Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Inositol Phosphates”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 271 records · Page 15Linked to original sources
PubMed1994

Eicosanoid and inositol phosphate response to platelet-activating factor (PAF) and to a PAF antagonist in rat astroglial cells.

The effects of different concentrations of exogenous platelet-activating factor (PAF) on the formation of arachidonic acid-cyclooxygenase metabolites and on the production of inositol phosphates have been investigated in a primary culture of rat astroglial cells. The cells were used at confluence and the purity was checked by immunostaining of the culture with specific antibodies against glial fibrillary acidic protein. Incubation of the cells with PAF (range 10(-9) to 10(-6) M) resulted in maximal accumulation of total inositol phosphate (620 +/- 60% increment over basal values, P < 0.001) at the concentration of 10(-8) M, after 1 min of stimulation. Smaller inositol phosphate accumulation occurred at higher concentrations of the agonist and at longer stimulation time. After 1 min of stimulation with PAF, the accumulation of the cyclooxygenase metabolites, thromboxane B2 (630 +/- 58 vs 20 +/- 2 pg/mg protein in non-stimulated samples) and 6-keto-prostaglandin F1 alpha (132 +/- 15 vs 55 +/- 7 pg/mg protein in non-stimulated samples) was also maximal at 10(-8) M concentration of the agonist. When the cultures were stimulated with PAF or Ca(2+)-ionophore after preincubation with equimolar concentration of the PAF inhibitor BN 52021, a significant inhibition in the synthesis of both inositol phosphates and cyclooxygenase metabolites occurred only in the PAF-stimulated cells.

6-Ketoprostaglandin F1 alpha↗
PubMed1989

Effects of in vitro hypoxia on depolarization-stimulated accumulation of inositol phosphates in synaptosomes.

The effects of potassium and in vitro histotoxic hypoxia (i.e. KCN) on phosphatidylinositol turnover in rat cortical synaptosomes were determined. [2-3H] Inositol prelabelled rat synaptosomes were prepared from cerebral cortex slices that had been incubated with [2-3H] inositol. Depolarization with 60 mM KCl increased [2-3H] inositol phosphates in a time dependent manner. Depolarization with 60 mM KCl increased [2-3H] inositol trisphosphate transiently at 5 s. K+ induced rapid formation of [2-3H]-inositol bisphosphate and maintained an elevated level for at least 5 min. K+ stimulated gradual formation of [2-3H] inositol monophosphate with time. One minute of hypoxia enhanced potassium-stimulated [2-3H] inositol bisphosphate formation. However, 30 min of hypoxia impaired potassium-stimulated accumulation of [2-3H] inositol phosphates. The effects of histotoxic hypoxia were all dependent upon calcium in the medium and on K+-depolarization. Thus, hypoxia altered the K+-induced accumulation of inositol phosphates in prelabelled synaptosomes in a time dependent, biphasic manner that was calcium dependent.

Analysis of Variance↗
PubMed1993

Potentiation by cholera toxin of bradykinin-induced inositol phosphate production in the osteoblast-like cell line MC3T3-E1.

Cells of the osteoblastic cell line MC3T3-E1 were shown to contain at least three phosphatidylinositol-specific phospholipase C (PI-PLC) isoenzymes (PLC-beta, PLC-gamma and PLC-delta) by Western blotting analysis with various anti-PLC antibodies. Stimulation of inositol phosphate production in MC3T3-E1 cells by bradykinin (BK) occurred via a GTP-binding protein. Inositol phosphate formation on stimulation by BK was not affected by pretreatment with pertussis toxin, whereas it was potentiated by cholera toxin pretreatment. Elevation of cellular cyclic AMP levels by brief pretreatment with dibutyryl cyclic AMP or forskolin failed to enhance the BK-mediated generation of inositol phosphates, but long-term preincubation with these agents partially mimicked the action of the cholera toxin. Cholera toxin also caused an increase in BK receptor number. Cycloheximide, a protein biosynthesis inhibitor, prevented the potentiating actions of the cholera toxin and the cyclic AMP-elevating agents on BK-induced inositol phosphate production, and also inhibited the increase in BK receptor number. The specific binding of [3H]BK to the whole MC3T3-E1 cells in the presence or absence of cholera toxin was completely inhibited by the B2 BK receptor antagonist D-Arg[Hyp3,Thi5,8,D-Phe7]BK, but not by the B1 BK receptor agonist des-Arg9-BK. These data suggest that the activation of PI-PLC induced by cholera toxin in BK-stimulated MC3T3-E1 cells was caused by an enhancement of the synthesis of BK receptor protein(s), at least part of which was mediated by a sustained increase in the intracellular level of cyclic AMP.

Adenosine Diphosphate Ribose↗
PubMed1993

Alterations of inositol phosphate turnover in striatum of aged rats.

Age-related inositol phosphate turnover in the rat central nervous system was investigated. Higher phospholipase-C activity and drastically higher (almost 2.5-fold) inositol 1,4,5-trisphosphate concentration in the corpus striatum (caudate-putamen) of extremely old (approximately 40 months) female Wistar rats in comparison to the young adult (approximately 3.5 months) rats were observed. Dopamine seems to slightly inhibit total inositol phosphate formation and this effect was antagonized by (-)-sulpiride.

Aging↗
PubMed2004

Desensitization of angiotensin-stimulated inositol phosphate accumulation in human vascular smooth muscle cells.

The effect of angiotensin II treatment on desensitization of phospholipase C (PLC)-mediated inositol phosphate accumulation has not been quantitated in human aortic vascular smooth muscle (HVSM) cells. We determined the angiotensin II pretreatment dose dependency and time course for desensitization of PLC activation in HVSM cells and the effect of protein kinase C (PKC) activators on angiotensin II-mediated inositol phosphate accumulation. Our results with PKC activators and direct G protein stimulators suggest that PKC activation may play a negative feedback role in desensitization of angiotensin II-activated signaling in HVSM cells by modifying the Gq transducer, PLC-beta effector, or related proteins in the signaling pathway. However, neither angiotensin II nor PKC activator affected basal phosphorylation levels of PLC-beta1 or PLC-beta3 in HVSM cells; PLC-beta isoenzymes were shown to be phosphorylated in unstimulated cells independent of PKC inhibition. We suggest that desensitization of G protein-stimulated inositol phosphate accumulation in HVSM differs from other cell types in which phosphorylation of PLC-beta isoenzymes accompanies desensitization.

Angiotensin II↗
PubMed1989

Activation of calcium entry by the tumor promoter thapsigargin in parotid acinar cells. Evidence that an intracellular calcium pool and not an inositol phosphate regulates calcium fluxes at the plasma membrane.

The depletion of an inositol 1, 4,5-trisphosphate-sensitive intracellular Ca2+ pool has been proposed to be the signal for Ca2+ entry in agonist-activated cells. Consistent with this idea, thapsigargin, which releases intracellular Ca2+ without inositol phosphate formation, has been reported to activate Ca2+ entry in certain cells. We now report the effects of thapsigargin on Ca2+ entry in parotid acinar cells. In fura-2-loaded parotid acinar cells, thapsigargin caused a sustained elevation of [Ca2+], but did not increase inositol phosphate formation. In the absence of extracellular Ca2+, the increase in [Ca2+], was transient, suggesting that thapsigargin activates both the release of Ca2+ from intracellular stores and the entry of Ca2+ from the extracellular space. In the absence of extracellular Ca2+, pretreatment with methacholine, an agonist believed to mobilize Ca2+ through the production of inositol 1,4,5-trisphosphate, inhibited but did not completely block the response to thapsigargin; likewise, pretreatment with thapsigargin inhibited the response to methacholine. In permeabilized cells, thapsigargin gradually released Ca2+, whereas inositol 1,4,5-trisphosphate caused a rapid and transient discharge of Ca2+. The simultaneous addition of thapsigargin with inositol 1,4,5-trisphosphate evoked a maximum Ca2+ release similar to that for inositol 1,4,5-trisphosphate alone, but the reuptake seen with inositol 1,4,5-trisphosphate alone was abolished. In intact cells, methacholine and thapsigargin together produced a greater initial release of Ca2+ than either alone, but they were not additive in the sustained phase of Ca2+ mobilization. These results demonstrate that the mechanisms for activation of Ca2+ entry by thapsigargin and methacholine are the same and are consistent with the idea that entry is initiated by the depletion of the intracellular inositol 1,4,5-trisphosphate-sensitive Ca2+ pool. The results also indicate that, in contrast to previously proposed models, Ca2+ entry into agonist-activated cells occurs directly across the plasma membrane to the cytoplasm rather than through a cycle of uptake and release by the intracellular Ca2+ pool.

Animals↗
PubMed1995

Lysophosphatidic acid induces inositol phosphate and calcium signals in exocrine cells from the avian nasal salt gland.

We tested lysophosphatidic acid (LPA) known to induce inositol phosphate generation and calcium signals as well as rearrangements of the cytoskeleton and mitogenic responses in fibroblasts, for its ability to activate phospholipase C in an exocrine cell system, the salt-secreting cells from the avian nasal salt gland. LPA (> 10 nmol/l) caused the generation of inositol phosphates from membrane-bound phosphatidylinositides. The resulting calcium signals resembled those generated upon activation of muscarinic receptors, the physiological stimulus triggering salt secretion in these cells. However, close examination of the LPA-mediated calcium signals revealed that the initial calcium spike induced by high concentrations of LPA (> 10 mumol/l) may contain a component that is not dependent upon generation of inositol (1,4,5)-trisphosphate (Ins(1,4,5)P3) and may result from calcium influx from the extracellular medium induced by LPA in a direct manner. Low concentrations of LPA (< 10 mumol/l), however, induce inositol phosphate generation, Ins(1,4,5)P3-mediated release of calcium from intracellular pools and calcium entry. These effects seem to be mediated by a specific plasma membrane receptor and a G protein transducing the signal to phospholipase C in a pertussis-toxin-insensitive manner. Signaling pathways of the muscarinic receptor and the putative LPA-receptor seem to merge at the G-protein level as indicated by the fact that carbachol and LPA trigger hydrolysis of the same pool of phosphatidylinositol (4,5)-bisphosphate (PIP2) and mobilize calcium from the same intracellular stores.

Acetylcholine↗
PubMed1991

Effect of fluoride, pertussis and cholera toxin on the release of arachidonic acid and the formation of prostaglandin E2, D2, superoxide and inositol phosphates in rat liver macrophages.

Fluoride elicited in liver macrophages a release of arachidonic acid and prostaglandins but not formation of inositol phosphates or superoxide. The effects of fluoride required extracellular calcium and were inhibited by staurosporine and by phorbol ester treatment of the cells. Furthermore, fluoride led to a translocation of protein kinase C from the cytosol to membranes. This indicates that the calcium-dependent protein kinase C is involved in the action of fluoride. Cholera toxin decreased the zymosan-induced release of arachidonic acid and prostaglandins but not of inositol phosphates or superoxide. Pertussis toxin ADP-ribosylated a 41,000 molecular weight membrane protein; enhanced specifically the zymosan-induced formation of prostaglandin(PG)E2 but did not affect the zymosan-induced release of arachidonic acid, PGD2, inositol phosphates or superoxide. These data suggest that activation of phospholipase (PL)A2, phosphoinositide (PI)-specific PLC and NADPH oxidase in liver macrophages is most probably not mediated by activation of guanine nucleotide binding (G)-proteins coupled directly to these enzymes.

Adenosine Diphosphate Ribose↗
PubMed1989

Insulin-like growth factor I stimulates inositol phosphate accumulation, a rise in cytoplasmic free calcium, and proliferation in cultured porcine thyroid cells.

Insulin-like growth factor (IGF-I) stimulates thyroid cell proliferation. Using primary cultured porcine thyroid cells, we studied the intracellular pathways that mediate the action of IGF-I on thyroid cell proliferation. IGF-I stimulates inositol phosphate accumulation, a rise in cytoplasmic free calcium [( Ca2+]i), and cell proliferation. Exposure to IGF-I results in a time- and dose-dependent accumulation of inositol monophosphate, inositol bisphosphate, and inositol trisphosphate. IGF-I also increases [Ca2+]i, measured using fura-2, a fluorescent Ca2+ indicator; the IGF-I-induced [Ca2+]i response occurs immediately, reaches a maximum within 1 min, and then slowly declines. IGF-I stimulates thyroid cell proliferation, stimulates thymidine incorporation, and increases cell numbers. The IGF-I-induced inositol phosphate accumulation and [Ca2+]i response parallel thyroid cell proliferation in a dose-dependent manner; the maximal response is observed at a concentration of 100 ng/ml IGF-I, with half-maximal stimulation at approximately 10 ng/ml. Inositol phosphate accumulation and [Ca2+]i response after IGF-I stimulation may function as intracellular messengers for thyroid cell proliferation. This report may constitute the first demonstration of IGF-I-stimulated inositol phosphate accumulation and [Ca2+]i response in the cells.

Animals↗
PubMed1993

Measurement of receptor-activated phosphoinositide turnover in rat brain: nonequivalence of inositol phosphate and CDP-diacylglycerol formation.

Two methods for the measurement of receptor-activated phosphoinositide turnover were evaluated for their degree of correspondence in slices of rat brain; they involved the Li(+)-dependent accumulations of either [3H]-inositol-labeled inositol phosphates or [3H]cytidine-labeled CDP-diacylglycerol. In contrast to the expectation that the ratio of these two responses would remain approximately constant, varying degrees of correspondence were obtained. The two extremes are exemplified by carbachol, which elicited large increases in both inositol phosphate and CDP-diacylglycerol labeling, and endothelin, which gave a robust inositol phosphate response with little or no accumulation of 3H-CDP-diacylglycerol. No instance of the presence of the latter response in the absence of 3H-inositol phosphate accumulation was observed. Measurement of 3H-CDP-diacylglycerol accumulation thus may add additional insight into the regulation of phosphoinositide turnover and the complex action of Li+.

Animals↗
PubMed1995

Selectivity in inositol phosphate production following pancreatic acinar cell desensitization.

The effects of pancreatic acinar cell desensitization by carbamylcholine (Cch), caerulein (Cae) and the phorbol ester TPA on the production of inositol tris and tetrakisphosphates were studied. In control acini, Cch and Cae caused comparable increases in Ins (1,4,5) P3, Ins (1,3,4,5) P4 and Ins (1,3,4) P3 after 15 sec stimulation. Desensitization by Cch totally inhibited the subsequent production of all three inositol phosphates in response to Cch. Desensitization by Cae allowed a subsequent production of all three inositol phosphates in response to Cae, but with a significant reduction. Desensitization by TPA reduced the production of Ins (1,4,5) P3 in response to Cch and Cae and totally inhibited that of the two other inositides. These data point to phospholipase C as a prime target for the altered production of inositol phosphates.

Animals↗
PubMed1991

Halothane inhibits agonist-induced inositol phosphate and Ca2+ signaling in A7r5 cultured vascular smooth muscle cells.

Halothane, an anesthetic with marked depressant effects on the circulation, was studied for its ability to inhibit inositol phosphate and Ca2+ signaling evoked by the vasoactive hormone arginine vasopressin (AVP) and Ca2+ responses elicited by platelet-derived growth factor and by thapsigargin in cultured A7r5 vascular smooth muscle cells. Changes in apparent [Ca2+]i were measured using the indicator indo-1 and flow cytometry, whereas inositol phosphate levels were determined using myo-[3H]inositol and column chromatography. Preincubation with clinically relevant concentrations of halothane resulted in dose-dependent depression of [Ca2+]i responses evoked on stimulation with AVP. Halothane (2.0%) inhibited the increases in [Ca2+]i by 34-45%. In cells incubated in Ca(2+)-free medium plus 0.5 mM ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, the halothane effect was more marked, with 1.5% halothane inhibiting the responses by approximately 53-61%. However, when Ca2+ influx was stimulated by addition of 5 mM Ca2+ in the continued presence of the agonist, the [Ca2+]i response was inhibited by only 15%, suggesting that release of Ca2+ rather than Ca2+ influx is more sensitive to inhibition by the anesthetic. The effects of halothane on Ca2+ homeostasis are not explained solely by anesthetic-induced depletion of Ca2+ from intracellular stores, because the anesthetic inhibited increases in [Ca2+]i elicited by thapsigargin in cells suspended in Ca(2+)-free medium by only 31%. Halothane inhibited inositol phosphate formation elicited by AVP, suggesting an additional means by which the anesthetic may alter agonist-induced Ca2+ responses. The current results also demonstrate that halothane actions are not specific solely to responses evoked by AVP, which acts via a guanine nucleotide-binding protein-linked signaling pathway, but include responses stimulated by platelet-derived growth factor, an agonist that elevates [Ca2+]i via receptor-latent tyrosine kinase activity. The current results demonstrate that, in vascular smooth muscle cells, halothane alters Ca2+ homeostasis, an action that may underlie the in vivo vasodilator effects of the anesthetic.

Animals↗
PubMed1997

[Alteration of inositol phosphate metabolism in aortic smooth muscle cells from spontaneously hypertensive rat].

The changes of inositol phosphates in quiescent and GTP gamma S activated vascular smooth muscle cells (VSMC) derived from aorta of spontaneously hypertensive rat (SHR) were examined. The inositol phosphate assay revealed that there was no difference in the inositol-1, 4, 5-triphosphate (InsP3) level between the quiescent VSMCs of SHR and that of normotensive Wistar-Kyoto rats (WKY). G-protein activator GTP gamma S caused a dose- and time-dependent increase in the InsP3 level in VSMCs. Activated G proteins increased significantly the InsP3 level in VSMCs of SHR. In conclusion, the highly sensitive reactivity to GTP gamma S in the generation of InsP3 in VSMCs from SHR may be related to the possible abnomality of the number, substructure or function of some components in G proteins, phosphoinositide turnover.

Animals↗
PubMed1991

Inositol phosphates and [Ca2+]i signals in a differentiating exocrine cell.

Generation of inositol phosphates and changes in intracellular Ca2+ concentration [( Ca2+]i) upon muscarinic receptor activation were studied in isolated cells from the nasal salt gland of Anas platyrhynchos, comparing responses in the poorly differentiated cells from ducks drinking only tap water (naive cells) with the more fully differentiated actively secreting cells from ducks drinking 1% NaCl solution for 48 h before the experiment (stressed cells). On stimulation, naive cells showed a rapid five- to sevenfold increase in inositol 1,4,5-trisphosphate [Ins(1,4,5)P3], accompanied by similar changes in Ins(1,3,4,5)P4, whereas both values increased only twofold in stressed cells. [3H]quinuclidinyl benzilate binding experiments revealed that these differences in inositol phosphate production were correlated with differences in the numbers of muscarinic acetylcholine receptors. Continuous recordings of [Ca2+]i revealed that Ca2+ release from intracellular stores upon stimulation was similar in both cell types, but the sustained [Ca2+]i signal (dependent on Ca2+ entry) was three times more pronounced in stressed cells. The results suggest that the adaptive differentiation of salt gland cells is associated with the increased expression of the receptor-mediated Ca2+ entry mechanism. In addition, the high rate of phosphoinositide hydrolysis in naive cells upon receptor activation may have a significance in cell growth, proliferation, and differentiation, which are elements of the development of the salt transport capabilities in these cells.

Animals↗
PubMed1986

Guanine nucleotide-dependent pertussis-toxin-insensitive stimulation of inositol phosphate formation by carbachol in a membrane preparation from human astrocytoma cells.

The efficacy of muscarinic-receptor agonists for stimulation of inositol phosphate formation and Ca2+ mobilization in intact 1321N1 human astrocytoma cells is correlated with their capacity for formation of a GTP-sensitive high-affinity binding complex in membranes from these cells [Evans, Hepler, Masters, Brown & Harden (1985) Biochem. J. 232, 751-757]. These observations prompted the proposal that a guanine nucleotide regulatory protein serves to couple muscarinic receptors to the phospholipase C involved in phosphoinositide hydrolysis in 1321N1 cells. Inositol phosphate (InsP) formation was measured in a cell-free preparation from 1321N1 cells to provide direct support for this idea. The formation of InsP3, InsP2 and InsP1 was increased in a concentration-dependent manner (K0.5 approximately 5 microM) by guanosine 5'-[gamma-thio]triphosphate (GTP[S]) in washed membranes prepared from myo-[3H]inositol-prelabelled 1321N1 cells. Both GTP[S] and guanosine 5'-[beta gamma-imido]triphosphate (p[NH]ppG) stimulated InsP formation by 2-3-fold over control; GTP, GDP and GMP were much less efficacious. Millimolar concentrations of NaF also stimulated the formation of inositol phosphates in membrane preparations from 1321N1 cells. In the presence of 10 microM-GTP[S], the muscarinic cholinergic-receptor agonist carbachol stimulated (K0.5 approximately 10 microM) the formation of InsP above that achieved with GTP[S] alone. The effect of carbachol was completely blocked by atropine. The order of potency of nucleotides for stimulation of InsP formation in the presence of 500 microM-carbachol was GTP[S] greater than p[NH]ppG greater than GTP = GDP. Pertussis toxin, at concentrations that fully ADP-ribosylate and functionally inactivate Gi (the inhibitory guanine nucleotide regulatory protein), had no effect on InsP formation in the presence of GTP[S] or GTP[S] plus carbachol. These data are consistent with the idea that a guanine nucleotide regulatory protein that is not Gi is involved in receptor-mediated stimulation of InsP formation in 1321N1 human astrocytoma cells.

Astrocytoma↗
PubMed1994

Neuropeptide Y potentiates norepinephrine-stimulated inositol phosphate production in the rat tail artery.

We have recently demonstrated the ability of neuropeptide Y to augment norepinephrine-stimulated contractions of the rat tail artery. In [3H]myo-inositol labeled segments of rat tail artery, addition of a concentration of neuropeptide Y (10(-8) M) known to potentiate contractile responses produced no effect on the basal accumulation of [3H]inositol phosphates. Addition of norepinephrine (10(-7) M), however, resulted in the rapid accumulation of [3H]inositol 1,4,5-trisphosphate to 325 +/- 29% of basal levels within 5 min. This effect of norepinephrine on [3H]inositol 1,4,5-trisphosphate accumulation was mirrored by significant increases (274 +/- 25% of control) in total [3H]inositol phosphates. When neuropeptide Y and norepinephrine were added together, the norepinephrine-stimulated increase in [3H]inositol 1,4,5-trisphosphate was significantly augmented (467 +/- 35% of control) as was the increase measured in total [3H]inositol phosphates (366 +/- 21% of control). We suggest that the ability of neuropeptide Y to augment norepinephrine-induced contraction of the rat tail artery is due, at least in part, to the ability of neuropeptide Y to increase norepinephrine-stimulated phospholipase C activity in vascular smooth muscle and may represent the mechanism of action of this co-neurotransmitter in vivo.

Animals↗
PubMed1993

Specific oxytocin agonist stimulates prolactin release but has no effect on inositol phosphate accumulation in isolated rat anterior pituitary cells.

We have previously characterized specific oxytocin receptors in the rat anterior pituitary gland, using a highly selective oxytocin receptor antagonist as radioligand. The aim of the present study was to examine whether occupation of these receptors by oxytocin produces a stimulation of prolactin release and a rise in the accumulation of total inositol phosphates in the rat adenohypophysis. Anterior pituitary cells harvested from randomly cycling and diethylstilboestrol (100 micrograms s.c.)-treated rats were perifused with Dulbecco's minimal essential medium at a rate of 0.3 ml/min. Oxytocin and the specific oxytocin agonist [Thr4-Gly7]-oxytocin (TG-OT) both stimulated a significant prolactin release at concentrations of 10(-6) and 10(-7) M. Oestrogen treatment did not affect the response to oxytocin, indicating that there is no straightforward correlation between receptor number and prolactin secretory response in the rat pituitary gland. The involvement of phosphoinositide hydrolysis was investigated in dispersed anterior pituitary cells and uterine tissue from randomly cycling rats. Oxytocin and arginine-vasopressin stimulated a significant (P < 0.05) and dose-related increase in total inositol phosphates, vasopressin being more potent. The specific oxytocin agonist TG-OT had no effect on total inositol phosphate production in pituitary cells, but when tested in uterine tissue it significantly (P < 0.05) stimulated the accumulation of total inositol phosphate at all concentrations tested (10(-5) to 10(-9) M). In conclusion, the data show that oxytocin has prolactin-releasing activity, acting on specific receptors in the anterior pituitary gland.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1997

Calcium ion dependency and the role of inositol phosphates in melatonin-induced encystment of dinoflagellates.

The unicellular eukaryotic dinoflagellates shed their flagella and form a new pellicle cyst wall in response to environmental stress. This encystment process can also be induced by indoleamines such as melatonin and 5-methoxytryptamine. To decipher the complex signaling events which lead to encystment, we have investigated the functional roles of Ca2+ and inositol phosphates in indoleamine-induced encystment of the dinoflagellates Alexandrium catenella and Crypthecodinium cohnii. Pretreatment with EGTA, but not with EDTA, effectively blocked the indoleamine-induced encystment of A. catenella in a dose-dependent manner. Conversely, agents that facilitate the influx of Ca2+ (Bay K 8644, A23187 and ionomycin) dose-dependently induced encystment of A. catenella. Endoplasmic Ca2+-ATPase inhibitors such as thapsigargin and the peptide toxin melittin also induced encystment of A. catenella. These results suggest that an elevation of intracellular [Ca2+] may be involved in the encystment response. In terms of the regulation of phospholipase C, melatonin dose- and time-dependently stimulated the formation of inositol phosphates in C. cohnii. The rank order of potency for several indoleamines to stimulate inositol phosphates formation was 2-iodomelatonin > 5-methoxytryptamine > or = melatonin >> N-acetylserotonin > 5-hydroxytryptamine. This rank order was the same as for the indoleamine-induced encystment of C. cohnii as previously reported. Our results indicate that indoleamine-induced activation of phospholipase C and elevation of intracellular [Ca2+] may be proximal steps in the signal transduction pathway leading to encystment in dinoflagellates. Moreover, this is the first demonstration of the possible involvement of Ca2+ and inositol phosphates as second messengers in dinoflagellates.

5-Methoxytryptamine↗