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1.
Fernandes AM Landeira-Fernandez AM Souza-Santos P Carvalho-Alves PC Castilho RF 《Neurochemical research》2008,33(9):1749-1758
Excessive activation of NMDA glutamate receptors and the resulting loss of intracellular Ca2+ homeostasis may be lethal (excitotoxic) to neurons. Such excitotoxicity can be induced in vivo by intrastriatal infusion
of quinolinate, as this substance selectively activates NMDA receptors. The aim of the present research was to investigate
whether the in vivo treatment of striatal tissue with quinolinate would lead to an early impairment of sarco/endoplasmic reticulum
Ca2+-ATPase (SERCA) activity or mitochondrial Ca2+ sequestration, two intracellular mechanisms involved in Ca2+ homeostasis and signaling. Sodium quinolinate was infused intrastriatally into adult rats, and 6 h later the brains were
removed and the corpora striata dissected. At this time point, striatal sections stained with Fluoro-Jade, a cellular marker of cell death, showed initial
signs of neuronal degeneration. In addition, SERCA activity decreased 39% in relation to the activity observed in the control
striata. A corresponding decrease of the same magnitude in 45Ca2+ uptake by striatal microsomes was also found in the treated striata. Western blot analysis did not indicate any decrease
in SERCA levels in striatal tissue after quinolinate infusion. Mitochondrial Ca2+ sequestration was still preserved in quinolinate-treated striatal tissue when the assay was carried out in the presence of
physiological concentrations of ATP and Mg2+. These results suggest that impairment of the SERCA function may be an early event in excitotoxicity. 相似文献
2.
In the resting state, the Ca2+ concentration in agonist-sensitive intracellular stores reflects the balance between active uptake of Ca2+, which is mediated by Ca2+-ATPase (SERCA), and passive leakage of Ca2+. The mechanisms underlying such a leakage in cells of the submaxillary salivary gland were not studied. In our experiments,
we examined possible pathways of passive leakage of Ca2+ from the endoplasmic reticulum (ER) of acinar cells obtained from the rat submaxillary salivary gland; direct measurements
of the concentration of Ca2+ in the ER ([Ca2+]ER) using a low-affinity calcium-sensitive dye, mag-fura 2/AM, were performed. The cellular membrane was permeabilized with
the help of β-escin (40 μg/ml); the Ca2+ concentration in the cytoplasm ([Ca2+]
i
) was clamped at its level typical of the resting state (∼100 nM) using an EGTA/Ca2+ buffer. Incubation of permeabilized acinar cells in a calcium-free intracellular milieu, as well as application of thapsigargin,
resulted in complete inhibition of the uptake of Ca2+ with the involvement of SERCA. This effect was observed 1 min after the beginning of superfusion of the cells with the corresponding
solutions and was accompanied by the leakage of Ca2+ from the ER; this is confirmed by a gradual drop in the [Ca2+]ER. Such a leakage of Ca2+ remained unchanged in the presence of thapsigargin, heparin, and ruthenium red; therefore, it is not mediated by SERCA, inositol
1,4,5-trisphosphate-sensitive receptors (InsP3R), or ryanodine receptors (RyRs). At the same time, an antibiotic, puromycin (0.1 to 1.0 mM), which disconnects polypeptides
from the ER-ribosome translocon complex, caused intensification of passive leakage of Ca2+ from the ER. This effect did not depend on the functioning of SERCA, InsP3R, or RyR. Therefore, passive leakage of Ca2+ from the ER in acinar cells of the submaxillary salivary gland is realized through pores of the translocon complex of the
ER membrane.
Neirofiziologiya/Neurophysiology, Vol. 37, No. 4, pp. 339–346, July–August, 2005. 相似文献
3.
Enzymes are able to handle the energy derived from the hydrolysis of phosphate compounds in such a way as to determine the
parcel that is used for work and the fraction that is converted into heat. The sarco/endoplasmic reticulum Ca2+-ATPases (SERCA) is a family of membrane-bound ATPases that are able to transport Ca2+ ion across the membrane using the chemical energy derived from ATP hydrolysis. The heat released during ATP hydrolysis by
SERCA may vary from 10 up to 30 kcal/mol depending on the SERCA isoform used and on whether or not a Ca2+ gradient is formed across the membrane. Drugs such as heparin, dimethyl sulfoxide and the platelet-activating factor (PAF)
are able to modify the fraction of the chemical energy released during ATP hydrolysis that is used for Ca2+ transport and the fraction that is dissipated in the surrounding medium as heat. The thyroid hormone 3,5,3′-triiodo L-thyronine (T3) regulates the expression and function of the thermogenic SERCA isoforms. Modulation of heat production by SERCA might be
one of the mechanisms involved in the increased thermogenesis found in hyperthyroidism. 相似文献
4.
Isolated sarcoplasmic reticulum vesicles in the presence of Mg(2+) and absence of Ca(2+) retain significant ATP hydrolytic activity that can be attributed to the Ca(2+)-ATPase protein. At neutral pH and the presence of 5 mM Mg(2+), the dependence of the hydrolysis rate on a linear ATP concentration scale can be fitted by a single hyperbolic function. MgATP hydrolysis is inhibited by either free Mg(2+) or free ATP. The rate of ATP hydrolysis is not perturbed by vanadate, whereas the rate of p-nitrophenyl phosphate hydrolysis is not altered by a nonhydrolyzable ATP analog. ATP binding affinity at neutral pH and in a Ca(2+)-free medium is increased by Mg(2+) but decreased by vanadate when Mg(2+) is present. It is suggested that MgATP hydrolysis in the absence of Ca(2+) requires some optimal adjustment of the enzyme cytoplasmic domains. The Ca(2+)-independent activity is operative at basal levels of cytoplasmic Ca(2+) or when the Ca(2+) binding transition is impeded. 相似文献
5.
Leopoldo de Meis 《Bioscience reports》2001,21(2):113-137
The sarcoplasmic reticulum of skeletal muscle retains a membrane bound Ca2+-ATPase which is able to interconvert different forms of energy. A part of the chemical energy released during ATP hydrolysis is converted into heat and in the bibliography it is assumed that the amount of heat produced during the hydrolysis of an ATP molecule is always the same, as if the energy released during ATP cleavage were divided in two non-interchangeable parts: one would be converted into heat, and the other used for Ca2+ transport. Data obtained in our laboratory during the past three years indicate that the amount of heat released during the hydrolysis of ATP may vary between 7 and 32 kcal/mol depending on whether or not a transmembrane Ca2+ gradient is formed across the sarcoplasmic reticulum membrane. Drugs such as heparin and dimethyl sulfoxide are able to modify the fraction of the chemical energy released during ATP hydrolysis which is used for Ca2+ transport and the fraction which is dissipated in the surrounding medium as heat. 相似文献
6.
7.
Experiments were carried out on isolated neurons of the thalamic nucleus lateralis dorsalis (LD) from 12-day-old rats. According to the morphological characteristics, LD neurons were classified as relay thalamo-cortical units and interneurons. The concentration of free Ca2+ ions in the cytoplasm ([Ca2+]
i
) was measured by a fluorescent calcium indicator, fura-2AM. Application of 30 mM caffeine caused a transient change in the [Ca2+]
i
in 8 of 15 and in 6 of 11 of the thalamo-cortical units and interneurons under study, respectively. After stimulation of a cell with application of 50 mM KCl, a caffeine-induced increase in the [Ca2+]
i
was observed in all tested neurons. To study the contribution of Ca2+-induced Ca2+ release (CICR) to the calcium transient evoked by depolarization of the neuronal membrane, caffeine in a subthreshold concentration was pre-applied. After 50 mM KCl had been added to the medium following pre-application of 0.5 mM caffeine, the calcium transient amplitude in thalamo-cortical neurons increased by 51 ± 7% (n = 16). In interneurons this effect was not observed (n = 11). The data obtained allow us to hypothesize that CICR contributes to the depolarization-evoked calcium transient only in the relay (thalamo-cortical) neurons. Differences in the pattern of calcium signalling, which were detected in two types of neurons of the thalamic LD, can be a factor determining distinctions in the physiological characteristics of these neurons. 相似文献
8.
Marijn Kuijpers Gaga Kochlamazashvili Alexander Stumpf Dmytro Puchkov Aarti Swaminathan Max Thomas Lucht Eberhard Krause Tanja Maritzen Dietmar Schmitz Volker Haucke 《Neuron》2021,109(2):299-313.e9
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9.
《Molecular membrane biology》2013,30(4):191-201
The uncoupling of Ca2+ transport from ATP hydrolysis in the sarcoplasmic reticulum (Ca2+ + Mg2+)-ATPase by trypsin digestion was re-investigated by comparing ATPase activity with the ability of the enzyme to occlude Eu3+ (a transport parameter) after various tryptic digests. With this method, re-examination of uncoupling by tryptic digest of the ATPase revealed that TD2 cleavage (Arg-198) had no effect on either occlusion or ATPase activity. Digestion past TD2 in the presence of 5 mM Co2+ and at 25°C resulted in the loss of about 70% of the ATPase activity, but no loss of occlusion. Digestion past TD2 in the presence of 5 mM Ca2+, 3 mM ATP, and at 25°C resulted in a partially uncoupled enzyme complex which retained about 50% of the ATPase activity, but completely lost the ability to occlude Eu3+. Digest past TD2 in the presence of 5 mM Ca2+ and 3 mM AMP-PNP. (a non-hydrolyzable ATP analog) at 25°C resulted in no loss of occlusion, thus revealing the absolute requirement of ATP during the digest to eliminate occlusion. From these findings we conclude that uncoupling of Ca2+ transport from ATPase activity is possible by tryptic digestion of the (Ca2+ + Mg2+)-ATPase. Interestingly, only after phosphorylation of the enzyme do the susceptible bond(s) which lead to the loss of occlusion become exposed to trypsin. 相似文献
10.
Takahashi Y Murakami M Watanabe H Hasegawa H Ohba T Munehisa Y Nobori K Ono K Iijima T Ito H 《Biochemical and biophysical research communications》2007,356(1):45-52
Store-operated Ca(2+) entry (SOCE) is a physiologically important process that is triggered by intracellular Ca(2+) depletion. Recently, human Orai1 (the channel-forming subunit) and STIM1 (the calcium sensor) were identified as essential molecules for SOCE. Here, we report the cloning and functional analysis of three murine orthologs of Orai1, termed Orai1, 2, and 3. Among the genes identified, Orai1 contains a distinctive proline- and arginine-rich N-terminal cytoplasmic sequence. Co-expression of STIM1 with Orai1 produced a marked effect on SOCE, while co-expression with Orai2 or Orai3 had little effect. Expression of Orai1 without its N-terminal tail had a marginal effect on SOCE, while chimeric Orai2 containing the Orai1 N-terminus produced a marked increase in SOCE. In addition, a truncated version of Orai1 containing the N-terminus without the pore-forming transmembrane domain had a dominant negative effect on SOCE. These results reveal the essential role of Orai1 and its N-terminal sequence in SOCE. 相似文献
11.
Pannexin channels in ATP release and beyond: An unexpected rendezvous at the endoplasmic reticulum 总被引:1,自引:0,他引:1
Catheleyne D'hondt Raf Ponsaerts Humbert De SmedtMathieu Vinken Elke De VuystMarijke De Bock Nan WangVera Rogiers Luc LeybaertBernard Himpens Geert Bultynck 《Cellular signalling》2011,23(2):305-316
The pannexin (Panx) family of proteins, which is co-expressed with connexins (Cxs) in vertebrates, was found to be a new GJ-forming protein family related to invertebrate innexins. During the past ten years, different studies showed that Panxs mainly form hemichannels in the plasma membrane and mediate paracrine signalling by providing a flux pathway for ions such as Ca2+, for ATP and perhaps for other compounds, in response to physiological and pathological stimuli. Although the physiological role of Panxs as a hemichannel was questioned, there is increasing evidence that Panx play a role in vasodilatation, initiation of inflammatory responses, ischemic death of neurons, epilepsy and in tumor suppression. Moreover, it is intriguing that Panxs may also function at the endoplasmic reticulum (ER) as intracellular Ca2+-leak channel and may be involved in ER-related functions. Although the physiological significance and meaning of such Panx-regulated intracellular Ca2+ leak requires further exploration, this functional property places Panx at the centre of many physiological and pathophysiological processes, given the fundamental role of intracellular Ca2+ homeostasis and dynamics in a plethora of physiological processes. In this review, we therefore want to focus on Panx as channels at the plasma membrane and at the ER membranes with a particular emphasis on the potential implications of the latter in intracellular Ca2+ signalling. 相似文献
12.
NMDA受体通道的结构与功能 总被引:6,自引:0,他引:6
近来用分子克隆方法对N-甲基-门冬氨酸受体(NMDA受体)通道的分子结构进行了广泛的研究.这些研究清楚地显示了NMDA受体通道的分子多样性,为NMDA受体通道的在体功能多样性提供了基础.已获得的克隆为研究这些受体通道分布和生理作用提供了有价值的工具. 相似文献
13.
We showed that 5 M acetylcholine (ACh) and 100 M norepinephrine (NE) cause increases in the total Ca2+ content in acinar cells by 30 and 87% and in the exocytosis intensity by 15 and 20%, respectively. Application of 5 M ACh and 100 M NE increased the free cytosolic Ca2+ concentration ([Ca2+]
i
) by 87 ± 2 and 140 ± 7 nM, respectively. Application of ACh and NE in a Ca2+-free external solution caused a [Ca2+]
i
increase that was 40 and 67% lower than in physiological solution. We postulate that the exocytosis developing upon neural stimulation of the gland results from generation of Ca2+ transients that are spreading from the basal to the apical region of the exocrine cell, where secretory granules are concentrated. 相似文献
14.
The subcellular distribution of endoplasmic reticulum proteins (IP3R1 and RYR), plasma membrane(PM) proteins (mGluR1 and PMCA Ca2+-pump), and scaffolding proteins, such as Homer 1b/c, was assessed by laser scanning confocal microscopy of rat cerebellum parasagittal sections. There appeared to be two classes of Ca2+ stores, nonjunctional Ca2+ stores and junctional Ca2+ stores, possibly referable to central cisternae/tubules and sub-PM cisternae, respectively, in soma, dendrites, and dendritic spines. Only some IP3R1s appeared to be part of multimeric, junctional Ca2+ signaling networks, whose composition is shown to include PMCA, mGluR1, Homer 1b/c and, not always, RYR1. 相似文献
15.
Rough endoplasmic reticulum (RER) prepared from bulk-isolated neuronal perikarya of rat brain cortex of different postnatal ages was found to be active in vectorial discharge of nascent proteins through the membrane; this activity increased with the increasing age of animals and reached maximal values in adults. RER isolated from whole cortical tissue (containing all cell types) exhibited vectorial release only up to 18 days of age; the preparation from adult animals was essentially devoid of secretory activity. Controlled proteolysis of various preparations suggested that in neuronal RER of 8-day-old rats the proportion of nascent proteins operationally retained in the intravesicular space was about twice that retained by cortical preparations. For the purpose of comparison, these parameters were studied also in liver RER. 相似文献
16.
杨树顶芽细胞内质网与其他膜系统的结构联系及其在休眠过程中的变化 总被引:8,自引:0,他引:8
杨树(Populus deltoides Bartr.ex Marsh)顶芽分生组织细胞经一种改良的高锰酸钾固定法固定后,显示出一种十分清晰的内膜结构,尤其展现了内质网与其他膜系统存在一种结构上的密切联系。一些与核膜相连接的内质网伸展到细胞质中与线粒体、质体及高尔基体发生联系,可延伸到质膜。还有些内质网的一端与一个细胞的核膜相连结,其另一端穿过胞间连丝与邻近的另一个细胞的核膜相连结,在两个相邻的细 相似文献
17.
Yuh-Jiin Jong Adrian Sheldon Guo H. Zhang Naomi Kraus-Friedmann 《The Journal of membrane biology》1990,118(1):49-53
Summary The Ca2+-ATPase from rat liver microsomes has been solubilized in Triton X-100 and purified to homogeneity by ficollsucrose treatment, column chromatography with agarose-hexane adenosine 5-triphosphate Type 2, and high pressure liquid chromatography (HPLC). The purified enzyme obtained by this sequential procedure exhibited a 183-fold increase in specific activity. After ficoll-sucrose treatment, the activity of the Ca2+-ATPase was stable for at least two weeks when stored at –70°C. In SDS-polyacrylamide gels, several fractions from HPLC chromatography showed a single band at a position corresponding to a molecular weight of about 107 kDa. This value is consistent with the molecular weight of the phosphoenzyme intermediate of endoplasmic reticulum (ER) Ca2+-ATPase. Further characterization of the ER Ca2+-ATPase was performed by western immunoblots. Antiserum raised against the 100-kDa sarcoplasmic reticulum (SR) Ca2+-ATPase cross-reacted with the purified Ca2+-ATPase from rat liver ER membranes. 相似文献
18.
Vladimir Parpura Vladimir Grubiši?Alexei Verkhratsky 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》2011,1813(5):984-991
Astrocytes can exocytotically release the gliotransmitter glutamate from vesicular compartments. Increased cytosolic Ca2+ concentration is necessary and sufficient for this process. The predominant source of Ca2+ for exocytosis in astrocytes resides within the endoplasmic reticulum (ER). Inositol 1,4,5-trisphosphate and ryanodine receptors of the ER provide a conduit for the release of Ca2+ to the cytosol. The ER store is (re)filled by the store-specific Ca2+-ATPase. Ultimately, the depleted ER is replenished by Ca2+ which enters from the extracellular space to the cytosol via store-operated Ca2+ entry; the TRPC1 protein has been implicated in this part of the astrocytic exocytotic process. Voltage-gated Ca2+ channels and plasma membrane Na+/Ca2+ exchangers are additional means for cytosolic Ca2+ entry. Cytosolic Ca2+ levels can be modulated by mitochondria, which can take up cytosolic Ca2+ via the Ca2+ uniporter and release Ca2+ into cytosol via the mitochondrial Na+/Ca2+ exchanger, as well as by the formation of the mitochondrial permeability transition pore. The interplay between various Ca2+ sources generates cytosolic Ca2+ dynamics that can drive Ca2+-dependent exocytotic release of glutamate from astrocytes. An understanding of this process in vivo will reveal some of the astrocytic functions in health and disease of the brain. This article is part of a Special Issue entitled: 11th European Symposium on Calcium. 相似文献
19.
Emeran A. Mayer Anatoly Kodner Xiao Ping Sun Jonathan Wilkes David Scott George Sachs 《The Journal of membrane biology》1992,125(2):107-118
Summary Intracellular calcium [Ca2+]
i
measurements in cell suspension of gastrointestinal myocytes have suggested a single [Ca2+]
i
transient followed by a steady-state increase as the characteristic [Ca2+]
i
response of these cells. In the present study, we used digital video imaging techniques in freshly dispersed myocytes from the rabbit colon, to characterize the spatiotemporal pattern of the [Ca2+]
i
signal in single cells. The distribution of [Ca2+]
i
in resting and stimulated cells was nonhomogeneous, with gradients of high [Ca2+]
i
present in the subplasmalemmal space and in one cell pole. [Ca2+]
i
gradients within these regions were not constant but showed temporal changes in the form of [Ca2+]
i
oscillations and spatial changes in the form of [Ca2+]
i
waves. [Ca2+]
i
oscillations in unstimulated cells (n = 60) were independent of extracellular [Ca2+] and had a mean frequency of 12.6 +1.1 oscillations per min. The baseline [Ca2+], was 171 ± 13 nm and the mean oscillation amplitude was 194 ± 12 nm. Generation of [Ca2+]
i
waves was also independent of influx of extracellular Ca2+. [Ca2+]
i
waves originated in one cell pole and were visualized as propagation mostly along the subplasmalemmal space or occasionally throughout the cytoplasm. The mean velocity was 23 +3 m per sec (n = 6). Increases of [Ca2+]
i
induced by different agonists were encoded into changes of baseline [Ca2+]
i
and the amplitude of oscillations, but not into their frequency. The observed spatiotemporal pattern of [Ca2+]
i
regulation may be the underlying mechanism for slow wave generation and propagation in this tissue. These findings are consistent with a [Ca2+]
i
regulation whereby cell regulators modulate the spatiotemporal pattern of intracellularly generated [Ca2+]
i
oscillations.The authors thank Debbie Anderson for excellent technical assistance with the electron microscopy and Dr. M. Regoli for providing the NK-1 agonist [Sar9,Met(O2)11]-SP. This work was supported by National Institutes of Health Grants DK 40919 and DK 40675 and Veterans Administration Grant SMI. 相似文献
20.
I-Shu Chen Chiang-Ting Chou Yuan-Yuarn Liu Chia-Cheng Yu Wei-Zhe Liang Chun-Chi Kuo 《Journal of receptor and signal transduction research》2017,37(1):1-7
Minoxidil is clinically used to prevent hair loss. However, its effect on Ca2+ homeostasis in prostate cancer cells is unclear. This study explored the effect of minoxidil on cytosolic-free Ca2+ levels ([Ca2+]i) and cell viability in PC3 human prostate cancer cells. Minoxidil at concentrations between 200 and 800?μM evoked [Ca2+]i rises in a concentration-dependent manner. This Ca2+ signal was inhibited by 60% by removal of extracellular Ca2+. Minoxidil-induced Ca2+ influx was confirmed by Mn2+-induced quench of fura-2 fluorescence. Pre-treatment with the protein kinase C (PKC) inhibitor GF109203X, PKC activator phorbol 12-myristate 13 acetate (PMA), nifedipine and SKF96365 inhibited minoxidil-induced Ca2+ signal in Ca2+ containing medium by 60%. Treatment with the endoplasmic reticulum Ca2+ pump inhibitor 2,5-ditert-butylhydroquinone (BHQ) in Ca2+-free medium abolished minoxidil-induced [Ca2+]i rises. Conversely, treatment with minoxidil abolished BHQ-induced [Ca2+]i rises. Inhibition of phospholipase C (PLC) with U73122 abolished minoxidil-evoked [Ca2+]i rises. Overnight treatment with minoxidil killed cells at concentrations of 200–600?μM in a concentration-dependent fashion. Chelation of cytosolic Ca2+ with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/AM (BAPTA/AM) did not prevent minoxidil’s cytotoxicity. Together, in PC3 cells, minoxidil induced [Ca2+]i rises that involved Ca2+ entry through PKC-regulated store-operated Ca2+ channels and PLC-dependent Ca2+ release from the endoplasmic reticulum. Minoxidil-induced cytotoxicity in a Ca2+-independent manner. 相似文献