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Biomedical subjects

A Baba

Publications and source records attributed to A Baba.

At least 163 records · Page 9Linked to original sources

L-glutamate-stimulated taurine release from rat cerebral cultured astrocytes.

We characterized L-glutamate-stimulated taurine release from cultured astrocytes prepared from rat cerebrum. L-glutamate (0.5 mM) stimulated release of 3H-labeled and endogenous taurine, where the rate of release reached maximum in 40 min. L-glutamate increased astrocytic volume [3H-O-methyl-D-glucose (3H-OMG) space] with a similar time course to 3H-taurine release. Quisqualate, L-aspartate, DL-homocysteate, and L-cysteate increased both astrocytic 3H-OMG space and 3H-taurine release from cultured astrocytes, while kainate (1 mM) stimulated 3H-taurine release without affecting astrocytic volume. N-methyl-D-aspartate had no effect on 3H-taurine release and astrocytic volume. Treatment of astrocytes with dibutyryl cAMP reduced the effect of kainate on 3H-taurine release. L-glutamate-stimulated 3H-taurine release was attenuated by removal of extracellular Cl- and in hyperosmotic medium, which prevented L-glutamate-induced increase in 3H-OMG space of cultured astrocytes. These results indicate that L-glutamate stimulates taurine release from astrocytes through swelling-triggered mechanisms and that kainate causes the release through volume-independent mechanisms.

Animals↗

Identification and characterization of parathyroid hormone/parathyroid hormone-related peptide receptor in cultured astrocytes.

Parathyroid hormone (PTH)/parathyroid hormone-related peptide (PTHrP) receptor expressed in rat cultured cortical type I astrocytes was identified and characterized. Northern blot analysis demonstrated that the PTH/PTHrP receptor mRNA is expressed in astrocytes. A human PTH fragment, PTH [1-34], stimulated adenylate cyclase with the EC50 value of 3 nM, but not phospholipase C. PTH [1-34] changed the morphology of protoplasmic type astrocytes into process-bearing ones. Thus, type I astrocytes possess PTH/PTHrP receptor that is coupled to an adenylate cyclase-activating system but not to phospholipase C.

Adenylyl Cyclases↗

Corticotropin-releasing factor stimulates Ca2+ influx in cultured rat astrocytes.

Corticotropin-releasing factor (CRF) increased intracellular Ca2+ concentration in single astrocytes. The effect in increasing intracellular Ca2+ was not observed in Ca2(+)-free solution. Furthermore, CRF at concentrations more than 10 nM stimulated 45Ca2+ uptake in cultured rat astrocytes. The action was blocked by alpha-helical CRF(9-41) in a competitive manner, but not by nifedipine and 3,4-dichlorobenzamil. On the other hand, CRF did not stimulate cAMP formation, cGMP formation and phosphoinositide hydrolysis in astrocytes. These results indicate that CRF increases Ca2+ influx via an activation of CRF receptors in a cAMP-independent mechanism in cultured astrocytes.

1-Methyl-3-isobutylxanthine↗

Cultured rat astrocytes possess Na(+)-Ca2+ exchanger.

Na(+)-Ca2+ exchange activity in its reverse mode was demonstrated in cultured rat astrocytes. Combination of ouabain (1 mM) and monensin (20 microM) caused a marked increase in 45Ca2+ uptake in astrocytes. 45Ca2+ uptake was also stimulated by lowering the external Na+ concentration. Ouabain plus monensin-stimulated 45Ca2+ uptake was blocked by 3,4-dichlorobenzamil (IC50, 16 microM), an inhibitor of Na(+)-Ca2+ exchanger, but not by nifedipine (0.1 microM). The stimulated-45Ca2+ uptake was observed even in K(+)-free medium, and external K+ at 5-10 mM caused a 2.2-fold increase in the uptake. Microspectrofluorimetry using the Ca(2+)-sensitive dye fura-2 showed that ouabain plus monensin increased intracellular Ca2+ concentration in single astrocytes. The Ca2+ signal was dependent on external Ca2+ (EC50, 1.4 mM), and blocked by 20 microM 3,4-dichlorobenzamil, but not by Ca2+ channel blockers (Cd2+, 20 microM; Ni2+, 100 microM). Antiserum of cardiac Na(+)-Ca2+ exchanger recognized 160 and 120-135 kDa proteins on SDS-polyacrylamide gel electrophoresis of astrocyte homogenate. Northern blot analysis revealed the presence of mRNA for the exchanger protein in astrocytes. These findings indicate that Na(+)-Ca2+ exchanger which is modulated by K+ is present in cultured rat astrocytes.

Amiloride↗

Desensitization of prostaglandin F2 alpha receptor-mediated phosphoinositide hydrolysis in cultured rat astrocytes.

Desensitization of prostaglandin (PG) F2 alpha receptor-mediated phosphoinositide (PI) hydrolysis was investigated in cultured rat astrocytes. Prolonged exposure of astrocytes differentiated by dibutyryl cyclic AMP-treatment to PGF2 alpha caused the desensitization of subsequent PGF2 alpha-induced PI hydrolysis. The desensitization was time- and PGF2 alpha dose-dependent; maximal decrease in the PI hydrolysis was observed after exposure to 10 microM PGF2 alpha for 4 h and the degree of the desensitization was 31.7 +/- 2.7% of control. Pretreatment with either PGD2 or PGE2 also induced the desensitization of subsequent PGF2 alpha-stimulated PI hydrolysis and conversely pretreatment of PGF2 alpha decreased the PI responses to PGD2 and PGE2. The desensitization prevented by phloretin and was reversible upon removal of the agonist. Protein synthesis inhibitors blocked the recovery of the desensitization. Treatment of the cells with phorbol 12-myristate 13-acetate had no effect on the desensitization. These results suggest that prolonged exposure of the astrocytes to PGF2 alpha caused the desensitization of the receptors.

Animals↗

Tunicamycin inhibits prostaglandin F2 alpha receptor-mediated phosphoinositide hydrolysis in cultured rat astrocytes.

Effect of tunicamycin, an inhibitor of N-linked glycosylation, on prostaglandin (PG) F2 alpha-stimulated phosphoinositide (PI) hydrolysis was examined in cultured rat astrocytes. Pretreatment of cultured astrocytes with tunicamycin (25-250 ng/ml) inhibited subsequent PGF2 alpha (1 microM)-stimulated PI-hydrolysis in concentration- and time-dependent manners. The inhibition completely recovered after removal of tunicamycin and re-incubation for 12 h. Tunicamycin pretreatment (100 ng/ml for 12 h) significantly blocked [35S]methionine incorporation into cultured astrocytes, but cell viability was not affected under the condition. Inhibitors of processing of N-linked sugar chains such as bromoconduritol, 1-deoxymannojirimycin, and swainsonine had no effect on PI response to PGF2 alpha. These observations suggest that PGF2 alpha receptor is N-linked glycosylated.

Animals↗

Fluid and protein secretion by the submandibular glands of weanling rats in response to various agonists.

Secretion of fluid and protein by the submandibular glands of 25-day-old rats was investigated by stimulation with 22 sialogogues classified into five categories, four cholinergic, five beta 1-, seven alpha 1- and three alpha 2-adrenergic, and three peptidergic, at optimal doses. For fluid secretion, cholinergic and peptidergic agonists were the most powerful, whereas the beta 1- and alpha 1-adrenoceptor agonists were the most effective for the concentration of protein among the five categories, except for methoxamine. For total output of protein, the beta 1- and alpha 1-adrenoceptor agonists and pilocarpine were the most powerful among the 22 agonists, except for methoxamine and norephedrine. Cholinergic, peptidergic and alpha 2-adrenergic agonists among the five categories were less effective for protein secretion, except for pilocarpine. For the specific activity of esteroprotease, methoxamine and oxymetazoline, as alpha-adrenoceptor agonists, were the most powerful among the 22 agonists. Thus fluid and protein secretion evoked from the submandibular glands of weanling rats in response to a wide variety of agonists are similar to those of adult rats.

Adrenergic Agonists↗

Cloning and expression of a cDNA for rat prostaglandin F2 alpha receptor.

We have cloned a cDNA for rat prostaglandin (PG) F2 alpha receptor from cultured rat astrocytes. The cDNA encodes a polypeptide of 366 amino acids with seven putative transmembrane domains. Specific binding of [3H]PGF2 alpha in membranes of COS-7 cells transfected with the cDNA was displaced with unlabeled PGs in the order of PGF2 alpha > PGD2 > PGE2 > PGI2. In the cDNA-transfected LLC-PK1 cells, PGF2 alpha stimulated phosphoinositide hydrolysis. A significant 4.7-kb mRNA transcript was detected in cultured rat astrocytes and whole brain and pregnant ovary of adult rats by Northern blot analysis.

Amino Acid Sequence↗

Enhanced teratogenic activity of di-n-butyltin diacetate by carbon tetrachloride pretreatment in rats.

Di-n-butyltin diacetate (DBTA) has been shown previously to cause malformations such as cleft mandible, ankyloglossia and fused ribs in rat foetuses after oral treatment of dams on day 8 of gestation. In this study, effects of pretreatment with carbon tetrachloride (CCl4) on the teratogenic activity of DBTA were examined in Wistar rats. Pregnant rats were pretreated sc with CCl4 (0.5 ml/kg) on days 6 and 7 of gestation and were treated orally with DBTA on day 8 of gestation at doses of 0, 4.5, 10 or 22 mg/kg. Caesarean sections were performed on day 20 of gestation, and foetuses were examined for external and skeletal anomalies. Pretreatment with CCl4 enhanced the incidence of external and skeletal malformations caused by DBTA. Concentrations of di-n-butyltin in embryos, maternal liver and maternal blood on day 9 of gestation were increased by CCl4 pretreatment, which inhibited the activity of maternal hepatic microsomal drug-metabolizing enzymes. These results suggest that there is little probability that metabolites of DBTA make a critical contribution to the teratogenicity of DBTA.

Abnormalities, Drug-Induced↗

Endothelins are extracellular signals modulating cytoskeletal actin organization in rat cultured astrocytes.

Effects of endothelin-3 on rapid morphological changes and cytoskeletal actin organization of rat cortical cultured astrocytes were examined. In serum-free medium, treatments with 1 mM dibutyryl cAMP and 5 microM cytochalasin B, an inhibitor of actin polymerization, caused astrocytic morphological changes with cytoplasmic retraction (stellation). Concurrent addition of 1 nM endothelin-3 prevented astrocytic stellation by dibutyryl cAMP and cytochalasin B. The inhibition of endothelin-3 on the astrocytic stellation was dose-dependent, where IC50 and maximal effective dose were about 50 pM and 0.1 nM, respectively. Endothelin-1 and sarafotoxin S6b prevented the cytochalasin B-induced stellation with similar potencies to endothelin-3. Endothelin-3 reversed the stellate morphology of cytochalasin B-treated cells. Sixty minutes after addition of endothelin-3, most cytochalasin B-treated astrocytes lost their apparent distinction between cell body and processes. Treatment with dibutyryl cAMP and cytochalasin B decreased actin content in a 0.5% Triton X-100-insoluble fraction (cytoskeletal fraction) of cultured astrocytes. Subsequent treatments with endothelin-3 for 2 h restored the decreased cytoskeletal actin to that of non-treated cells. Rhodamine-phalloidin staining showed that a prominent structure of organized filamentous actin in protoplasmic astrocytes is stress fibers. The astrocytic stress fibers disappeared after treatment with dibutyryl cAMP and cytochalasin B. Endothelin-3 stimulated reorganization of stress fibers both in the dibutyryl cAMP- and the cytochalasin B-treated astrocytes. These results suggest that endothelins are extracellular signals to regulate cytoskeletal actin organization of astrocytes.

Actins↗

Deficient activity of stimulatory nucleotide-binding regulatory protein in lymphocytes from patients with essential hypertension.

Lymphocytes are widely used as a model for the cardiovascular beta-adrenoceptor-adenylate cyclase system. We evaluated the role of this system in the pathogenesis of hypertension by studying lymphocytes obtained from patients with essential hypertension. Untreated hypertensive patients and normotensive control subjects were studied. The number and affinity of the beta-adrenoceptors were measured by a radioligand binding method with 125I-cyanopindolol. The responses of cyclic adenosine monophosphate (cAMP) to isoproterenol, cholera toxin, and forskolin were also determined. The concentration and affinity of beta-adrenoceptors did not differ significantly in the two groups, nor was a significant difference found in the basal level of cAMP. The effects of isoproterenol on the accumulation of cAMP were reduced in the lymphocytes from the hypertensive compared with the normotensive subjects. There was no significant difference in the effect of forskolin on cAMP accumulation in the two groups. These results indicate that the activity of the stimulatory nucleotide binding regulatory protein (Gs-protein) is reduced in lymphocytes from patients with essential hypertension. This defect of Gs-protein in the lymphocytes may represent a defect of Gs-protein in the cardiovascular system in such patients.

Adenylyl Cyclases↗

Organ-specific and auxin-inducible expression of two tobacco par A-related genes in transgenic plants.

We have isolated four genomic DNA clones that contain the transcription initiation site of the parA gene(s) from a tobacco genomic library by hybridization with the 5' segment of the parA cDNA previously isolated. They were classified into two types on the basis of their nucleotide sequences. Southern blot analysis indicated that two types of clones were respectively derived from the two parental species of tobacco, Nicotiana tomentosiformis and Nicotiana sylvestris. The genes corresponding to these clones were designated as parAt and parAs, respectively, and the parA cDNA clone was shown to code for mRNA from parAt on the basis of its nucleotide sequence. The 5' regions about 400 nucleotides upstream from the transcription initiation sites of the parAt and parAs genomic clones were highly homologous to one another, but regions further upstream showed no significant similarity. The coding sequence of the GUS (beta-glucuronidase) reporter gene was linked to the 5'-upstream regions of parAt and parAs, and the sites of expression of these fusion genes were examined in transgenic tobacco plants. In the absence of auxins, both fusion genes were expressed in capsules at a late stage of seed development, mature seeds, a root apex and a root-hair zone whereas no significant expression was seen in other organs. Their expression was enhanced by 2,4-dichlorophenoxyacetic acid in most of the organs of tobacco. The results show that expression of these genes is regulated by both organ-specific and auxin-inducible mechanisms.

Amino Acid Sequence↗

Angiotensin II subtype 1 receptor modulates epinephrine release from isolated rat adrenal gland.

To elucidate the role of subtypes of the angiotensin (AT) II receptor in epinephrine release from the adrenal gland, the effects of AT II subtype 1 (AT1) receptor and AT II subtype 2 (AT2) receptor antagonists on AT II-induced modulation of epinephrine release were examined. Seven-week-old Wistar rats were used in this study. The left adrenal gland was perfused in a retrograde fashion. The effects of AT II on basal epinephrine release and epinephrine release by electrical stimulation were examined before and after treatment with CV-11974 (CV), an AT1 receptor antagonist, or PD123319 (PD), an AT2 receptor antagonist. AT II increased epinephrine release from the adrenal gland. CV almost completely suppressed AT II-induced increases in epinephrine release. In contrast, PD had no significant effects on AT II-induced increases in epinephrine release. These results suggest that AT II increases epinephrine release from the adrenal medulla via the AT1 receptor.

Adrenal Glands↗

Screening for bacteriuria in Malaysian school children.

Urinary tract infection is the most common of bacterial infections. Screening children for asymptomatic bacteriuria to prevent pyelonephritis and renal scarring is widely recommended. In Malaysia no such attempt has been made to establish the prevalence of asymptomatic bacteriuria. Bacteriuria was screened among 44,816 healthy school children from three different districts in Kelantan. There were 23,132 boys and 21,684 girls. The prevalence of bacteriuria was 0.12% after second screening. Higher prevalence was seen in other reports.

Bacteriuria↗

Inhibitory effect of idazoxan on forskolin-stimulated adenylate cyclase activity through 5-hydroxytryptamine1A receptors.

The binding affinity of the alpha 2-adrenoceptor antagonist idazoxan (CAS 79944-56-2) to 5-hydroxytryptamine1A (5-HT1A) receptors and its effect on the forskolin-stimulated adenylate cyclase activity in the rat hippocampus were examined. Idazoxan had moderate affinity for the 5-HT1A receptors. It inhibited the forskolin-stimulated adenylate cyclase activity to the same extent as 5-HT. The 5-HT1A receptor antagonist spiperone shifted the concentration-response curve of idazoxan to the right in a parallel manner. These results indicate that idazoxan is a 5-HT1A agonist.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

Teratogenic effects of various di-n-butyltins with different anions and butyl(3-hydroxybutyl)tin dilaurate in rats.

In our previous study, di-n-butyltin diacetate has been shown to cause malformations such as cleft mandible, ankyloglossia, fused ribs, etc. in rat fetuses after oral treatment of maternal rats on day 8 of gestation. In this study, teratogenic effects of five di-n-butyltin compounds with different anions and also butyl(3-hydroxybutyl)tin dilaurate (3-OHDBTL) were examined in Wistar rats. Anion groups of these compounds were as follows: diacetate (DBTA), dichloride (DBTC), maleate (DBTM), oxide (DBTO) and dilaurate (DBTL). On day 8 of gestation, pregnant rats were treated orally with these compounds at 80 mumol/kg, or with 3-OHDBTL at 80 or 160 mumol/kg. Cesarean sections were performed on day 20 of gestation and fetuses were examined for their external and skeletal anomalies. Types of the observed malformations were similar to those in the previous study with DBTA except 3-OHDBTL, though the incidence of fetuses with malformations was different. In the DBTC-treated group, skeletal anomalies were predominant to external ones, especially with the higher incidence of fused ribs than in the other groups. In the 3-OHDBTL-treated group, no external and skeletal anomalies were observed at 80 mumol/kg and cleft lower lip and ankyloglossia were observed in only one fetus, but peaked mandible (the tip of lower jaw with acute angle) was specific at 160 mumol/kg. From these results, the di-n-butyl group rather than anionic group was found to be important in the teratogenicity of di-n-butyltin compounds as well as in the other kinds of toxicities and 3-OHDBTL, one of the main metabolites of DBTL, is not the critical substance of teratogenicity because of a very low teratogenic potency.

Animals↗