at the apicolateral domain, into the capillaries.

Animals↗

PubMed1995

Neurons with perineuronal sulfated proteoglycans in the human visual cortex, with special reference to their reactions to lectins.

The human visual cortex, especially its ganglionic lamina, was found to contain many neurons with perineuronal sulfated proteoglycans which were stained with cationic iron colloid and aldehyde fuchsin. It also contained many neurons with surface glycoproteins labeled with lectin Vicia villosa agglutinin (VVA) or Glycine max agglutinin (SBA). Double staining frequently showed that the neurons stained with cationic iron colloid were not labeled with lectin VVA or SBA. Hyaluronidase and chondroitinase ABC/heparitinase/keratanase digestions eliminated the perineuronal cationic iron colloid reaction, but never interfered with the cell surface lectin labeling. These findings indicate that the cell surface glycoproteins reactive to lectin VVA or SBA are neither structural elements nor adhesive molecules of the proteoglycans. Double staining further demonstrated that in some neurons with perineuronal sulfated proteoglycans, the cytoplasm was labeled with lectin Arachis hypogaea agglutinin (PNA). It was further noticed that the lectin VVA-labeled neurons were not always identical with the neurons labeled with lectin SBA or with lectin PNA.

Colloids↗

PubMed1995

Strongly anionic sites in peripheral axons of the rat sciatic nerve: light and electron microscopic detection using cationic colloidal iron.

Anionic sites in the rat sciatic nerve were studied by light and electron microscopy using a fine-granular cationic colloidal iron staining method (Murakami et al., 1986). The axon, as well as the endoneurium, the epineurium and the basement membrane of Schwann cells, were all confirmed to react strongly to the cationic colloidal iron even at a pH value of 1.0-2.0. Prior hyaluronidase digestion decreased the colloidal strain of the epineurium; chondroitinase ABC weakened that of the endoneurium and the basement membrane of Schwann cells. However, as axons retained stainability with cationic colloidal iron even after combined digestion with hyaluronidase, chondroitinase ABC, heparitinase and keratanase, the authors consider sulfated glycoconjugates and not those substances which are digestible with such common enzymes. The acid groups ionized at pH 1.0 are most likely sulfate groups. Methylation deprived the axon of the reactivity to cationic colloidal iron staining, and even subsequent saponification could not recover this reactivity to its full extent. In the axon, electron microscopy revealed a deposition of colloidal iron on the external surfaces of microtubules and neurofilaments in the axoplasm and of very fine filaments connecting them. This highly negatively charged intra-axonal network could also serve toward a supportive function in maintaining the spatial distribution of microtubules either mechanically or through electrostatic repulsion or, possibly, serve as an intra-axona cation exchange reservoir.

Animals↗

PubMed1995

Perineuronal sulfated proteoglycans and dark neurons in the brain and spinal cord: a histochemical and electron microscopic study of newborn and adult mice.

Neurons of intracerebellar nuclei in the mouse brain were demonstrated to possess a marked surface coat, formed 3-4 weeks after birth, which was stainable with cationic iron colloid or aldehyde fuchsin. Neurons with a similar surface coat were noted as relay or local interneurons in rather restricted areas such as the occipital cortex, retrosplenial cortex, zona incerta, hippocampal subiculum and spinal posterior horn. Dark neurons with condensed cytoplasm were also shown to be covered with the surface coat. The surface coat was stained doubly with cationic iron colloid and aldehyde fuchsin. Digestion with hyaluronidase eliminated the stainability of the surface coat to both agents. Combined digestion with chondroitinase ABC, heparitinase and keratanase eliminated the cationic iron colloid staining of the surface coat, but did not interfere with the aldehyde fuchsin staining of the surface coat. Electron microscopy of ultrathin sections revealed that the iron particles indicating sulfated proteoglycans were preferentially deposited in the perineuronal tissue spaces. Many neurons in the hippocampal subiculum possessed cell surface glycoproteins which were labeled with lectin Vicia villosa or soybean agglutinin and formed 1-2 weeks after birth. Double staining revealed that these lectin-labeled neurons were identical in part with the neurons reactive to the cationic iron colloid. Dark neurons began to appear 3-4 weeks after birth. The formation of perineuronal sulfated proteoglycans and the appearance of dark neurons, both occurring during the weaning period, may reflect the morphological and physiological completion of the brain. Dark neurons are suggested to be exhausted cells that are restored to light or normal neurons after sleep.

Aging↗

PubMed1995

Blood microvascular organization of the nasal-associated lymphoid tissue of the guinea pig: a scanning electron microscopic study of corrosion casts.

It has previously been confirmed that the guinea pig has aggregations of 10-20 lymphoid follicles at the junction of the nasal cavity and the nasopharyngeal duct. The vascular architecture of this nasal-associated lymphoid tissue (NALT) was studied by the corrosion cast/scanning electron microscope method. The NALT was supplied by branches of the inferior nasal artery. These afferent arterial branches gave off arterioles to the follicles and the interfollicular regions, where the arterioles ramified into capillaries. Some of these arterioles reached the subepithelial region to form a single-layer dense capillary network. The subepithelial capillaries gathered into short collecting venules, which in turn drained into high endothelial venules (HEV) in the interfollicular region. The HEV, which also receives tributaries from the follicular and interfollicular capillary plexuses, descended in the interfollicular regions and finally flowed into the efferent veins at the bottom of the NALT. Indentations impressed by high endothelial cells (HEC) were prominent on the surface of the HEV casts, and their frequency was larger in the upper course or segments than in the lower. This suggests that the incidence of HEC in the upper segments is higher than in the lower segments, and these findings are consistent with the hypothesis that some substances which are taken up into the subepithelial capillaries and transported to the venules induce differentiation and maintain of HEVs.

Animals↗

PubMed1995

Calcitonin reduced corticosterone-induced muscle proteolysis.

Two-experiments were conducted to clarify the effect of calcitonin (CT) on growth, muscle protein breakdown, plasma corticosterone (CTC) concentration and urinary calcium excretion in young growing rats treated with CTC. Rats received hormonal treatments for 6 days (Experiment 1) or 24 h (Experiment 2). Dose levels of CTC were 10 mg and 5 mg/100 g body weight/day in experiment 1 and 2, respectively, and that of CT was 100 m unit/100 g body weight/day in both experiments. Muscle protein degradation was evaluated by urinary N tau-methylhistidine excretion. CT increased 24-h urinary calcium excretion but not 6-day calcium excretion. CTC markedly inhibited growth, accelerated muscle protein degradation and increased calcium excretion in both experiments. However, very interestingly, CT minimized CTC-induced muscle proteolysis, and normalized the CTC-induced decrease in urinary calcium excretion. Furthermore, CT decreased the CTC-induced increase in the plasma CTC concentration. The present observations indicate that CT reduces CTC-induced muscle protein breakdown by reducing the plasma CTC concentration and that increased urinary calcium excretion due to CT treatment may also play a role in reducing CTC-induced muscle protein breakdown.

Animals↗

PubMed1995

[Effects of a beta 2-agonist, sodium cromoglycate, and an anticholinergic agent on hyperventilation-induced bronchoconstriction in sensitized rabbits].

Hyperventilation can induce bronchoconstriction in ovalbumin-sensitized rabbits. To investigate the roles of the beta-receptor parasympathetic nervous system and of chemical mediators in hyperventilation-induced bronchoconstriction (HIB), the effects of a beta 2-agonist, of sodium cromoglycate, and of an anticholinergic agent on HIB were studied. Rabbits were divided to four groups and treated as follows. Group 1: Control (n = 7, 0.9% saline); Group 2: Procaterol (n = 4, 50 micrograms/l); Group 3: Sodium cromoglycate (n = 4, 10 mg/ml); and Group 4: Ipratropium bromide (n = 6, 1 mg/ml). Each drug was inhaled for 1 min via an ultrasonic nebulizer. Then, for the eucapnic hyperventilation challenge, sensitized rabbits were mechanically hyperventilated for 15 min (120 breaths/min, tidal volume = 7 ml/kg) with dry air containing 5% CO2 at room temperature. Total lung resistance and dynamic compliance were measured before, and 0, 5, 15, and 30 min after hyperventilation. The mean percent change in resistance measured 5 min after the hyperventilation was +49% in group 1, -6% in group 2, +23% in group 3, and +1% in group 4. The changes in groups 2 and 4 were significantly less than in group 1 (p < 0.05). In conclusion, HIB is mainly caused by bronchial smooth muscle constriction, and chemical mediators and the parasympathetic nervous system may play important roles in the development of HIB in sensitized rabbits.

Acetylcholine↗

PubMed1994

Effects of dietary corticosterone and trilostane on growth and skeletal muscle protein turnover in broiler cockerels.

1. The effects of dietary corticosterone and trilostane, an inhibitor of glucocorticoid synthesis, on: growth, rates of synthesis and breakdown of skeletal muscle protein, and content of abdominal fat were studied in broiler chickens. 2. Dietary corticosterone (5, 10 or 20 mg/kg) depressed body weight gain and increased abdominal fat content in a dose-dependent manner while dietary trilostane (1.4 or 7.0 mg/kg) had no effect. 3. The rate of protein breakdown in skeletal muscle estimated from N tau-methylhistidine excretion was increased in a dose-dependent manner by dietary corticosterone but it was decreased by trilostane. 4. The rate of skeletal muscle protein synthesis was not affected by corticosterone although it was decreased by trilostane. 5. Plasma corticosterone concentration was increased in a dose-dependent manner by dietary corticosterone and decreased by treatment with 7 mg trilostane/kg diet. 6. The results indicate that higher concentrations of plasma corticosterone increase protein breakdown in skeletal muscle but do not affect muscle protein synthesis while both the rates of synthesis and breakdown are decreased when plasma corticosterone concentration is reduced.

3-Hydroxysteroid Dehydrogenases↗

PubMed1994

Refractoriness of eucapnic hyperventilation-induced bronchoconstriction in rabbits.

The mechanism of refractoriness in bronchoconstriction after repeated hyperventilation was investigated in 18 sensitized rabbits. Rabbits were separated into three groups: an untreated control group (n = 7), a cimetidine-treated group (n = 6), and an indomethacin-treated group (n = 5). After anesthetization, hyperventilation was performed for 15 min (120 breaths/min, 7 ml/kg tidal volume) with dry air containing 5% CO2. Total lung resistance (RL) and dynamic compliance (Cdyn) were measured before (baseline) and after hyperventilation challenge. After RL and Cdyn had returned to baseline values, the hyperventilation challenge was repeated. In the control group maximal increase in percent RL (max %RL) was 49 +/- 9% after the first challenge, but 16 +/- 4% after the second challenge, indicating refractoriness. A similar tendency was observed in percent Cdyn. In the cimetidine- and indomethacin-treated groups, max %RL were 42 +/- 3% and 60 +/- 15% after the first challenge, and 35 +/- 8% and 60 +/- 7% after the second challenge, respectively, indicating no refractoriness. These results suggest that the H2-receptor and bronchodilating prostanoids play an important role in producing the refractoriness to bronchoconstriction observed in sensitized rabbits after repeated hyperventilation.

Airway Resistance↗

PubMed1994

Anionic sites on the free surface of the peritoneal mesothelium: light and electron microscopic detection using cationic colloidal iron.

To examine charged sites on the peritoneal free surface, cationic and anionic colloidal iron methods were applied. Light microscopy of the human and mouse specimens using cationic colloidal iron staining at pH 1.5-7.3 and successive ferrocyanide treatment resulted in a distinct Prussian blue reaction on the free surface of the parietal, mesenteric and visceral peritonea at all the examined pH values. In transmission electron microscopy of specimens stained with cationic colloidal iron at pH 1.5, colloidal particles accumulated in a dotted fashion on the free surface of the mesothelial cells. At pH 7.3, colloidal particles were deposited as numerous fine strands (100-300 nm in length), whose ends often attached to the luminal aspect of the mesothelial cell membrane. Anionic colloidal iron stain at pH 7.2 gave no deposition of colloidal particles on the mesothelial free surface. Priorly methylated samples lost stainability of the peritoneal free surface to cationic colloidal iron staining at pH 1.5 or 2.5, while methylated-saponified sections recovered it. Pretreatment with neuraminidase inhibited the cationic colloidal iron staining at pH 1.5 on the mesothelial free surface. These results indicate that the mesothelial surface anionic sites ionizing at pH 1.5 are mostly due to the carboxyl group of sialic acid. It is suggested that the peritoneal free surface substance stained with the cationic colloidal iron may be membrane-associated sialomucin, whose rich negative-charged sites may repulse each other to prevent peritoneal adhesion or to maintain peritoneal cavity.

Adolescent↗

PubMed1994

Neurons with intensely negatively charged extracellular matrix in the human visual cortex.

Neurons in the human visual cortex were demonstrated to possess an intensely negatively charged surface coat which was stained with cationic iron colloid and aldehyde fuchsin. Digestion with hyaluronidase eliminated both the iron colloid and fuchsin stainings of the coats. Treatment with chondroitinase ABC, heparitinase and keratanase eliminated the iron colloid staining of the coats, but did not interfere with the fuchsin staining. Electron microscopy of ultrathin sections revealed that the cationic iron particles were preferentially deposited in the perineuronal tissue spaces. These findings indicate that the surface coats consist of sulfated proteoglycans, which, as an extracellular matrix, occupy the perineuronal tissue spaces. This study further demonstrates that neurons with such surface coats are identical with neurons labeled with lectin Vicia villosa agglutinin. The cell surface glycoproteins reactive to this lectin may not be the structural elements of the sulfated coats since the lectin labeling was not interrupted by the hyaluronidase digestion.

Anions↗

PubMed1994

Microcirculatory patterns in human pancreas: supplementary observations of vascular casts by scanning electron microscopy.

The blood vascular bed of two human pancreata was replicated partially by arterial perfusion of intentionally reduced amounts of low viscosity methacrylate resin, to be observed with a scanning electron microscope. The findings were compared with those obtained from a pancreas replicated completely by a sufficient amount of resin. Complete replication confirmed our previous findings (MURAKAMI et al., 1992) that many exocrine lobules contained one or more endocrine islets, which preferentially issued insulo-acinar portal vessels continuous with the lobular capillaries. Incomplete replication demonstrated that the casting medium filled blood capillaries in the endocrine islets more promptly than those in the exocrine lobules and secretory ducts. Furthermore, islet-containing lobules allowed a more rapid resin flow to the exocrine tissue via the insulo-acinar portal route than did the lobules lacking an islet. Since the resin medium used had the viscosity of blood and was injected under physiological pressure, the results obtained by the incomplete arterial injections are believed to suggest the physiological state of blood flow in the pancreas.

Adult↗

PubMed1994

The occurrence of neurons with strongly negatively charged surface coats in mammalian, avian, reptilian, amphibian and piscine brains.

Neurons with strongly negatively charged surface coats were recognized in mammalian, avian, reptilian, amphibian and piscine brains. Many large-sized neurons had strongly negatively charged surface coats in the visual cortex and brain stem of the cow, cat, guinea pig, mouse, quail and parakeet. Such neurons were also seen in the brain stem of the lower vertebrates such as the house lizard, Japanese terrapin, bullfrog, newt, carp and sweetfish.

Amphibians↗

PubMed1994

[The role of parasympathetic nerve on hyperventilation-induced bronchoconstriction in sensitized rabbits].

We have demonstrated that hyperventilation-induced bronchoconstriction (HIB) is observed in sensitized rabbits, and that histamine may play a role in HIB. To investigate the role of parasympathetic neurotransmission in HIB, the effects of atropine and vagotomy on HIB were examined. Japanese white rabbits were sensitized with ovalbumin. They were then anesthetized and paralyzed with pancuronium, and then mechanically ventilated (TV: 7 ml/kg, RR: 40/min). The rabbits were divided into the following three groups: I) control group (n = 7), II) atropine-treated group (n = 6), III) bilateral vagotomy group (n = 6). Hyperventilation was performed at TV: 7 ml/kg, RR: 120/min for 15 min in dry room air containing 5% CO2. Total lung resistance (RL) and dynamic compliance (Cdyn) were measured before and at 0, 5, 15 and 30 minutes after hyperventilation. RL increases significantly at 5 minutes in group 1, but this change was significantly less marked in groups 2 and 3. However Cdyn was not significant change in each group. We conclude that parasympathetic neurotransmission may play an important role in HIB of sensitized rabbits.

Animals↗

PubMed1993

The effect of flow on the expression of vascular adhesion molecule-1 by cultured mouse endothelial cells.

Adherence of leukocytes to vascular endothelial cells (ECs) is known to be sensitive both to blood flow and adhesive proteins on EC surface. To elucidate the effect of blood flow on the surface expression of adhesive proteins, cultured ECs derived from mouse lymph nodes were exposed to different levels of wall shear stress in a flow-loading chamber, and changes in the expression of vascular adhesion molecule-1 (VCAM-1) and CD44 were evaluated by immunostaining with monoclonal antibodies and flow cytometry. Both proteins were expressed on non-activated cultured ECs. When exposed to flow with shear stress of 1.5 dynes/cm2 for 24 hr, VCAM-1 nearly disappeared on fluorescence micrographs, while CD44 showed no change. Flow cytometric analysis showed that the mean channel fluorescence of VCAM-1 was decreased about 75% by application of flow for 24 hr (p < 0.001), but that of CD44 remained unchanged. VCAM-1 expression began to decrease around 1 hr after the initiation of flow and became markedly reduced with time, reaching a minimum after 24 hr. When the cells subjected to flow for 24 hr were returned to stationary state, the reduced VCAM-1 expression was almost completely restored in 72 hr, indicating that the change was reversible. The magnitude of the reduction of VCAM-1 expression was also dependent on the intensity of the wall shear stress applied, ranging from 0 to 7.2 dynes/cm2. These results, demonstrating an explicit down-regulating effect of flow on VCAM-1 expression of cultured ECs, suggested preferential adhesion of leukocytes to ECs at low shear regions at the vascular wall.

Animals↗

PubMed1993

Wall shear stress rather than shear rate regulates cytoplasmic Ca++ responses to flow in vascular endothelial cells.

Recent evidence suggests that the vascular endothelial cell (EC) can sense the flow-rate over its surface and according to the information, regulates not only its own morphology and functions but also those of the surrounding smooth muscle and other tissues. There is now a discussion over which of the following mechanisms actually initiates the signal-transacting response of EC against flow: the mechanical shear deformation of the cell due to flow-oriented wall shear stress (tau), or the diffusional accumulation of vasoactive agonists on the cell surface modulated by wall shear rate (gamma) or both. To identify the relative importance of each mechanism, we examined quantitative changes in the cytoplasmic free Ca++ concentration ([Ca++]i) in cultured EC in the presence of the Ca++ mobilizing agonist ATP, i.e., a second messenger response of the internal signalling system, following the perfusion of two buffers with different viscosities (mu), which relates these factors as tau = mu gamma. The results of in vitro fluorescence photometry in EC with Fura-2 showed that the [Ca++]i level was enhanced with increase in the shear rate but to a greater extent with higher viscosity, and that the [Ca++]i levels at the same calculated level of shear stress were virtually identical, regardless of difference in shear rate and viscosity. This quantitative one-to-one relationship between the shear stress and the second messenger response suggests that wall shear stress rather than wall shear rate is the principal physical factor eliciting EC responses to flow.

Animals↗

PubMed1993

Close correlation between cytoplasmic Ca++ levels and release of an endothelium-derived relaxing factor from cultured endothelial cells.

We studied whether there is a quantitative relationship between free cytosolic Ca++ levels and the release of an endothelium-derived relaxing factor (EDRF) from cultured fetal bovine aortic endothelial cells (EC). EC pretreated with indomethacin were stimulated by the agonists adenosine triphosphate (ATP), bradykinin (BKN), acetylcholine (ACh) and calcium ionophore (A23187) in various concentrations (10(-8)-10(-5) M), and the amount of EDRF released was determined on the basis of endothelium-free rabbit aortic ring relaxation and cultured smooth muscle cell cGMP content. Changes in intracellular Ca++ concentration ([Ca++]i) in response to the same stimuli were determined by photometric fluorescence microscopy using the fluorescent calcium indicator Fura-2. EC stimulation by ATP and A23187 induced dose-dependent increases in both [Ca++]i and the amount of EDRF released. BKN increased both [Ca++]i and EDRF release upon initial exposure (10(-8)M), but there were no further changes at higher concentrations. ACh induced no significant changes in either [Ca++]i or EDRF release. There was a close quantitative correlation between agonist-induced changes in [Ca++]i and the amount of EDRF released (relaxation response in aortic rings and cGMP levels.) (p < 0.001) Removal of extracellular Ca++ eliminated continuous elevation in both [Ca++]i and the amount of EDRF induced by ATP (10(-5)M), BKN (10(-8)M) and A23187 (10(-6)M). These findings suggest that intracellular Ca++ levels are directly linked to the amount of EDRF released, and that extracellular Ca++ is essential for EDRF release because its influx is involved in the continuous elevation of [Ca++]i.

Acetylcholine↗

PubMed1993

The insulo-acinar portal and insulo-venous drainage systems in the pancreas of the mouse, dog, monkey and certain other animals: a scanning electron microscopic study of corrosion casts.

Scanning electron microscopy of vascular casts of the pancreas from monkies, cattle, pigs, dogs, cats, rabbits, guinea pigs and mice showed certain species differences in the occurrence of intralobular and interlobular islets and in the microcirculatory pattern of these islets. Interlobularly located islets were frequently found in the mouse and guinea pig, as has been previously established in the rat (Murakami and Fujita, 1992); they emitted insulo-venous efferent vessels directly draining into veins. In contrast, the intralobular islets in the guinea pig usually issued insulo-acinar portal vessels continuous with the lobular capillary network. In the mouse, they usually emitted both the insulo-acinar portal and insulo-venous efferent vessels. The insulo-venous efferent vessels, including those of the interlobular islets, could partly be portal in nature since they occasionally issued portal branches directed to the lobular capillary network. In rabbits, cats, dogs, pigs, cattle and monkies, as in men (Murakami et al., 1992), essentially all islets in the pancreas were intralobular in location and usually emitted the portal vessels only. In the mouse and rabbit, as in the rat (Murakami and Fujita, 1992), the islet received afferent vessels in its superficial aspect and issued efferent vessels from its deep aspect. In the Formosan monkey, as previously reported in the rhesus monkey (Fujita and murakami, 1973), the afferent vessels usually ran deep into the islet which emitted vessels from its superficial aspect. In other animals examined in this study, as in humans (Murakami et al., 1992), no consistent rule concerning the microcirculatory pattern within the islet could be determined.

Animals↗