scholarly journals The effect of ionophore A23187 on calcium ion fluxes and α-adrenergic-agonist action in perfused rat liver

1983 ◽  
Vol 214 (2) ◽  
pp. 405-412 ◽  
Author(s):  
P H Reinhart ◽  
W M Taylor ◽  
F L Bygrave
Keyword(s):  
Calcium Ion ◽  
Cellular Level ◽  
Ionophore A23187 ◽  
Respiratory Response ◽  
Adrenergic Agonist ◽  
Experimental Conditions ◽  
Agonist Action ◽  
Total Inhibition ◽  
Hepatic Action ◽  
Alpha Agonist

The effect of ionophore A23187 on cellular Ca2+ fluxes, glycogenolysis and respiration was examined in perfused liver. At low extracellular Ca2+ concentrations (less than 4 microM), A23187 induced the mobilization of intracellular Ca2+ and stimulated the rate of glycogenolysis and respiration. As the extracellular Ca2+ concentration was elevated, biphasic cellular Ca2+ fluxes were observed, with Ca2+ uptake preceding Ca2+ efflux. Under these conditions, both the glycogenolytic response and the respiratory response also became biphasic, allowing the differentiation between the effects of extracellular and intracellular Ca2+. Under all conditions examined the rate of Ca2+ efflux induced by A23187 was much slower than the rate of phenylephrine-induced Ca2+ efflux, although the net amounts of Ca2+ effluxed were similar for both agents. The effect of A23187 on phenylephrine-induced Ca2+ fluxes, glycogenolysis and respiration is dependent on the extracellular Ca2+ concentration. At concentrations of less than 50 microM-Ca2+, A23187 only partially inhibited alpha-agonist action, whereas at 1.3 mM-Ca2+ almost total inhibition was observed. The action of A23187 at the cellular level is complex, dependent on the experimental conditions used, and shows both differences from and similarities to the hepatic action of alpha-adrenergic agonists.

Antidiuretic antagonism and agonism of 1-deamino-pentamethylene-2-d-phenylalanine-4-isoleucine-arginine vasopressin in rats with diabetes insipidus

1987 ◽  
Vol 112 (3) ◽  
pp. 439-442 ◽  
Author(s):  
H. Vilhardt ◽  
S. Lundin
Keyword(s):  
Diabetes Insipidus ◽  
Arginine Vasopressin ◽  
Test Model ◽  
Brattleboro Rats ◽  
Experimental Conditions ◽  
Vasopressin Antagonist ◽  
Agonist Action

ABSTRACT Using implanted minipumps it was shown over a period of 7 days that the vasopressin antagonist, 1-deamino-pentamethylene-2-d-Phe-4-Ile-arginine vasopressin, caused increased diuresis in normal rats and reversed vasopressin- or oxytocin-induced antidiuresis in Brattleboro rats. When the antagonist was infused alone in Brattleboro rats it induced a marked antidiuretic response, indicating that the analogue also possessed agonistic properties. The agonist action could not be demonstrated in anaesthetized, hydrated normal rats. In these animals the analogue behaved as a pure antagonist. It is concluded that analogues which behave as antagonists in one test model may display agonistic properties under different experimental conditions. J. Endocr. (1987) 112, 439–442

scholarly journals Effects of Sex and 17 β-Estradiol on Cardiac Fibroblast Morphology and Signaling Activities In Vitro

Cells ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2564
Author(s):  
Kelsey Watts ◽  
William J. Richardson
Keyword(s):  
Cardiac Fibrosis ◽  
Cardiac Fibroblasts ◽  
Cellular Level ◽  
Cellular Morphology ◽  
Structural Level ◽  
Experimental Conditions ◽  
Male And Female ◽  
Protein Protein Interaction ◽  
Complex Signaling

Several studies have demonstrated estrogen’s cardioprotective abilities in decreasing the fibrotic response of cardiac fibroblasts (CFs). However, the majority of these studies are not sex-specific, and those at the cellular level utilize tissue culture plastic, a substrate with a much higher stiffness than physiological conditions. Understanding the intrinsic differences between male and female CFs under more physiologically “healthy” conditions will help to elucidate the divergences in their complex signaling networks. We aimed to do this by conducting a sex-disaggregated analysis of changes in cellular morphology and relative levels of profibrotic signaling proteins in CFs cultured on 8 kPa stiffness plates with and without 17 β-estradiol (E2). Cyclic immunofluorescent analysis indicated that there was a negligible change in cellular morphology due to sex and E2 treatment and that the differences between male and female CFs occur at a biochemical rather than structural level. Several proteins corresponding to profibrotic activity had various sex-specific responses with and without E2 treatment. Single-cell correlation analysis exhibited varied protein–protein interaction across experimental conditions. These findings demonstrate the need for further research into the dimorphisms of male and female CFs to develop better tailored sex-informed prevention and treatment interventions of cardiac fibrosis.

scholarly journals Purification and properties of a phytase from Candida melibiosica 2491

2018 ◽  
Author(s):  
Danail Georgiev ◽  
Georgi Dobrev ◽  
Stefan Shilev
Keyword(s):  
Molecular Weight ◽  
Copper Ions ◽  
Phytase Activity ◽  
Gel Chromatography ◽  
Experimental Conditions ◽  
Cobalt Ions ◽  
Whole Cells ◽  
Inhibition Effect ◽  
Total Inhibition ◽  
Purification And Properties

Aim: To characterize the enzyme phytase produced by phytase-active Candida melibiosica 2491 for subsecuent use in feed industry. Methods: C. melibiosica 2491 had been selected among 118 strains as the most productive strain of phytase. In present study, the enzyme was first purified through electrophoresis grade in four steps: precipitation with organic solvent, ultrafiltration, gel chromatography and Denaturing gel electrophoresis (SDS–PAGE). Results: Higher levels of purification were obtained using ethanol. The gel chromatography showed an elution maximum at 11-12 fractions that characterize the corresponding one as high-molecular weight phytase. The purification level was found to be 19.5 folds with specific enzyme activity of 2.75 U/mg protein and yield – 19.64 %. Furthermore, the molecular weight of purified phytase was estimated to 35.9 кDa, with optimum of pH – at 4.5 and optimum of temperature at 55 °C. Maximum phytase activity in case of whole cells was found at 50 оС, which was less than using the purified enzyme. It was activated through 5 mM of Ba2+, 10 mM of Mn2+ and K+ ions. Total inhibition effect was achieved from Fe3+, Hg2+ and Zn2+. Copper ions (Cu2+) in concentrations at 5 mM conducted to partial inhibition effect, but at 10 mM the phytase activity was equal to zero. Low inhibition effect was determined in case of cobalt ions (Co2+) at concentrations of 10 mM. The phytase displayed broad sub­strate specificity and the Km for phytate was estimated to be 0.21 mM under the experimental conditions, while Vmax – 19.9 µМ/ml. Conclusion: Although the phytase produced by C. melibiosica 2491 is a promising enzyme to be used successfully in feed production, more investigations are needed to ensure its advantages.

Localization of a pigment-containing structure near the surface of Xenopus eggs which contracts in response to calcium

Development ◽  
1983 ◽  
Vol 76 (1) ◽  
pp. 51-65
Author(s):  
R. W. Merriam ◽  
R. A. Sauterer
Keyword(s):  
Calcium Ion ◽  
Cytochalasin B ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Isolation Medium ◽  
Whole Cells ◽  
Transmission Electron ◽  
Structural Connections ◽  
Contractile Structure

Contractions in surface structures of Xenopus eggs have been induced by application of the calcium ionophore A23187 or calcium ion. Local applications have shown that the contractile structure is present in both animal and vegetal hemispheres. It is, however, much stronger in the animal hemisphere and pigment embedded in it there defines the animal half. The injection of cytochalasin B (CB) into whole cells or the application of the antibiotic to half cells cannot prevent the induced contractions. By experimental means, the contraction of a deeper, pigment-containing structure can be uncoupled from a thin, more superficial and relatively pigment-free layer on the egg surface. By this means it has been possible to establish that the CB-resistant contraction is due, at least partially, to a structurally distinguishable layer subjacent to the outer egg cortex. Scanning and transmission electron microscopy demonstrate a dense grainy matrix near the egg surface in which pigment granules but little yolk are embedded. This structure is much thicker in the pigmented hemisphere. The presence of calcium ions in an isolation medium are shown to cause a loosening or dissolution of the structural connections between the dense, contractile structure near the surface and the cytoskeleton of the endoplasm.

scholarly journals The permeability to calcium of pigeon erythrocyte ‘ghosts’ studied by using the calcium-activated luminescent protein, obelin

1975 ◽  
Vol 152 (2) ◽  
pp. 255-265 ◽  
Author(s):  
Anthony K. Campbell ◽  
Robert L. Dormer
Keyword(s):  
Pyruvate Kinase ◽  
Time Course ◽  
Small Concentration ◽  
Ionophore A23187 ◽  
Erythrocyte Ghosts ◽  
Experimental Conditions ◽  
Bivalent Cation ◽  
Low Concentration ◽  
Triton X ◽  
Obelia Geniculata

1. Obelin, the Ca2+-activated luminescent protein from the hydroid Obelia geniculata, was sealed inside pigeon erythrocyte ‘ghosts’ in order to investigate effects on their permeability of different methods of preparation and of the bivalent cation ionophore A23187. 2. Changes in free Ca2+ within the ‘ghosts’ were studied by following the rate of luminescence of obelin. The possibility that the obelin might have been released from the ‘ghosts’ during an experiment was investigated by studying the release of inulin and pyruvate kinase from the ‘ghosts’. Less than 10% of the inulin or pyruvate kinase sealed within the ‘ghosts’ was released under any of the experimental conditions. 3. Triton X-100 (0.1–10%, v/v) made the ‘ghosts’ highly permeable to Ca2+. In the presence of 1mm-Ca2+ and Triton, 95–100% of the obelin was utilized within 10–20s. 4. A time-course of resealing ‘ghosts’ at 37°C showed that over a period of 90min, the ‘ghosts’ became gradually less permeable to Ca2+. ‘Ghosts’ which remained at 0°C retained only a small concentration of obelin and ATP, and were highly permeable to Ca2+. 5. Erythrocyte ‘ghosts’ resealed for 30min at 20°C rather than 37°C were more permeable to Ca2+, as shown by the fact that 92% of the obelin in the ‘ghosts’ was utilized during the first 60s after the addition of 1mm-Ca2+, as opposed to 44% for ‘ghosts’ resealed at 37°C. 6. Haemolysis at pH6.0 rather than 7.0 resulted in ‘ghosts’ which were highly permeable to Ca2+ after resealing for 60min at 37°C. Of the obelin in the ‘ghosts’, produced by haemolysis at pH6.0, 90% was utilized in the first 60s after the addition of 1mm-Ca2+ compared with 23% for ‘ghosts’ produced at pH7.0. 7. The bivalent cation ionophore A23187 increased the permeability of the ‘ghosts’ to Ca2+. Maximum effects of the ionophore (16μg/ml) were obtained by preincubating the ‘ghosts’ with the ionophore A23187 (16μg/ml) in the presence of a low concentration of Mg2+ and in the absence of Ca2+.

scholarly journals Increased calcium-ion influx is a component of capacitation of spermatozoa

1978 ◽  
Vol 172 (3) ◽  
pp. 549-556 ◽  
Author(s):  
J P Singh ◽  
D F Babcock ◽  
H A Lardy
Keyword(s):  
Calcium Ion ◽  
Time Course ◽  
Acrosome Reaction ◽  
Local Anaesthetics ◽  
Defined Medium ◽  
Ionophore A23187 ◽  
Net Uptake ◽  
Ion Influx ◽  
Triton X

Capacitation (modifications required for gamete fusion) is produced by incubating guinea-pig spermatozoa in vitro in a chemically defined medium. It is shown that during such incubation a net uptake of Ca2+ by the sperm occurs in two distinguishable phases. An initial loose association of Ca2+, possibly to surface sites, is unaffected by agents (Mg2+, inhibitors of mitochondiral function) that prevent or delay the exocytotic spermatozoal acrosome reaction. The time course of a secondary Ca2+ uptake parallels or slightly precedes the time course of the acrosome reaction. This parallelism is maintained during a variety of treatments that either expedite (local anaesthetics, ionophore A23187, Triton X-100) or delay (Mg2+, low external Ca2+) the acrosome reaction. We conclude that the secondary Ca2+ influx described herein apparently serves to link alterations of the spermatozoal membrane to subsequent contractile and secretory components of the capacitation sequence.

scholarly journals Effects of Sex and 17β-Estradiol on Cardiac Fibroblast Morphology and Signaling Activities in vitro

2021 ◽  
Author(s):  
Kelsey Watts ◽  
Will Richardson
Keyword(s):  
Cardiac Fibrosis ◽  
Cardiac Fibroblasts ◽  
Cellular Level ◽  
Cellular Morphology ◽  
Structural Level ◽  
Experimental Conditions ◽  
Male And Female ◽  
Protein Protein Interaction ◽  
Complex Signaling

Several studies have demonstrated estrogen’s cardioprotective abilities in decreasing the fibrotic response of cardiac fibroblasts (CFs). However, the majority of these studies are not sex-specific, and those at the cellular level utilize tissue culture plastic, a substrate that has a stiffness much higher than physiological conditions. Understanding the intrinsic differences between male and female CFs under more physiologically “healthy” conditions will help to elucidate the divergences in their complex signaling networks. We aimed to do this by conducting sex-disaggregated analysis of changes in cellular morphology and relative concentrations of profibrotic signaling proteins in CFs cultured on 8kPa stiffness plates with and without 17-β estradiol (E2). Cyclic immunofluorescent analysis indicated that there is a negligible change in cellular morphology due to sex and E2 treatment and that the differences between male and female CFs are occurring at a biochemical rather than structural level. Several proteins corresponding to profibrotic activity had various sex-specific responses with and without E2 treatment. Single-cell correlation analysis exhibited varied protein-protein interaction across experimental conditions. These findings demonstrate the need for further research into the dimorphisms of male and female CFs to develop better tailored, sex-informed prevention and treatment interventions of cardiac fibrosis.

scholarly journals Liver glucose-6-phosphatase activity is not modulated by physiological intracellular Ca2+ concentrations

1991 ◽  
Vol 275 (3) ◽  
pp. 805-807 ◽  
Author(s):  
R Fulceri ◽  
G Bellomo ◽  
A Gamberucci ◽  
A Benedetti
Keyword(s):  
Phosphatase Activity ◽  
Ionophore A23187 ◽  
Experimental Conditions ◽  
Isolated Liver ◽  
Different Levels

1. In the presence of MgATP and increasing amounts of added Ca2+, isolated liver microsomal vesicles accumulate approx. 10 nmol of Ca2+/mg of protein and buffer ambient free Ca2+ at increasing concentrations (0.22-10.9 microM). Under these experimental conditions, microsomal glucose-6-phosphatase activity is unaffected by the concentration of extravesicular free Ca2+. 2. Different levels of intravesicular Ca2+ were obtained by treating microsomes with the Ca2+ ionophore A23187 and by stimulating active microsomal Ca2+ accumulation with Pi (3 mM). In both instances, microsomal glucose-6-phosphatase activity is unaffected by the level of intravesicular Ca2+.

scholarly journals A 40,000-dalton protein from Dictyostelium discoideum affects assembly properties of actin in a Ca2+-dependent manner.

1982 ◽  
Vol 93 (1) ◽  
pp. 205-210 ◽  
Author(s):  
S S Brown ◽  
K Yamamoto ◽  
J A Spudich
Keyword(s):  
Dictyostelium Discoideum ◽  
Calcium Ion ◽  
Gel Filtration ◽  
Major Effect ◽  
Dependent Manner ◽  
Actin Assembly ◽  
Experimental Conditions ◽  
Native Form ◽  
Proteolytic Fragment ◽  
Peptide Maps

A 40,000-dalton protein that affects the assembly properties of actin in a Ca2+-dependent manner has been purified from Dictyostelium discoideum. Gel filtration chromatography indicates that the native form of this protein is a monomer. A major effect of this protein is to reduce the sedimentability of F-actin in a stoichiometric fashion. Nearly complete loss of sedimentability is observed at ratios of the 40,000-dalton protein to actin of greater than 1:10. At low stoichiometries, this protein can accelerate the rate of actin assembly under certain experimental conditions. These effects of the 40,000-dalton protein on the actin assembly properties described above require calcium ion. The 40,000-dalton protein does not exert its effects by proteolyzing actin. Furthermore, peptide maps demonstrate that this protein is not a proteolytic fragment of actin.

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