scholarly journals Is a rise in intracellular concentration of free calcium necessary or sufficient for stimulated cytoskeletal-associated actin?

1986 ◽  
Vol 102 (4) ◽  
pp. 1459-1463 ◽  
Author(s):  
R I Sha'afi ◽  
J Shefcyk ◽  
R Yassin ◽  
T F Molski ◽  
M Volpi ◽  
...  
Keyword(s):  
Intracellular Calcium ◽  
Pertussis Toxin ◽  
Incubation Medium ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Extracellular Calcium ◽  
Intracellular Concentration ◽  
The Other ◽  
Free Calcium ◽  
Ionophore A23187

The addition of the calcium ionophore A23187 to rabbit neutrophils increases the amount of actin associated with the cytoskeleton regardless of the presence or absence of calcium in the incubation medium. In the presence of extracellular calcium, the effect of A23187 is biphasic with respect to concentration. The action of the ionophore is rapid, transient, and is inhibited by pertussis toxin, hyperosmolarity, and quinacrine. On the other hand, the addition of pertussis toxin or hyperosmolarity has small if any, effect on the rise in intracellular calcium produced by A23187. While quinacrine does not affect the fMet-Leu-Phe-induced increase in cytoskeletal actin and the polyphosphoinositide turnover, its addition inhibits completely the stimulated increase in Ca-influx produced by the same stimulus. The results presented here suggest that a rise in the intracellular concentration of free calcium is neither necessary nor sufficient for the stimulated increase in cytoskeletal-associated actin. A possible relationship between the lipid remodeling stimulated by chemoattractants and the increased cytoskeletal actin is discussed.

FMLP-induced enzyme release from neutrophils: a role for intracellular calcium

1983 ◽  
Vol 245 (3) ◽  
pp. C196-C202 ◽  
Author(s):  
D. Chandler ◽  
G. Meusel ◽  
E. Schumaker ◽  
C. Stapleton
Keyword(s):  
Intracellular Calcium ◽  
Cytochalasin B ◽  
Calcium Ionophore ◽  
Cell Ultrastructure ◽  
Calcium Ionophore A23187 ◽  
Extracellular Calcium ◽  
Ionophore A23187 ◽  
Enzyme Release ◽  
Calcium Depletion ◽  
Dose Dependent

The ability of the chemotactic peptide N-formylmethionyl-leucyl-phenylalanine (FMLP) to stimulate beta-glucuronidase release and 45Ca2+ release from rabbit neutrophils was studied. FMLP stimulated enzyme release from cytochalasin B-treated cells either in the presence or the absence of extracellular calcium. Depletion of cell calcium, by exposure to either ethyleneglycol-bis(beta-aminoethylether)-N,N'-tetraacetic acid or the calcium ionophore A23187, blocked the ability of FMLP to stimulate enzyme release and 45Ca2+ release in the absence of extracellular calcium. The ability of A23187 to lower the 45Ca2+ content of neutrophils, to block FMLP-stimulated 45Ca2+ release, and to inhibit FMLP-stimulated enzyme release in the absence of calcium was dose dependent over the same concentration range (10(-8) to 10(-6) M A23187) for all three actions. In contrast, FMLP stimulated enzyme release from A23187-treated cells, provided that extracellular calcium was present. This secretory response was normal as judged by cell ultrastructure and FMLP dose-response relationships. It is concluded that A23187 depletes a pool of intracellular calcium usually released by FMLP and that release of calcium from this pool is necessary for initiation of enzyme secretion in the absence of extracellular calcium.

Cell cycle progression of parthenogenetically activated mouse oocytes to interphase is dependent on the level of internal calcium

1992 ◽  
Vol 103 (2) ◽  
pp. 389-396 ◽  
Author(s):  
C. Vincent ◽  
T.R. Cheek ◽  
M.H. Johnson
Keyword(s):  
Cell Cycle Progression ◽  
Polar Body ◽  
Calcium Ionophore ◽  
Cytosolic Calcium ◽  
Calcium Ionophore A23187 ◽  
Mouse Oocyte ◽  
Free Calcium ◽  
Ionophore A23187 ◽  
Parthenogenetic Activation ◽  
Mouse Oocytes

Nuclear maturation of the mouse oocyte becomes arrested in metaphase of the second meiotic division (MII). Fertilization or parthenogenetic activation induces meiotic completion, chromosomal decondensation and formation of a pronucleus. This completion of meiosis is probably triggered by a transient increase in cytosolic calcium ions. When activated just after ovulation by a low concentration of the calcium ionophore A23187, the majority of the mouse oocytes go through a metaphase to anaphase transition and extrude their second polar body but they do not proceed into interphase; instead their chromatids remain condensed and a microtubular metaphase spindle reforms (metaphase III). However, a high percentage of these oocytes will undergo a true parthenogenetic activation assessed by the formation of a pronucleus, when exposed to a higher concentration of the calcium ionophore. The capacity of the mouse oocyte to pass into metaphase III is lost with increasing time post-ovulation. Direct measurement of intracellular calcium with Fura-2 reveals higher levels of cytosolic calcium in aged oocytes and/or using higher concentrations of calcium ionophore for activation. It is concluded that the internal free calcium level determines the transition to interphase.

scholarly journals Rapid ultrastructural changes in the dense tubular system following platelet activation

Blood ◽  
1992 ◽  
Vol 80 (3) ◽  
pp. 718-723 ◽  
Author(s):  
L Ebbeling ◽  
C Robertson ◽  
A McNicol ◽  
JM Gerrard
Keyword(s):  
Intracellular Calcium ◽  
Platelet Activation ◽  
Calcium Ionophore ◽  
Cytosolic Calcium ◽  
Calcium Ionophore A23187 ◽  
Ultrastructural Changes ◽  
Ionophore A23187 ◽  
Tubular System ◽  
Protein Kinase C Activators ◽  
Morphologic Changes

Abstract The dense tubular system (DTS) functions to regulate platelet activation by sequestering or releasing calcium, similar to the sarcotubules of skeletal muscle. In resting platelets, the DTS exists as thin elongated membranes. Within 10 seconds of the addition of thrombin, platelets show a major ultrastructural change in their DTS: from the thin elongated form to a rounded vesicular form. These morphologic changes were demonstrated with two different stains and two different fixation methods. Platelets exposed to the calcium ionophore A23187 showed the same ultrastructural changes in the DTS. In contrast, the DTS remains in a thin elongated form when platelets are stimulated by the protein kinase C activators phorbol 12-myristate 13-acetate (PMA) and oleoylacetylglycerol (OAG). These morphologic changes may be related to the discharge of calcium from the DTS because this is stimulated by thrombin and A23187, but not by PMA. Preincubation of the platelets with the intracellular calcium chelator 5,5′-dimethyl-bis-(0- aminophenoxy)-ethane-N,N,N′,N tetra acetic acid (BAPTA) largely prevented both the thrombin-induced rise in intracellular calcium and the changes in DTS morphology, suggesting that the changes in DTS morphology are secondary to the increase in cytosolic calcium. The results provide a morphologic correlate to existing biochemical evidence showing that the DTS is involved early during paltelet activation.

scholarly journals The inhibition of neutrophil granule enzyme secretion and chemotaxis by pertussis toxin.

1985 ◽  
Vol 100 (5) ◽  
pp. 1641-1646 ◽  
Author(s):  
E L Becker ◽  
J C Kermode ◽  
P H Naccache ◽  
R Yassin ◽  
M L Marsh ◽  
...  
Keyword(s):  
Pertussis Toxin ◽  
Regulatory Protein ◽  
Calcium Ionophore ◽  
Leukotriene B4 ◽  
Calcium Ionophore A23187 ◽  
Chemotactic Factor ◽  
Target Protein ◽  
Enzyme Secretion ◽  
Ionophore A23187 ◽  
Dependent Inhibition

Pertussis toxin treatment of rabbit peritoneal neutrophils causes a concentration-dependent inhibition of granule enzyme secretion induced by formylmethionyl-leucyl-phenylalanine, C5a, and leukotriene B4. It also inhibits chemotaxis induced by formylmethionyl-leucyl-phenylalanine. The same toxin treatment, however, has no effect on granule enzyme secretion induced by the calcium ionophore A23187 or phorbol 12-myristate 13-acetate. Moreover, pertussis toxin treatment does not affect either the number or affinity of the formylpeptide receptors on the neutrophil nor does it have any effect on the unstimulated levels of cyclic AMP (cAMP) or the transient rise in cAMP induced by chemotactic factor stimulation in these cells. We hypothesize that pertussis toxin, as in other cells, interacts with a GTP binding regulatory protein identical with or analogous to either Ni or transducin which mediates the receptor-induced inhibition or activation of a target protein or proteins required in neutrophil activation. The nature of the target protein is unknown, but it is not the catalytic unit of adenylate cyclase. The target protein acts after binding of chemotactic factor to its receptor in the sequence that leads to the receptor-induced rise in intracellular Ca2+. It does not affect the responses elicited by the direct introduction of calcium into the cells or the activity of protein kinase C.

The relationship between secretion and intracellular free calcium in bovine adrenal chromaffin cells

1984 ◽  
Vol 4 (7) ◽  
pp. 605-611 ◽  
Author(s):  
Robert D. Burgoyne
Keyword(s):  
Chromaffin Cells ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Catecholamine Release ◽  
Intracellular Free Calcium ◽  
Free Calcium ◽  
Ionophore A23187 ◽  
Adrenal Chromaffin Cells ◽  
Adrenal Chromaffin ◽  
The Relationship

The effect of carbamylcholine and the calcium ionophore A23187 on catecholamine release and intracellular free calcium, [Ca2+]i, in bovine adrenal chromaffin cells was determined. At 10−4M carbamylcholine maximal release occurred with an accompanying increase in [Ca2+]i from a basal level of 168 nM to less than 300 nM. An increase in [Ca2+]i of a similar magnitude was found following challenge with 40 nM A23187. However, in this case, no catecholamine release occurred. These results suggest that stimulation of secretion from chromaffin cells by carbamylcholine may involve additional triggers which stimulate secretion at low [Ca2+]i.

The effects of HA compound calcium antagonists on delayed cerebral vasospasm in dogs

1986 ◽  
Vol 65 (1) ◽  
pp. 80-85 ◽  
Author(s):  
Masakazu Takayasu ◽  
Yoshio Suzuki ◽  
Masato Shibuya ◽  
Toshio Asano ◽  
Masahiko Kanamori ◽  
...  
Keyword(s):  
Blood Pressure ◽  
Intracellular Calcium ◽  
Cerebral Vasospasm ◽  
Basilar Artery ◽  
Calcium Antagonists ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Delayed Cerebral Vasospasm ◽  
Arterial Blood

✓ The authors have examined the effects of the HA compounds HA1004(N-(2-guanidinoethyl)-5-isoquino-linesulfonamide) and HA1077(l-(5-isoquinolinesulfonyl)homopiperazine), which are intracellular calcium antagonists, on delayed cerebral vasospasm from subarachnoid hemorrhage (SAH). The modes of action of these compounds were compared with those of the more commonly used calcium entry blockers. Calcium ionophore A23187 (4.8 × 10−6 M)-induced contraction of a canine basilar artery strip was completely antagonized by the HA compounds (10−5 M) but not by the entry-blocking calcium antagonists nicardipine, diltiazem, and verapamil (10−5 M), suggesting that the HA compounds act differently. Delayed cerebral vasospasm was induced by a “two-hemorrhage” canine model. The magnitude of the vasospasm and the effects of the HA compounds were determined angiographically. On SAH Day 7, a significant vasospasm was observed in every dog. The diameter of the basilar artery had diminished to 59% ± 2% (mean ± standard error) of the control value obtained before SAH (on Day 1). The intravenous administration of HA1004 caused a mild dilation of the basilar artery of 10% and 11% at doses of 3 and 10 mg/kg, respectively; however, HA1077 produced a more marked dilation of 19% and 27%, respectively, at the same doses. Both of these drugs lowered mean arterial blood pressure to about 80% and 50% at doses of 3 and 10 mg/kg, respectively. Intracisternal administration of the HA compounds (6 mg) completely reversed cerebral vasospasm without much effect on the blood pressure. The intracellular calcium antagonists of the HA compound group appear to be promising agents for the treatment of intractable cerebral vasospasm.

scholarly journals Involvement of guanine nucleotide binding proteins in neutrophil activation and priming by GM-CSF

Blood ◽  
1989 ◽  
Vol 73 (2) ◽  
pp. 588-591 ◽  
Author(s):  
SR McColl ◽  
C Kreis ◽  
JF DiPersio ◽  
P Borgeat ◽  
PH Naccache
Keyword(s):  
Pertussis Toxin ◽  
Platelet Activating Factor ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Human Neutrophils ◽  
Ionophore A23187 ◽  
Gm Csf ◽  
Surface Receptor ◽  
Pre Treatment ◽  
Guanine Nucleotide Binding

Abstract Pre-incubation of human neutrophils with pertussis toxin significantly inhibited the neutrophil-directed biologic actions of granulocyte- macrophage colony-stimulating factor (GM-CSF) in three separate assays: the induction of c-fos mRNA, the enhancement of both platelet- activating factor-induced mobilization of intracellular calcium, and stimulation of leukotriene synthesis by the calcium ionophore A23187. Cholera toxin did not have an effect on the latter two assays. Pre- treatment of human neutrophils with pertussis toxin did not affect the binding of GM-CSF to its surface receptor. These results provide the first evidence that a pertussis toxin substrate plays an important mediatory role in the mechanism of action of GM-CSF.

Inhibitory effects of calcium ionophore pretreatment of porcine oocytes on polyspermic fertilization

Zygote ◽  
2006 ◽  
Vol 14 (1) ◽  
pp. 17-22 ◽  
Author(s):  
T. Hayashi ◽  
H. Sato ◽  
H. Iwata ◽  
T. Kuwayama ◽  
Y. Monji
Keyword(s):  
Calcium Ionophore ◽  
Inhibitory Effect ◽  
Calcium Ionophore A23187 ◽  
The Other ◽  
Ionophore A23187 ◽  
Inhibitory Effects ◽  
High Concentration ◽  
Maturation Period ◽  
Low Concentration ◽  
Other Hand

The present study examined the inhibitory effects of various pretreatment concentrations (0–100 μM) of the calcium ionophore A23187 on polyspermic fertilization and then examined the effect of the maturation period and the time between calcium ionophore treatment and fertilization on the inhibitory effect of calcium ionophore on polyspermic fertilization. In experiment 1, a high concentration of calcium ionophore (100 μM) increased the rate of activated oocytes, but the rate of fertilization declined. On the other hand, when oocytes were treated with a low concentration of calcium ionophore (10 μM), monospermic fertilization was significantly increased (10 μM; 31.3%) (p < 0.05). In experiment 2, oocytes were cultured for various times (0, 0.5, 3, 6 h) after calcium ionophore treatment (10 μM) before fertilization. The highest rate of monospermic fertilization was detected in the oocytes cultured for 6 h after calcium ionophore treatment before fertilization. In experiments 3 and 4, we examined the effect of the maturation period (40 h or 44 h) on the rate of fertilization and blastulation of oocytes pretreated with calcium ionophore. The treatment of oocytes with calcium ionophore significantly decreased the rate of polyspermic fertilization regardless of the maturation period (44 h: with calcium ionophore 26.25% vs without 78.8%; 40 h: with calcium ionophore 37.5% vs without 77.5%); however, calcium ionophore treatment increased the rates of monospermic fertilization and blastulation of the oocytes matured for 44 h, but not those matured for 40 h. In conclusion, activation with a low concentration of calcium ionophore (10 μM) and a further 6 h of culture before fertilization improved the rate of monospermic fertilization and blastulation.

The role of Src family kinases in starfish egg fertilisation

Zygote ◽  
1999 ◽  
Vol 8 (S1) ◽  
pp. S16-S17 ◽  
Author(s):  
Andrew F. Giusti ◽  
Kathy R. Foltz ◽  
Laurinda. A. Jaffe
Keyword(s):  
Tyrosine Kinase ◽  
Kinase Activity ◽  
Kinase Inhibitors ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Free Calcium ◽  
Ionophore A23187 ◽  
Tyrosine Kinase Activity ◽  
Sh2 Domains ◽  
Upstream Regulator

A common early feature in the activation of all eggs during fertilisation is an increase in the level of intra-cellular free calcium (Ca2+) that, in most species, propagates as a wave across the egg (reviewed in Strieker, 1999). In echinoderms, this Ca2+ release is the result of a signal transduction cascade that requires phospholipase Cγ (PLCγ)-mediated production of inositol trisphosphate (IP3) (Carroll et al., 1997, 1999). PLCγ is most commonly regulated by tyrosine phosphorylation (Rhee & Bae, 1997), indicating that a tyrosine kinase is a likely upstream regulator of PLCγ enzymatic activity at fertilisation. In support of this hypothesis, an increase in tyrosine kinase activity and an increase in tyrosine-phosphorylated proteins at fertilisation has been observed in echinoderm eggs (Satoh & Garbers, 1985; Ciapa & Epel, 1991; Kinsey, 1997). Moreover, the tyrosine kinase inhibitors genistein (Shen et al., 1999) and PP1 (Abassi et al., 2000) have been used to show that in sea urchin eggs a tyrosine kinase activity is required for normal Ca2+ release in response to fertilisation.In eggs of the starfish Asterina miniata, a Src-type tyrosine kinase has been identified as a potential regulator of PLCγ activity at fertilisation (Giusti et al., 1999a). This kinase exhibits a rapid fertilisation-dependent association specifically with the Src Homology 2 (SH2) domains of PLCγ. Moreover, the timing of this association correlates with an increase in the tyrosine kinase activity bound to the PLCγ SH2 domains, and neither the Src kinase nor the associated kinase activity was observed to associate with the PLCγ SH2 domains after treating eggs with the calcium ionophore A23187 (Giusti et al., 1999a). These data identify an egg Src family kinase as a potential upstream regulator of PLCγ during starfish egg fertilisation.

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