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Biomedical subjects

A Baba

Publications and source records attributed to A Baba.

At least 37 records · Page 2Linked to original sources

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

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

Sex difference for tolerance of 5-HT1A receptor-mediated temperature and corticosterone responses in mice.

Repeated treatment with 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) resulted in significant attenuation of 8-OH-DPAT-induced hypothermia and adrenocorticol effect in mice of both sexes, while it did not affect the 8-OH-DPAT-induced decrease in 5-hydroxyindoleacetic acid in the hypothalamus in either sex. The attenuated responses developed more rapidly in female than in male mice, indicating sex differences in the adaptive regulation of the 5-HT1A receptor-mediated responses.

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

Fetal liver T cell receptor gamma/delta+ T cells as cytotoxic T lymphocytes specific for maternal alloantigens.

We have established fetal liver-derived T cell receptor (TCR) gamma/delta+, CD3+ T cell lines that are cytotoxic for maternal T cells. Fetal liver-derived lymphoid progenitors yielded predominantly TCR-gamma/delta+ cell clusters when cultured on fetal bone marrow-derived stromal cells in the presence of a cytokine cocktail under magnetic force. These tightly adherent clusters were cloned by limiting dilution and the resulting cell lines analyzed for phenotype and function. Six of eight TCR-gamma/delta lines from 8-9.5-wk gestation fetuses were V delta 2+ as compared with zero of eight lines from later stages of gestation (10 and 15 wk), where all the lines were V delta 1+. In cytotoxicity assays, these TCR-gamma/delta+, CD3+, CD4-, and CD8+ or CD8- long-term cultured lymphoid cells (LLC) were killer cells active against the class I antigens on maternal T cells. Of the cell lines, the CD8+ TCR-gamma/delta+ LLC had the highest levels of killer activity. Thus fetal liver TCR-gamma/delta+ T cells may play a crucial role in protection against invading maternal T cells generated in the feto-maternal interaction.

Cells, Cultured

Glucose and oxygen deprivation induces a Ca(2+)-mediated decrease in (Na(+)+K+)-ATPase activity in rat brain slices.

Exposure of rat brain cortical slices to a medium lacking in glucose, oxygen or both glucose and oxygen, resulted in a decrease of the tissue ATP content and a reduction of (Na(+)+K+)-ATPase activity in membranes prepared from the slices. These treatments also inhibited partial reactions of (Na(+)+K+)-ATPase such as Na(+)-dependent phosphorylation and K(+)-stimulated phosphatase, as well as specific binding of [3H]ouabain in membranes prepared from the slices. Glucose deprivation and hypoxia decreased (Na(+)+K+)-ATPase activity in the absence of extracellular Ca2+, but the effects were blocked by 1,2-bis(2-amino-phenoxy)ethane-N,N,N',N'-tetraacetic acid tetra-acetomethyl ester (BAPTA-AM), a chelator of intracellular Ca2+. Metabolic inhibitors mimicked the effects of glucose deprivation and hypoxia. The effect of glucose-free hypoxia was dependent on extracellular Ca2+. It was blocked by Mg2+ at high concentration, bepridil or amiloride, but not by voltage-sensitive Ca2+ channel antagonists and glutamate receptor antagonists. None of the drugs tested here, except for dithiothreitol, affected the inhibitory effect of glucose-free hypoxia on the enzyme activity. In contrast to brain (Na(+)+K+)-ATPase, the kidney enzyme was insensitive to glucose and oxygen deprivation and metabolic inhibitors which depleted the tissue ATP.

Animals

Effects of long-term anticonvulsant therapy on copper, zinc, and magnesium in hair and serum of epileptics.

The effects of long-term anticonvulsant therapy on copper (Cu), zinc (Zn), and magnesium (Mg) in the serum and hair were investigated in epileptics. Hair concentrations of Cu in both male and female epileptics, Zn in male epileptics, and Mg in female epileptics were significantly decreased when compared with those of age-matched and gender-matched controls. Hair Cu concentrations were significantly decreased in male epileptics; a significant decrease in hair Mg concentration was observed in female epileptics when compared with schizophrenics. An increased serum Cu concentration was found in female epileptics and a decreased Zn concentration was found in male epileptics. These findings suggest that long-term anticonvulsant therapy could induce alterations in both the metabolism and distribution of Cu, Zn, and Mg.

Adult

Postnatal change in a Ca(2+)-mediated decrease in (Na+ + K+)-ATPase activity in rat brain slices.

The treatment of brain slices from immature rats with veratrine and monensin did not cause any change in (Na+ + K+)-adenosine triphosphatase (ATPase) activity or [3H]ouabain binding in membranes prepared from the slices, though these reagents remarkably stimulated Ca2+ uptake in the slices. Exposure of the slices from adult rats to a glucose-free, hypoxic or both glucose-free and hypoxic medium resulted in a decrease in the enzyme activity, but the enzyme from immature rats was resistant to the conditions.

Animals

[Regulation by chloride ion of astroglial cell functions and morphological transformation].

Recently, several lines of evidence have indicated the important roles of glial cells, especially astrocytes, in the regulation of neuronal functions. The neuron-glia interaction is one of the most important issues in neuroscience, including neuropharmacology. I reviewed the present status and perspectives on the physiologic and pathologic functions of astrocytes in relation to the roles of intracellular Cl-. Astrocytes have different types of Cl- transport systems, such as voltage-sensitive and ligand-gated channels; HCO3(-)-Cl- exchange; and Na+, K+, Cl- cotransport systems. Anion exchange and cotransport systems are responsible for intracellular pH regulation and astrocytic volume regulation, respectively. Especially, astrocytic volume regulation is physiologically important for reducing the concentrations of K+ and glutamate in the extracellular space by their uptake systems. Disturbance of astrocytic volume regulation is expressed as astrocytic swelling, which is usually observed in various brain pathologic states including ischemia. Experimentally, glutamate caused a typical swelling of astrocytes in culture by Cl- and Ca(++)-dependent processes. Glutamate-induced swelling is qualitatively different from reversible swelling induced by hypoosmotic medium. Recently, we found that Cl- is intracellular factor for modulating the receptor-adenylate cyclase system in brain slices. Similarly, the receptor- and forskolin-stimulated adenylate cyclase of astrocytes showed a clear Cl- dependence. This was functionally confirmed by astrocytic morphological transformation induced by the cyclic AMP system.

Astrocytes

Effect of dibutyryl cyclic AMP-treatment on prostaglandin F2 alpha-stimulated phosphoinositide hydrolysis in cultured rat astrocytes.

Dibutyryl cyclic AMP (dBcAMP)-treatment of cultured rat astrocytes induced changes in astrocyte morphology followed by the potentiation of prostaglandin F2 alpha (PGF2 alpha)-stimulated phosphoinositide (PI) hydrolysis. The potentiation was observed in astrocytes of the cerebral cortex, cerebellum, and hippocampus. The dBcAMP-treatment induced agonists-specific changes in PI hydrolysis; e.g., the potentiation of norepinephrine-effect and the reduction of the carbachol-effect. Coincubations of carbachol or norepinephrine with PGF2 alpha produced additive responses.

Animals