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1.
Ca 2+ efflux from the sarcoplasmic reticulum (SR) is routed primarily through SR Ca 2+ release channels (ryanodine receptors, RyRs). When clusters of RyRs are activated by trigger Ca 2+ influx through L-type Ca 2+ channels (dihydropyridine receptors, DHPR), Ca 2+ sparks are observed. Close spatial coupling between DHPRs and RyR clusters and the relative insensitivity of RyRs to be triggered by Ca 2+ together ensure the stability of this positive-feedback system of Ca 2+ amplification. Despite evidence from single channel RyR gating experiments that phosphorylation of RyRs by protein kinase A (PKA) or calcium-calmodulin dependent protein kinase II (CAMK II) causes an increase in the sensitivity of the RyR to be triggered by [Ca 2+] i there is little clear evidence to date showing an increase in Ca 2+ spark rate. Indeed, there is some evidence that the SR Ca 2+ content may be decreased in hyperadrenergic disease states. The question is whether or not these observations are compatible with each other and with the development of arrhythmogenic extrasystoles that can occur under these conditions. Furthermore, the appearance of an increase in the SR Ca 2+ “leak” under these conditions is perplexing. These and related complexities are analyzed and discussed in this report. Using simple mathematical modeling discussed in the context of recent experimental findings, a possible resolution to this paradox is proposed. The resolution depends upon two features of SR function that have not been confirmed directly but are broadly consistent with several lines of indirect evidence: (1) the existence of unclustered or “rogue” RyRs that may respond differently to local [Ca 2+] i in diastole and during the [Ca 2+] i transient; and (2) a decrease in cooperative or coupled gating between clustered RyRs in response to physiologic phosphorylation or hyper-phosphorylation of RyRs in disease states such as heart failure. Taken together, these two features may provide a framework that allows for an improved understanding of cardiac Ca 2+ signaling. 相似文献
2.
Ca 2+ mobilization elicited by simulation with brief pulses of high K + were monitored with confocal laser scanned microscopy in intact, guinea pig cardiac myocytes loaded with the calcium indicator fluo-3. Single wavelength ratioing of fluorescence images obtained after prolonged integration times revealed non-uniformities of intracellular Ca 2+ changes across the cell, suggesting the presence of significant spatial Ca 2+ gradients. Treatment with 20 μM ryanodine, an inhibitor of Ca 2+ release from the SR, and 10 μM verapamil, a calcium channel blocker, reduced by 42% and 76% respectively the changes in [Ca 2+] i elicited by membrane depolarization. The overall spatial distribution of [Ca 2+] i changes appeared unchanged. Ca 2+ transients recorded in the presence of verapamil and ryanodine (about 20% of the size of control responses), diminished in the presence of 50 μM 2-4 Dichlorbenzamil (DCB) or 5 mM nickel, two relatively specific inhibitors of the
exchange mechanism. Conversely, when the reversal potential of the
exchange was shifted to negative potentials by lowering [Na +] 0 or by increasing [Na +] i by treatment with 20 μM monensin, the amplitude of these Ca 2+ transients increased. Ca 2+ transients elicited by membrane depolarization and largely mediated by reverse operation of Na +-Ca 2+ exchange could be recorded in the presence of ryanodine, verapamil and monensin. These findings suggest that in intact guinea pig cardiac cells, Ca 2+ influx through the
exchange mechanism activated by a membrane depolarization in the physiological range can be sufficient to play a significant role in excitation-contraction coupling. 相似文献
3.
We investigated the effect of lysophosphatidic acid (LPA), a bioactive phospholipid, on the response in cytosolic free Ca 2+ concentration ([Ca 2+] i) to mechanical stress in cultured bovine lens epithelial cells. Spritzing of bath solution onto cells as mechanical stress caused marked increase in [Ca 2+] i in the presence of LPA and this increase was concentration-dependent (1–10 μM), whereas neither addition of LPA alone nor the mechanical stress in the absence of LPA affected [Ca 2+] i. The mechanical stress-induced increase in [Ca 2+] i in the presence of LPA was inhibited by removing extracellular Ca 2+ or by addition of Gd 3+, a blocker of mechanosensitive cation channels, but not by nicardipine, thapsigargin, an inhibitor of endoplasmic reticulum-ATPase pump, or U73122, a phospholipase C inhibitor. These results show that LPA sensitises Ca 2+ influx through cation-selective mechanosensitive channels, but does not sensitise Ca 2+ release from intracellular stores, triggered by changes in mechanical stress. On the other hand, phosphatidic acid had less of a sensitising effect than LPA, and neither lysophosphatidylcholine nor chlorpromazine had any effect. Also Ca 2+ mobilising agonists, ATP, histamine and carbachol, did not sensitise Ca 2+ response to the mechanical stress. These results show that LPA sensitises mechanoreceptor-linked response in lens epithelial cells, suggesting that it plays a role in the development of cataracts due to increases in [Ca 2+] i induced by mechanical stress. 相似文献
4.
The effects of PACAPs on [Ca 2+] i were compared to those of carbachol in human neuroblastoma NB-OK-1 cells. PACAP(1–27) and PACAP(1–38) increased [Ca 2+] i in a biphasic manner: a transient rise and a secondary plateau. The transient phase reflected the mobilization of [Ca 2+] i pool(s) via the inositol phosphate pathway. The modest sustained plateau required extracellular Ca 2+. Carbachol also increased [Ca 2+] i in a biphasic manner, but it mobilized intracellular Ca 2+ pool(s) with a higher efficacy than PACAPs, then greatly increased Ca 2+ entry, this being accompanied by a more marked and prolonged elevation of IP 3 and IP 4 than with PACAPs. It is likely that cAMP-mediated phosphorylations due to PACAPs facilitated desensitization at the PACAP receptor-phospholipase C level, so that there was less Ca 2+ handling through PACAP receptors than with muscarinic M 1 receptors. 相似文献
5.
Glucose-induced insulin secretion is pulsatile. We investigated how the triggering pathway (rise in β-cell [Ca 2+] i) and amplifying pathway (greater Ca 2+ efficacy on exocytosis) influence this pulsatility. Repetitive [Ca 2+] i pulses were imposed by high K ++ diazoxide in single mouse islets. Insulin secretion (measured simultaneously) tightly followed [Ca 2+] i changes. Lengthening [Ca 2+] i pulses increased the duration but not the amplitude of insulin pulses. Increasing glucose (5–20 mmol/l) augmented the amplitude of insulin pulses without changing that of [Ca 2+] i pulses. Larger [Ca 2+] i pulses augmented the amplitude of insulin pulses at high, but not low glucose. In conclusion, the amplification pathway ensures amplitude modulation of insulin pulses whose time modulation is achieved by the triggering pathway. 相似文献
6.
We investigated the restoration of [Ca 2+] i in fura-2-loaded human platelets following discharge of internal Ca 2+ stores in the absence of external Ca 2+. After stimulation by thrombin [Ca 2+] i returned from a peak level of 0.6 μM to resting levels within 4 min. When ionomycin discharged the internal stores the recovery was slower with [Ca 2+] i still elevated at around 0.5 μM after 5 min. Thrombin added shortly after ionomycin could accelerate the recovery of [Ca 2+] i and restore resting levels within 5 min, an effect that was mimicked by phorbol-12-myristate-13-acetate (PMA). Since the continued presence of ionomycin precluded reuptake into the internal stores we conclude that thrombin and PMA stimulate Ca 2+ efflux, perhaps via protein kinase C actions on a plasma membrane Ca 2+ pump. 相似文献
7.
Glucose-induced insuline release, glucose-induced rises in intracellular free Ca 2+ concentration ([Ca 2+] i), and voltage-dependent Ca 2+ channel activity were assessed in monolayer cultures of β-vells 3–5 day-old rats. The glucose-stimulated insulin secretory responses and [Ca 2+] i rises were like those in adult rat β-cells rather than fetal rat β-cells. Voltage-dependent Ca 2+ channel antagonists decreased glucose-induced insulin secretion, aborted the [Ca 2+] 2 rise and, like deprivation of extracellular Ca 2+, prevented the glucose-induced rise in [Ca 2+] i when added before the glucose challenge. The presence of nifedipine-sensitive, voltage-dependent Ca 2+ channels was demonstrated directly by measuring Ca 2+ currents using the whole-cell configuration of the patch-clamp technique and indirectly by measuring [Ca 2+] 1 after membrane depolarization by 45 mMm K + or 200 μM tolbutamide. Thus, in cultured β-cells of 3–5 day-old rats the coupling of glucose stimulation to Ca 2+ influx is essentially mature, in contrast to what has been reported for fetal or very early neonatal cells. 相似文献
8.
The effect of clomiphene, an ovulation-inducing agent, on cytosolic free Ca 2+ levels ([Ca 2+] i) in populations of PC3 human prostate cancer cells was explored by using fura-2 as a Ca 2+ indicator. Clomiphene at concentrations between 10-50 μM increased [Ca 2+] i in a concentration-dependent manner. The [Ca 2+] i signal was biphasic with an initial rise and a slow decay. Ca 2+ removal inhibited the Ca 2+ signal by 41%. Adding 3 mM Ca 2+ increased [Ca 2+] i in cells pretreated with clomiphene in Ca 2+-free medium, confirming that clomiphene induced Ca 2+ entry. In Ca 2+-free medium, pretreatment with 50 μM brefeldin A (to permeabilize the Golgi complex), 1 μM thapsigargin (to inhibit the endoplasmic reticulum Ca 2+ pump), and 2 μM carbonylcyanide m-chlorophenylhydrazone (to uncouple mitochondria) inhibited 25% of 50 μM clomiphene-induced store Ca 2+ release. Conversely, pretreatment with 50 μM clomiphene in Ca 2+-free medium abolished the [Ca 2+] i increase induced by brefeldin A, thapsigargin or carbonylcyanide m-chlorophenylhydrazone. The 50 μM clomiphene-induced Ca 2+release was unaltered by inhibiting phospholipase C with 2 μM 1-(6-((17β-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122). Trypan blue exclusion assay suggested that incubation with clomiphene (50 μM) for 2-15 min induced time-dependent decrease in cell viability by 10-50%. Collectively, the results suggest that clomiphene induced [Ca 2+] i increases in PC3 cells by releasing store Ca 2+ from multiple stores in an phospholipase C-independent manner, and by activating Ca 2+ influx; and clomiphene was of mild cytotoxicity. 相似文献
9.
Measurements of Ca 2+ influx and [Ca 2+] i changes in Fura-2/AM-loaded prothoracic glands (PGs) of the silkworm, Bombyx mori, were used to identify Ca 2+ as the actual second messenger of the prothoracicotropic hormone (PTTH) of this insect. Dose-dependent increases of [Ca 2+] i in PG cells were recorded in the presence of recombinant PTTH (rPTTH) within 5 minutes. The rPTTH-mediated increases of [Ca 2+] i levels were dependent on extracellular Ca 2+. They were not blocked by the dihydropyridine derivative, nitrendipine, an antagonist of high-voltage-activated (HVA) Ca 2+ channels, and by bepridil, an antagonist of low-voltage-activated (LVA) Ca 2+ channels. The trivalent cation La 3+, a non-specific blocker of plasma membrane Ca 2+ channels, eliminated the rPTTH-stimulated increase of [Ca 2+] i levels in PG cells and so did amiloride, an inhibitor of T-type Ca 2+ channels. Incubation of PG cells with thapsigargin resulted in an increase of [Ca 2+] i levels, which was also dependent on extracellular Ca 2+ and was quenched by amiloride, suggesting the existence of store-operated plasma membrane Ca 2+ channels, which can also be inhibited by amiloride. Thapsigargin and rPTTH did not operate independently in stimulating increases of [Ca 2+] i levels and one agent’s mediated increase of [Ca 2+] i was eliminated in the presence of the other. TMB-8, an inhibitor of intracellular Ca 2+ release from inositol 1,4,5 trisphosphate (IP 3)-sensitive Ca 2+ stores, blocked the rPTTH-stimulated increases of [Ca 2+] i levels, suggesting an involvement of IP 3 in the initiation of the rPTTH signaling cascade, whereas ryanodine did not influence the rPTTH-stimulated increases of [Ca 2+] i levels. The combined results indicate the presence of a cross-talk mechanism between the [Ca 2+] i levels, filling state of IP 3-sensitive intracellular Ca 2+ stores and the PTTH-receptor’s-mediated Ca 2+ influx. 相似文献
10.
We have studied the effects of cholinegic agonists on the rates of insulin release and the concentrations of diacylglycerol (DAG) and intracellular free Ca 2+ ([Ca 2+] i) in the β-cell line MIN6. Insulin secretion was stimulated by glucose, by glibenclamide and by bombesin. In the presence of glucose, both acetylcholine (ACh) and carbachol (CCh) produced a sustained increase in the rate of insulin release which was blocked by EGTA or verapamil. The DAG content of MIN6 β-cells was not affected by glucose. Both CCh and ACh evoked an increase in DAG which was maximal after 5 min and returned to basal after 30 min; EGTA abolished the cholinergic-induced increased in DAG. ACh caused a transient rise in [Ca 2+] i which was abolished by omission of Ca 2+ or by addition of devapamil. Thus, cholinergic stimulation of β-cell insulin release is associated with changes in both [Ca 2+] i and DAG. The latter change persists longer than the former and activation of protein kinase C and sensitization of the secretory process to Ca 2+ may underlie the prolonged effects of cholinergic agonists on insulin release. However, a secretory response to CCh was still evident after both [Ca 2+] i and DAG had returned to control values suggesting that additional mechanisms may be involved. 相似文献
11.
In cultured pituatary gonadotrophs, gonadotropin-releasing hormone (GnRH) caused dose-dependent and biphasic increases in cytoplasmic calcium concentration ([Ca 2+] i) and LH release. Both extra- and intracellular calcium pools participate in GnRH-induced elevation of [Ca 2+] i and LH secretion. The spike phase of the [Ca 2+] i response represents the primary signal derived predominantly from the rapid mobilization of intracellular Ca 2+. In contrast, the prolonged phase of the Ca 2+ signal depends exclusively on Ca 2+ entry from the extracellular pool. The influx of Ca 2+ occurs partially through dihydropyridine-sensitive calcium channels. Both [Ca 2+] i and LH responses to increasing concentrations of GnRH occur over very similar time scales, suggesting that increasing degrees of receptor occupancy are transduced into amplitude-modulated Ca 2+ responses, which in turn activate exocytosis in a linear manner. However, several lines of evidence indicated the complexity over the relationship between Ca 2+ signaling and LH exocytosis. In contrast to [Ca 2+] i measurements in cell suspension, single cell Ca 2+ measurements revealed the existence of a more complicated pattern of Ca 2+ response to GnRH, with a biphasic response to high agonist doses and prominent oscillatory responses to lower GnRH concentrations, with a log-linear correlation between GnRH dose and the frequency of Ca 2+ spiking. In addition, analysis of the magnitudes of the magnitudes of the [Ca 2+] i and LH responses of gonadotrophs to a wide range of GnRH concentrations in the presence and absence of extracellular Ca 2+, and to K + and phorbol ester stimulation, showed non-linearity between these parameters with amplification of [Ca 2+] i-mediated exocytosis. Studies on cell depleted of protein kinase C under conditions that did not change the LH pool suggested the participation of protein kinase C in this amplication, especially during the plateau phase of the secretory response to GnRH. 相似文献
12.
Stretch of the myocardium influences the shape and amplitude of the intracellular Ca 2+([Ca 2+] i) transient. Under isometric conditions stretch immediately increases myofilament Ca 2+ sensitivity, increasing force production and abbreviating the time course of the [Ca 2+] i transient (the rapid response). Conversely, muscle shortening can prolong the Ca 2+ transient by decreasing myofilament Ca 2+ sensitivity. During the cardiac cycle, increased ventricular dilation may increase myofilament Ca 2+ sensitivity during diastolic filling and the isovolumic phase of systole, but enhance the decrease in myofilament Ca 2+ sensitivity during the systolic shortening of the ejection phase. If stretch is maintained there is a gradual increase in the amplitude of the Ca 2+ transient and force production, which takes several minutes to develop fully (the slow response). The rapid and slow responses have been reported in whole hearts and single myocytes. Here we review stretch-induced changes in [Ca 2+] i and the underlying mechanisms. Myocardial stretch also modifies electrical activity and the opening of stretch-activated channels (SACs) is often used to explain this effect. However, the myocardium has many ionic currents that are regulated by [Ca2+]i and in this review we discuss how stretch-induced changes in [Ca2+]i can influence electrical activity via the modulation of these Ca2+-dependent currents. Our recent work in single ventricular myocytes has shown that axial stretch prolongs the action potential. This effect is sensitive to either SAC blockade by streptomycin or the buffering of [Ca2+]i with BAPTA, suggesting that both SACs and [Ca2+]i are important for the full effects of axial stretch on electrical activity to develop. 相似文献
13.
This study examines the real-time intracellular calcium concentration, [Ca 2+] i, response of canine medial collateral ligament (MCL) and anterior cruciate ligament (ACL) fibroblasts subjected to a fluid-induced shear stress of 25 dynes/cm 2. In experiments using a modified Hanks' Balanced Salt Solution (HBSS) perfusate, both cell types demonstrated a significant increase in peak [Ca 2+] i compared to respective no-flow controls, the response of MCL fibroblasts being nearly 2-fold greater than that of ACL fibroblasts. In studies where the cells were bathed in a medium of HBSS supplemented with 2% newborn bovine serum (NBS) and then introduced to flow with the same medium, ACL fibroblasts responded nearly 3-fold greater than MCL fibroblasts. Neomycin (10 mM), thapsigarigin (1 μM) and Ca 2+-free media supplemented with EGTA (1 mM) were able to inhibit significantly the [Ca 2+] i response to flow with HBSS in both fibroblasts. Thapsigargin also blocked the NBS flow response in both cell types, while neomycin and Ca 2+-free media significantly inhibited the ACL response. Our findings demonstrate that ACL and MCL cells are not the same. These differences may be related to the disparate healing capacity of the ACL and MCL observed clinically. 相似文献
14.
We investigated the effect of newborn bovine serum on the intracellular calcium [Ca 2+] i response of primary cultured bone cells stimulated by fluid flow. As it has been previously established that these cells exhibit [Ca 2+] i responses to fluid flow shear stress in saline media without growth factors or other chemically stimulatory factors, we hypothesized that the addition of serum to the flow medium would enhance the mechanosensitivity of the cells. We examined the effect of a short-term (10–15 min) exposure of the cells to 2 and 10% serum prior to flow stimulation (pretreated) compared to not exposing the cells prior to flow stimulation (unpretreated). The cells were subjected to a well-defined, 90-s flow stimulus with shear stress levels ranging from 0.02 to 3.5 Pa in a laminar flow chamber using a saline medium supplemented with 2 or 10% serum. For pretreatment, the serum concentration was the same from pre-flow to flow exposure. We observed a differential effect in the magnitude of the peak [Ca 2+] i response modulated by the concentration of serum in the pre-flow medium. Additionally, ATP-supplemented flow was examined as a comparison to the serum-supplemented flow and exhibited a similar trend in the peak [Ca 2+] i flow response that was dependent on ATP concentration and pre-flow exposure conditions. These findings demonstrate that under the conditions of this study, chemical agonist exposure can modulate the [Ca 2+] i response in bone cells subjected to fluid flow-induced shear stress. 相似文献
15.
We have investigated the modulation of the intracellular calcium concentration ([Ca 2+] i) in rat C6 glioma cells following their activation by the agonists 5-hydroxytryptamine·HCl (5-HT) and bradykinin, using single cell imaging of [Ca 2+] i with the calcium-sensitive dye Fura-2. The majority of the signals observed involved release of calcium from intracellular stores, and after prolonged application of 5-HT, but not bradykinin, the cells exhibited oscillations in [Ca 2+] i levels. These calcium oscillations were dependent on the presence of extracellular calcium, and were unaffected by the calcium channel antagonists nifedipine and verapamil. Caffeine, which in other cell types is able to release calcium from inositol trisphosphate-insentive stores, had very little effect on [Ca 2+] i levels in C6 cells. On the other hand, bradykinin, although able to elevate [Ca 2+] i probably by acting via the B 2-receptor subtype, was unable to induce any calcium oscillations in these cells. 相似文献
16.
Isolated hepatocytes and the isolated perfused rat liver have been used to study the alterations of cytosolic free Ca 2+ concentration ([Ca 2+] i) produced by 2,5-di(tert-butyl)-l.4-benzohydroquinone (tBuBHQ), a potent inhibitor of hepatic microsomal Ca 2+ sequestration (Moore. G.A., McConkey. D.J., Kass, G.E.N., OBrien, P.J. and Orrenius, S. FEBS Lett., 224, 331-336). (1987). Addition of tBuBHQ to isolated hepatocytes caused a rapid increase in [Ca 2+] i which was similar in magnitude to the [Ca 2+] i elevation induced by the Ca 2+ mobilizing hormone, vasopressin. In contrast with vasopressin which caused a Ca 2+ transient, tBuBHQ elevated [Ca 2+] i to a new steady state that was maintained for up to 15-20min. When vasopressin was administered during the tBuBHQ-induced period of elevated [Ca 2+] i. [Ca 2+] i, rapidly returned to basal levels. Similarly, if vasopressin was administered just prior to tBuBHQ, the resultant tBuBHQ-dependent change in [Ca 2+] i was transient. and not sustained. The hydroquinone mobilized the same intracellular Ca 2+ pool as inositol 1,4,5-trisphosphate. but tBuBHQ did not produce any detectable inositol polyphosphate accumulation. IBuBHQ stimulated glucose release from perifused hepatocytes. mimicking the effect of vasopressin. In the perfused liver, tBuBHQ infusion produced a single, slow and prolonged release of Ca 2+ into the perfusate and inhibition of subsequent vasopressin-induced Ca 2+ effluxes. Inhibition of the response to vasopressin was reversed over time, and closely correlated with the extent of inhibition of both Ca 2+ sequestration and (Ca 2+-Mg 2+)-ATPase activity in microsomes isolated from the isolated perfused liver. The present results are consistent with tBuBHQ inhibiting ATP-dependent Ca 2+ sequestration by a direct effect on the endoplasmic reticular Ca 2+ pump, which results in net Ca 2+ release and elevation of [Ca 2+] i. Furthermore. vasopressin appears to stimulate active removal of increased [Ca 2+] from the hepatocyte cytosol by a mechanism which does not depend on reuptake of Ca 2+ into the endoplasmic reticulum
2,5-Di( tert-butyl) -l,4-benmhydroquinone. calcium. hepatocytes. perfused liver, endoplasmic reticulum 相似文献
17.
Airway myocytes are the primary effectors of airway reactivity which modulates airway resistance and hence ventilation. Stimulation of airway myocytes results in an increase in the cytosolic Ca 2+ concentration ([Ca 2+] i) and the subsequent activation of the contractile apparatus. Many contractile agonists, including acetylcholine, induce [Ca 2+] i increase via Ca 2+ release from the sarcoplasmic reticulum through InsP 3 receptors. Several models have been developed to explain the characteristics of InsP 3-induced [Ca 2+] i responses, in particular Ca 2+ oscillations. The article reviews the modelling of the major structures implicated in intracellular Ca 2+ handling, i.e., InsP 3 receptors, SERCAs, mitochondria and Ca 2+-binding cytosolic proteins. We developed theoretical models specifically dedicated to the airway myocyte which include the major mechanisms responsible for intracellular Ca 2+ handling identified in these cells. These biocomputations pointed out the importance of the relative proportion of InsP 3 receptor isoforms and the respective role of the different mechanisms responsible for cytosolic Ca 2+ clearance in the pattern of [Ca 2+] i variations. We have developed a theoretical model of membrane conductances that predicts the variations in membrane potential and extracellular Ca 2+ influx. Stimulation of this model by simulated increase in [Ca 2+] i predicts membrane depolarisation, but not great enough to trigger a significant opening of voltage-dependant Ca 2+ channels. This may explain why airway contraction induced by cholinergic stimulation does not greatly depend on extracellular calcium. The development of such models of airway myocytes is important for the understanding of the cellular mechanisms of airway reactivity and their possible modulation by pharmacological agents. 相似文献
18.
The effect of five lignans isolated from Hernandia nymphaeifolia on estrogenic compounds (17β-estradiol, tamoxifen and clomiphene)-induced Ca 2+ mobilization in human neutrophils was investigated. The five lignans were epi-yangambin, epi-magnolin, epi-aschantin, deoxypodophyllotoxin and yatein. In Ca 2+–containing medium, the lignans (50–100 μM) inhibited 10 μM 17β-estradiol- and 5 μM tamoxifen-induced increases in intracellular free Ca 2+ levels ([Ca 2+] i) without changing 25 μM clomiphene-induced [Ca 2+] i increase. 17β-estradiol and tamoxifen increased [Ca 2+] i by causing Ca 2+ influx and Ca 2+ release because their responses were partly reduced by removing extracellular Ca 2+. In contrast, clomiphene solely induced Ca 2+ release. The effect of the lignans on these two Ca 2+ movement pathways underlying 17β-estradiol- and tamoxifen-induced [Ca 2+] i increases was explored. All the lignans (50–100 μM) inhibited 10 μM 17β-estradiol-and 5 μM tamoxifen-induced Ca 2+ release, and 17β-estradiol-induced Ca 2+ influx. However, only 100 μM epi-aschantin was able to reduce tamoxifen-induced Ca 2+ influx while the other lignans had no effect. Collectively, this study shows that the lignans altered estrogenic compounds-induced Ca 2+ signaling in human neutrophils in a multiple manner. 相似文献
19.
We have used a continuous spectrofluorimetric method to analyse the role of cytosolic free Ca 2+ ([Ca 2+] i) in the lysosomal enzyme release from the azurophilic granules in human neutrophils stimulated with f-Met-Leu-Phe (fMLP) in the presence of cytochalasin B. Measurements were performed with the β-glucuronidase substrate 4-methylumbelliferyl-β-
-glucuronide. We found that the transient rise in [Ca 2+] i induced by fMLP is a necessary signal to obtain to obtain maximal degranulation. When this Ca 2+ transient is prevented by the Ca 2+ chelator BAPTA, degranulation can still be induced by a stimulated Ca 2+ influx, albeit to a lower extent. We also studied the degranulation process in the neutrophils of a patient with a generalized chemotactic defect. Release of β-glucuronidase from the patient's neutrophils could not be induced despite the occurrence of a normal Ca 2+ response and normal degranulation of specific granules. We conclude that, besides an increase in [Ca 2+] i], an additional signal is required for the fusion of azurophilic granules with the plasma membrane in human neutrophils. 相似文献
20.
The interaction of the novel CCK analogs JMV-180, JMV-320, and JMV-332 with CCK-B/gastrin receptors on small cell lung cancer (SCLC) cells was investigated. JMV-180, JMV-320, and JMV-332 potently inhibited specific binding of 125I-CCK-8 to CCK-B/gastrin receptors expressed on the SCLC cell line NCI-H345 (H345) with IC 50 values of 4.9, 1.8, and 7.0 n M, respectively. JMV-320 and JMV-332 stimulated intracellular calcium ([Ca 2+] i) release in a dose-dependent manner in cells preloaded with indo-1. JMV-180 did not stimulate [Ca 2+] i but inhibited the [Ca 2+] i release elicited by 10 n M CCK-8 in a dose-dependent manner. These data indicate that JMV-320 and JMV-332 function as CCK-B/gastrin receptor agonists while JMV-180 functions as a CCK-B/gastrin receptor antagonist in H345 cells. 相似文献
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