ET-1-induced bronchoconstriction is mediated via ETB receptor in mice

Nagase, Takahide, Tomoko Aoki, Teruaki Oka, YoshinosukeFukuchi, and Yasuyoshi Ouchi. ET-1-induced bronchoconstriction ismediated via ET_B receptor in mice.J. Appl. Physiol. 83(1): 46-51, 1997.Endothelin (ET)-1 is one of the most potent agonists of airwaysmooth muscle and can act via two different ET receptor subtypes, i.e.,ET_A andET_B. To determine the effects ofET-1 on in vivo pulmonary function and which ET receptors are involved in murine lungs, we investigated 1)the effects of ET and sarafotoxin S6c (S6c), a selectiveET_B agonist, on pulmonary functionand 2) the effects of BQ-123 andBQ-788, specific ET_A- andET_B-receptor antagonists, onET-1-induced bronchoconstriction. ICR mice were anesthetized and mechanically ventilated (frequency = 2.5 Hz, tidalvolume = 8 ml/kg, positive end-expiratory pressure = 3 cmH₂O). Intravenous ET-1, ET-2,and ET-3 increased lung resistance similarly and equipotently, whereasS6c elicited a greater degree of bronchoconstriction. Mice were thenpretreated with saline (Sal), BQ-123 (0.2, 1, and 5 mg/kg), or BQ-788(0.2, 1, and 5 mg/kg) before administration of ET-1(10⁷ mol/kg iv). No dose ofBQ-123 blocked ET-1-induced constriction, whereas pretreatment witheach dose of BQ-788 significantly inhibited ET-1-induced responses.There were significant differences in morphometrically assessed airwayconstriction between Sal and BQ-788 and between BQ-123 and BQ-788,whereas no significant difference was observed between Sal and BQ-123.There were no significant morphometric differences in the airway wallarea among the three groups. These observations suggest that theET_B- but notET_A-receptor subtype may mediatethe changes in murine pulmonary function in response to ET-1. Inaddition, the ET_B-receptorantagonist reduces ET-1-induced airway narrowing by affecting airwaysmooth muscle contraction in mice.

     

The Structure and Characterization of the Surface Protein Layer of a Comamonas-Like Organism

The regular surface layer of a strain of a Comamonas-like organism was examined by electron microscopy. The surface layer protein was easily extracted from the cell surface by a 2.5 M solution of lithium chloride. The protein subunit has a molecular size of 32,000 daltons, but usually forms a large aggregate of more than 1,200,000 daltons. In the extract it formed a regular array of p4 symmetry and was observed to be intimately associated with fragments of lipopolysaccharide. The size of a subunit determined by the negative staining method and the image processing method measured 5.2 × 6.4 nm (width and length), was arranged in a cobblestone-like pattern, and was located in a lattice space measuring 13.0 nm square.     

Effects of anions on ADP-stimulated aggregation of bovine blood platelets

ADP-stimulated aggregation of bovine blood platelets was observed in media containing isotonic potassium salts of various monovalent anions. The aggregation depended on the anion in the medium, the order of aggregation being Cl^?, Br^?>I^?>SCN^?, $\text{ClO}{}_4{}^{\mathrm{?}}$. After 30-min incubation, the extent of aggregation of platelets in Cl^? or Br^? medium was little changed, whereas, that in SCN^? or $\text{ClO}{}_4{}^{\mathrm{?}}$ medium was remarkably decreased. This anion dependency of aggregation may be due to change in the membrane potential.     

A Novel Glial Growth Inhibitory Factor, Gliostatin, Derived from Neurofibroma

Neurofibroma tissue was investigated for the presence of glial growth modulators that would suppress the proliferation of glial cells. A novel endogenous polypeptide inhibitor of proliferation and DNA synthesis in glial cells, gliostatin, was purified from the extracts of neurofibroma by a procedure comprising dye and anion-exchange column chromatography, and HPLC. A monoclonal antibody raised against partially purified gliostatin showed no cross-reactivity with known cytokines, but adsorbed the growth inhibitory activity of gliostatin and immunochemically visualized the putative gliostatin bands on western blot analyses. Although the product showed an apparent M(r) of 100,000 accompanied by an inhibitory activity on gel filtration column chromatography, it migrated at a lower apparent M(r) of 50,000 under the reducing conditions on western blotting, indicating that a homodimeric structure of native gliostatin consisted of 50-kDa subcomponents. Gliostatin was a potent growth inhibitor acting at nanomolar concentrations against all glial tumor cells and glia maturation factor-stimulated astroblasts, but not neuronal cells.     

Increased Expression of β-Amyloid Protein Precursor and Microtubule-Associated Protein τ During the Differentiation of Murine Embryonal Carcinoma Cells

Expression of the genes encoding the beta/A4 amyloid protein precursor (APP) and microtubule-associated protein tau was studied in an embryonal carcinoma cell line (P19) that differentiates in vitro into cholinergic neurons after treatment with retinoic acid. Expression of APP increased 34- (mRNA) and 50-fold (protein) during neuronal differentiation; APP-695 accounted for most of this increase. These remarkable increases in APP expression coincided with a proliferation of neuronal processes and with an increase in content of tau mRNA. Moreover, subsequent decreases in the levels of APP and tau mRNA coincided with the onset of the degeneration of the neuronal processes. Immunocytochemical staining suggested that greater than 85% of the P19-derived neurons are cholinergic and that APP is present in the neuronal processes and cell bodies. These results suggest that APP may play an important role in construction of neuronal networks and neuronal differentiation and also indicate that this embryonal carcinoma cell line provides an ideal model system to investigate biological functions of APP and the roles of APP and tau protein in development of Alzheimer's disease in cholinergic neurons.     

Improved culture conditions for somatic embryogenesis from Asparagus officinalis L. using an aseptic ventilative filter

Summary Calli were induced from the crown of seedlings or lateral bud of young spears of Asparagus officinalis L. on Linsmaier and Skoog's (LS) solid-medium supplemented with 5 M 2,4-dichlorophenoxyacetic acid (2,4-D). Embryogenic callus was selected from induced calli and proliferated in LS liquid medium supplemented with 5 M 2,4-D. Non-vitrified somatic embryos were formed and efficiently developed into club-shaped embryos in LS hormone-free medium with 1 % gelrite in a culture vessel capped with an aseptic ventilative filter. Non-vitrified club-shaped embryos developed into normal plants when transferred to half-strength LS medium without hormones, and 0.8 % agar. Carbon dioxide concentration and moisture content inside the culture vessels were measured, and their effect on embryo development is discussed.     

Protective mechanism of sodium molybdate against the acute toxicity of cadmium in rats. II. Prevention of cytoplasmic acidification

In order to clarify the protective mechanism of sodium molybdate against the acute toxicity of cadmium chloride in rat, the effect of in vivo sodium molybdate pretreatment on the cytotoxic action of cadmium in isolated hepatocytes was studied. The cytosolic pH of hepatocytes isolated from untreated rats immediately decreased with incubation in either neutral Hank's balanced salt solution (HBS), pH 7.4, containing 5 µM cadmium chloride minimum or acidic HBS (pH 7.1, 6.8, 6.5, and 6.2). The presence of 5 µM cadmium in HBS adjusted to pH 7.1 aggravated cytosalic acidification induced by the acidic medium alone. Cell viability of hepatocytes incubated in HBS at pH 6.2 was significantly reduced as compared to that of control cells in HBS at pH 7.4, but the presence of cadmium in the acidic HBS had no aggravating action against such a toxic action of the acidic medium although cellular uptake of the metal in the medium increased, as compared to that in HBS at pH 7.4. Molybdenum pretreatment alleviated cytoplasmic acidification induced by the treatment with HBS at pH 7.4 or 7.1 containing cadmium or by extracellular acid load wothout cadmium. This pretreatment also prevented the loss of cell viability induced by the treatment with HBS at pH 6.2 but could not attenuate that when cadmium was present in the medium.These facts suggest that molybdenum pretreatment alleviated the acute toxicity of cadmium in rat by preventing cytoplasmic acidification caused by the harmful metal.     

Pulse sequences generated by a degenerate analog neuron model

The response characteristics of an electronic neuron model proposed by the authors are investigated. Periodic stimulating pulse sequences with a fixed frequency are applied to the analog neuron model and the response pulse sequences are studied. In the degenerate case, the state transition of the neuron model during one period of the stimulating pulse sequence is described by a first order piecewise linear difference equation with a jump. It is shown that the periodic response pulse sequences of the neuron model belong to a special class of pulse sequences generated by a simple algorithm, and that the relation between the pulse width (or amplitude) of the stimulating pulse and the firing rate of the neuron model takes the form of an extended Cantor function.     

Interactions between the Nucleus and Cytoplasmic Organelles during the Cell Cycle of Euglena gracilis in Synchronized Cultures: II. Associations between the Nucleus and Chloroplasts at an Early Stage of the Cell Cycle under Photoautotrophic Conditions

Chloroplast-nucleus interactions were examined in cells of Euglenagracilis Z synchronized under photoautotrophic conditions. Thechloroplasts were localized near the cell periphery. At an earlystage of the cell cycle, however, some chloroplasts were transientlylocated in the inner space close to the nucleus. Electron microscopyusing serial cell sections revealed that the chloroplast formedprotrusions at several sites, which became associated with thenucleus. The outer membrane of the chloroplast envelope wasin contact, or at least continuous in part, with the outer membraneof the nuclear envelope at the sites of association, and densematerial was present in the chloroplast membrane. A chromosomewas close to each site of the association between these twoorganelles. Most of the chloroplasts including those in associationwith the nucleus were connected by fine bridges. The 4',6-diamidino-2-phenylindole-stainednucleoids in the chloroplast associated with the nucleus appearedto have a thread-like shape. There was another type of chloroplast-nucleusconnection, in which an intervening membranous body was in contactwith the outer part of the nuclear envelope on one side andwith the chloroplast envelope on the other side. ¹ This work was reported at the 48th Annual Meeting of the BotanicalSociety of Japan, Kyoto, October, 1983. (Received June 5, 1984; Accepted November 20, 1984)