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1.
The patch-clamp technique was applied to vacuoles isolated from a photoautotrophic suspension cell culture of Chenopodium rubrum L. and vacuolar clamp currents, which are predominantly carried by the previously identified Ca2+-dependent slow vacuolar (SV) ion channels, were recorded. These currents, which were activated by 1-s voltage pulses of -100 mV (vacuolar interior negative) in the presence of 100 M Ca2+ (cytosolic side), could be blocked completely and reversibly by the calmodulin antagonist W-7 [N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide] and its chlorine-deficient analogue W-5; half-maximum inhibition was found at approx. 6 M for W-7 and 70 M for W-5. Inhibition was reversed by addition of 1 g · ml–1 calmodulin purified from Chenopodium cell suspensions; reversal by bovine brain calmodulin was scarcely appreciable. We conclude that cytosolic calmodulin mediates the Ca2+ dependence of the SV-channel in the Chenopodium tonoplast.Abbreviations SV-channel
slowly activated, vacuolar ion channel
- W-5
N-(6-aminohexyl)-1-naphthalenesulfonamide
- W-7
N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide
We acknowledge support by the Deutsche Forschungsgemeinschaft and the Bundesminister für Forschung und Technologie, Bonn, and by the Justus-Liebig-Universität Giessen (to W.B.) 相似文献
2.
A soluble protein was isolated from Mougeotia by chloropromazine-sepharose 4 B affinity chromatography. The protein matches the properties of calmodulin in terms of heat stability, Ca2+-dependent electrophoretic mobility in sodium-dodecyl-sulfate polyacrylamide gels, and its ability to activate cyclic nucleotide phosphodiesterase in a Ca2+-dependent manner. Phytochrome-mediated chloroplast reorientational movement in Mougeotia was inhibited by the calmodulin antagonist trifluoperazine, a hydrophobic compound, or N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7), a hydrophilic compound; 50% inhibition (IC50) of chloroplast movement is caused by 20–50 mol l-1 trifluoperazine or 100 mol l-1 W-7. The Ca2+-calmodulin may act as an intermediate in the chloroplast reorientational response in Mougeotia governed by phytochrome.Abbreviations EGTA
ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid
- SDS
sodium dodecyl sulfate
- W-7
N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide 相似文献
3.
Crude Ca2+-activated protein kinase from membranes of apple (Malus domestica L. Borkh., Cox's Orange Pippin) fruit can be partially purified to yield a Ca2+-dependent protein kinase whose activity is apparently not regulated by calmodulin. The autophosphorylating catalytic subunit of this protein kinase shows a Ca2+-dependent mobility shift of approx. 10 kilodaltons (kDa) on sodium dodecyl sulphate-polyacrylamide gel electrophoresis; in the absence of added Ca2+ or ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid (EGTA) its apparent molecular mass is approx. 50 kDa. The Ca2+-dependent protein kinase is inhibited by the calmodulin antagonists N-(6-aminohexyl)-5-chloro-1-naphthalenesulphonamide and trifluoperazine with IC50 values of approx. 45 M and 15 M, respectively. These similarities between the protein kinase and calmodulin indicate that the kinase may be a calmodulin-like protein.Abbreviations DEAE
diethylaminoethyl
- EGTA
ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid
- Hepes
4-(-2-hydroxyethyl)-1-piperazineethanesulphonic acid
- kDa
kilodalton
- PAGE
polyacrylamide gel electrophoresis
- SDS
sodium dodecyl sulphate
- W7
N-(6-aminohexyl)-5-chloro-1-naphthalenesulphonamide
- W5
N-(6-aminohexyl)-naphthalenesulphonamide 相似文献
4.
Ichida S Abe J Zhang YA Sugihara K Imoto K Wada T Fujita N Sohma H 《Neurochemical research》2000,25(12):1629-1635
The characteristics of the inhibitory effect of calcium ion (Ca2+)/calmodulin (CaM) on specific [125I]-omega-conotoxin GVIA (125I--CTX) binding and on the labeling of 125I--CTX to crude membranes from chick brain were investigated. The inhibitory effect of Ca2+/CaM depended on the concentrations of free Ca2+ and CaM. The IC50 values for free Ca2+ and CaM were about 2.0 × 10–8 M and 3.0 g protein/ml, respectively. The inhibitory effect of Ca2+/CaM was attenuated by the CaM antagonists W-7, prenylamine and CaM-kinase II fragment (290–309), but not by the calcineurin inhibitor FK506. Ca2+/CaM also inhibited the labeling of a 135-kDa band (which was considered to be part of N-type Ca2+ channel 1 subunits) with 125I--CTX using a cross-linker. These results suggest that Ca2+/CaM affects specific 125I--CTX binding sites, probably N-type Ca2+ channel 1 subunits, in crude membranes from chick whole brain. 相似文献
5.
Callus was initiated from immature leaf and stem segments of rose (Rosa hybrida cv. Landora) and subcultured every four weeks on a basal medium of half-strength Murashige & Skoog (1962) salts plus 30 g l-1 sucrose (1/2 MS) and supplemented with 2.2 M BA, 5.4 M NAA and 2.2–9.0 M 2,4-D. Embryogenic callus and subsequently somatic embryos were obtained from 8-week-old callus culture on 1/2 MS+2.2 M BA+0.05 M NAA+0.3 M GA3+200–800 mg l-1 L-proline. Long-term cultures were established and maintained for up to 16 months by repeated subculture of embryogenic callus on L-proline deficient medium. About 12% of cotyledonary stage embryos taken from cultures cold-stored at 8±1°C for 4 days germinated on 1/2 MS+2.2 M BA+0.3 M GA3+24.7 M adenine sulphate.Abbreviations BA
benzyladenine
- NAA
-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid 相似文献
6.
Summary The actin-activated ATPase activityPhysarum myosin was shown to be inhibited of M levels of Ca2+. To determine if Ca2+ regulates ATP-dependent movement ofPhysarum myosin on actin, latex beads coated withPhysarum myosin were introduced intoChara cells by intracellular perfusion. In perfusion solution containing EGTA, the beads moved along the parallel arrays ofChara actin filaments at a rate of 1.0–1.8 m/sec; however, in perfusion solution containing Ca2+, the rate reduced to 0.0–0.7 m/sec. The movement of beads coated with scallop myosin, whose actin-activated ATPase activity is activated by Ca2+, was observed only in the perfusion solution containing Ca2+, indicating that myosin is responsible for the inhibitory effect of Ca2+ onPhysarum myosin movement. The involvement of this myosin-linked regulation in the inhibitory effect of Ca2+ on the cytoplasmic streaming observed inChara internodal cell andPhysarum plasmodium was discussed.Abbreviations ATP
adenosine 5-triphosphate
- DTT
dithiothreitol
- EDTA
ethylenediaminetetraacetic acid
- EGTA
ethyleneglycolbis(-aminoethylether) N,N,N,N-tetraacetic acid
- PIPES
piperazine-N,N-bis(2-ethanesulfonic acid) 相似文献
7.
Jean-Michel Wieruszeski Jean-Claude Michalski Jean Montreuil Gérard Strecker 《Glycoconjugate journal》1990,7(1):13-26
The major pentasaccharides Fuc(1-2)[GalNAc(1-3)]Gal(1-4)[Fuc(1-3)]Glc and Fuc(1-2) [Gal(1-3)]Gal(1-4)[Fuc(1-3)]Glc, which are normally present in the urine of bloodgroup A Leb and B Leb healthy subjects, were each found to be contaminated by a minor component when analysed by1H-NMR. The determination of these structures, Fuc(1-2) [GalNAc(1-3)]Gal(1-3)[Fuc(1-4)]Glc and Fuc(1-2) [Gal(1-3)]Gal(1-3)[Fuc(1-4)]Glc, was based on the results of methylation analysis and1H/13C-NMR spectroscopy.Abbreviations HPLC
high performance liquid chromatography
- GLC
gas liquid chromatography
- NMR
nuclear magnetic resonance
- COSY
correlation spectroscopy
- Gal
d-galactopyranose
- GalNAc
2-acetamido-2-deoxy-d-galactopyranose
- Glc
d-glucopyranose
- Fuc
l-fucopyranose
- LNDFH I
lacto-N-difucohexaose I (Leb determinant 相似文献
8.
Melanie S. Johnson James Simpson David J. MacEwan Angela Ison Roger A. Clegg Kevin Connor Rory Mitchell 《Molecular and cellular biochemistry》1995,146(2):127-137
The characteristics of PKC activation induced by a number of compounds were investigated using PKCs, partially-purified from sources with a naturally high abundance of certain Ca2+ dependent PKC isoforms. Native isoforms were used rather than PKC isoforms expressed from a baculovirus system to assess the effect of tissue specific factors on activity. However, some data using recombinant PKC were included for comparison.The presence of specific PKC isoforms in different tissues was determined using Western blot analysis. Protein kinase C , 1, , , and / were all present in rat midbrain cytosolic extract, PKC , 1, , and / were present in spleen cytosol, and PKC and / were present in COS 7 cell cytosol. The predominance of and activities in COS 7 and spleen extracts respectively was confirmed by enzymic assay.The PKC activity assay was configured such that the Ca2+ dependence of the PKC activity induced by different PKC activators could be determined. Phorbol 12,13-dibutyrate (PDBu) was virtually equipotent on the Ca2+-dependent PKC activity from midbrain and spleen and slightly less potent on that from COS 7 cells. In the absence of Ca2+, PDBu was considerably less potent overall (as, indeed, were the other PKC activators) and was less potent on COS 7 cell PKC than on those from midbrain or spleen. Mezerein was more potent than PDBu at inducing PKC activity in COS 7 cell extracts in either the absence or presence of Ca2+ whereas in the presence of Ca2+, mezerein was slightly less potent on midbrain and spleen than PDBu and equipotent in the absence of Ca2+. Maximum values for Ca2+-independent activation by mezerein indicated that this activator was particularly effective in recruiting Ca2+-dependent PKC isoform activity in a Ca2+ free environment. The greater potency of mezerein on PKC was confirmed using PKC and further purified from rat spleen by hydroxylapatite (HAP) chromatography. The effects of both PDBu and mezerein were investigated using anterior pituitary tissue where a particularly high potency of mezerein in the absence of Ca2+ was noted. The diacylglycerol, 1,2-dioctanoyl-sn-glycerol (DOG), appeared to cause little or no activation of native Ca2+-dependent isoforms in Ca2+ free conditions unlike another longer chain diacylglycerol, 1,2-dioleoyl-sn-glycerol. Also DOG activated midbrain PKCs more potently than PKCs from spleen or COS 7 cells (or lung and pituitary tissue) in the presence of Ca2+. The concentration-dependence of DOG was examined on PKC and PKC further purified from brain by HAP chromatography, revealing that DOG was equally potent on both of these isoforms derived from brain and on recombinant PKC . However, [3H]PDBu binding data using PKC purified from several sources gave very different IC50 values when DOG was used as a displacer, and in general these values correlated with the EC50 values recorded from the activity assay.The data presented here indicate that there are distinct differences in the activator pharmacology of different native PKC isoforms and between the same isoform expressed in different tissues, either because of post-translational modifications or some other tissue specific factor. 相似文献
9.
Gerard Strecker Jean-Michel Wieruszeski Jean-Claude Michalski Jean Montreuil 《Glycoconjugate journal》1988,5(4):385-396
The structure of a new nonasaccharide isolated from human milk has been investigated. By using methylation analysis, FAB-MS and1H-and13C-NMR spectroscopy as basic methods of structural investigation, this oligosaccharide was identified as VI2--Fuc,V4-Fuc,III3--Fuc-p-lacto-n-hexaose: Fuc1-2Gal1-3[Fuc1-4]GlcNAc1-3Gal1-4[Fuc1-3]GlcNAc1-3Gal1-4Glc.Abbreviations COSY
correlation spectroscope
- DP
degree of polymerisation
- FAB-MS
fast atom bombardment-mass spectrometry
- HPLC
high performance liquid chromatography
- NMR
nuclear magnetic resonance
- GLC
gas-liquid chromatography 相似文献
10.
Summary In internodal cells ofLamprothamnium succinctum, turgor regulation in response to hypotonie treatment is inhibited by lowering external Ca2+ concentration ([Ca2+]e) from 3.9 (normal) to 0.01 (low) mM. In order to clarify whether a change in the cytoplasmic free Ca2+ concentration ([Ca2+]c) is involved in turgor regulation, the Ca2+ sensitive protein aequorin was injected into the cytoplasm of internodal cells. A large transient light emission was observed upon hypotonic treatment under normal [Ca2+]e but not under low [Ca2+]e. Thus hypotonic treatment induces a transient increase in [Ca2+]c under normal [Ca2+]e but not under low [Ca2+]e.Abbreviations ASW
artificial sea water
- i
cellular osmotic pressure
- [Ca2+]c
cytoplasmic free Ca2+ concentration
- EDTA
ethylenediamine-tetraacetic acid
- EGTA
ethylenglycol-bis(-aminoethyl ether(N,N-tetraacetic acid
- [Ca2+]e
external Ca2+ concentration
- e
external osmotic pressure
- GM
glass micropipette
- GP
glass plate
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethansulfonic acid
- MS
microscope stage
- OL
objective lens
- PIPES
piperazine-N-N-bis(2-ethanesulfonic acid)
- W
Weight 相似文献
11.
Summary Treatment of Allium cepa L. cellsuspension cultures with a biotic elicitor derived from the fungus Botrytis cinerea, resulted in phytoalexin synthesis. Two phytoalexins, 5-octylcyclopenta-1,3-dione and 5-hexyl-cyclopenta-1,3-dione, were accumulated in cultured onion cells. Removal of extracellular Ca2+ by the calcium chelator ethylene glycol bis(b-aminoethyl ether) N,N-tetraacetic acid abolished the elicitor-mediated phytoalexin synthesis. The calcium channel blockers, verapamil and 8-N,N-(dimethylamino)octyl-3,4,5-trimethoxybenzoate caused similar effects, whereas the addition of the Ca2+ ionophore A23187 enhanced the accumulation of phytoalexins in the absence of the elicitor. Increase in the cytoplasmic Ca2+ concentration in elicitor-treated onion cells was observed as monitored by the fluorescent calcium indicator indo-1. These observations suggest that Ca2+ acts as a second messenger in the regulation of phytoalexin synthesis in cultured onion cells.Abbreviations A23187
4-bromo-calcium ionophore
- cAMP
adenosine 3,5-cyclic monophosphate
- [Ca2+]cyt
cytoplasmic Ca2+ concentration
- EGTA
ethylene glycol bis(b-aminoethyl ether) N,N-tetraacetic acid
- EtOH
ethanol
- Et2O
diethyl ether
- fr.wt
fresh weight
- HR
hypersensitive response
- PIPES
piperazine N,N-bis-(2-ethanesulfonic acid)
- TMB-8
[8-N,N-(dimethylamino)] octyl-3,4,5-trimethoxy-benzoate
- Tsl
tsibulin 相似文献
12.
Summary We examined the ionic regulation of tip growth inNeurospora crassa by a combination of electrophysiology and confocal microscopy. To determine if transmembrane ionic fluxes are required for tip growth, we voltage clamped the membrane from –200 to +50 mV. In this voltage range, transmembrane ionic fluxes would either reverse (e.g., K+) or change dramatically (e.g., Ca2+ influx) but had no effect on hyphal growth rates. Therefore, ionic fluxes (including Ca2+ influx) may not be required for tip growth. However, intracellular Ca2+ may still play an obligatory role in tip growth. To assess this possibility, we first increased cytosolic Ca2+ directly by ionophoresis. Elevated Ca2+ induced subapical branch initiation, often multiple tips. At hyphal tips, fluorescence ratio imaging using fluo-3 and fura-red revealed a pronounced tip-high Ca2+ gradient within 10 m of the tip in growing hyphae which was not observed in nongrowing hyphae. Injection of the Ca2+ chelator 1,2-bis(ortho-aminophenoxy)ethane-N,N,N,N-tetrapotassium acetate consistently inhibited growth concomitantly with a depletion of intracellular Ca2+ and dissipation of the tip-high gradient. We conclude that Ca2+ plays a regulatory role in tip initiation and the maintenance of tip growth. Because plasma membrane ionic fluxes do not play a role in tip growth, we suggest that the tip-high Ca2+ gradient is generated from intracellular Ca2+ stores in the ascomyceteN. crassa.Abbreviations BAPTA
1,2-bis(ortho-aminophenoxy)ethane-N,N,N,N-tetrapotassium acetate
- [Ca2+]i
intracellular Ca2+ concentration
- fluo-3
2,7-dichloro-6-hydroxy-3-oxo-9-xanthenyl-4-methyl-2,2-(ethylenedioxy)dianiline-N,N,N,N-tetraacetic acid 相似文献
13.
The Ca2+-binding properties of calmodulin purified from zucchini (Cucurbita pepo L.) has been determined. A value of 3.3 mol Ca2+ per mol of zucchini calmodulin was measured at pH 7.5 by equilibrium chromatography. The far-and near-UV circular-dichroic spectra of the Ca2+-and Mg2+-saturated as well as from the metal-free forms of zucchini calmodulin reveal that upon Ca2+-binding the -helix content increases. A comparison with the spectra of vertebrate calmodulin indicates that both calmodulin have a similar secondary structure, similar Ca2+-induced conformational changes and the same number of Ca2+-binding sites.Abbreviations CAPP
10-(3-aminopropyl)-2-chloro-phenothiazine
- EGTA
ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid
- EDTA
ethylenediaminetetraacetic acid
Dedicated to Prof. Dr. Karl Decker on the occasion of his 60th birthday 相似文献
14.
Summary To control the intracellular free Ca2+ concentration from the cell exterior, pollen tubes ofLilium longiflorum were treated with a Ca2+ ionophore, A23187. Cytoplasmic streaming was inhibited when the free Ca2+ concentration of the external medium ([Ca2+]) was raised to 5×10–6 M or higher. At [Ca2+] below 1×10–6 M, the rhodamine-phalloidin stained actin filaments appeared straight and thin. However, at [Ca2+] which inhibited cytoplasmic streaming, the actin filaments appeared fragmented. In pollen tubes, Ca2+ regulation of cytoplasmic streaming may be linked not only to myosin (Shimmen 1987) but also to actin.Abbreviations ATP
adenosine-5-triphosphoric acid
- [Ca2+]
concentration of free Ca2+
- EGTA
ethyleneglycol-bis-(-aminoethylether)N,N,N,N-tetraacetic acid
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid
- PIPES
piperazine-N,N-bis(2-ethanesulfonic acid)
- Rh-ph
rhodamine-conjugated phalloidin 相似文献
15.
Anton F. Post 《Archives of microbiology》1986,145(4):353-357
Transitions in growth irradiance level from 92 to 7 Em-2 s-1 and vice versa caused changes in the pigment contents and photosynthesis of Oscillatoria agardhii. The changes in chlorophyll a and C-phycocyanin contents during the transition from high to low irradiance (HL) were reflected in photosynthetic parameters. In the LH transition light utilization efficiencies of the cells changed faster than pigment contents. This appeared to be related to the lowering of light utilization efficiencies of photosynthesis. As a possible explanation it was hypothesized that excess photosynthate production led to feed back inhibition of photosynthesis. Time-scales of changes in the maximal rate of O2 evolution were discussed as changes in the number of reaction centers of photosystem II in relation to photosynthetic electron transport. Parameters that were subject to change during irradiance transitions obeyed first order kinetics, but hysteresis occurred when comparing HL with LH transients. Interpretation of first order kinetic analysis was discussed in terms of adaptive response vs changes in growth rate.Non-standard abbreviations Chla
chlorophyll a
- CPC
C-phycocyanin
- PS II
photosystem II
- PS I
photosystem I
- RC II
reaction center of photosystem II
- P
photosynthetic O2-evolution
- I
irradiance, Em-2 s-1
-
light utilization efficiency of cells, mmol O2·mg dry wt-1·h-1/Em-2 s-1
-
light utilization efficiency of photosynthetic apparatus, mol O2·mol Chla
-1·h-1/Em-2 s-1
- Pmax
maximal rate of O2 evolution by cells, mol O2·mg dry wt-1·h-1
- Pmax
maximal rate of O2 evolution by photosynthetic apparatus, mol O2·mol·Chla
-1·h-1
- LL
low light, E m-2 s-1
- HL
high light, E m-2 s-1
- LH
low to high light transition
- HL
high to low light transition
-
k
specific rate of adaptation, h-1
-
specific growth rate, h-1
-
Q
pool size of cell constituent, mol·mg dry wt-1
-
q
net synthesis rate of cell constituent, mol·mg dry wt-1·h-1 相似文献
16.
Effect of transmembrane Ca2+ gradient on the coupling of β-adrenergic receptors and adenylyl cyclase
In order to investigate the effect of transmembrane Ca2+ gradient on Gs mediated coupling of -AR and adenylyl cyclase, -AR from duck erythrocytes and Gs and adenylyl cyclase from bovine brain cortices were co-reconstituted into asolectin liposomes with different transmembrane Ca2+ gradient. These proteoliposomes were proven to be impermeable to water-soluble substances. The results obtained indicate that a physiological transmembrane Ca2– gradient (1000-fold) is essential for higher stimulation of adenylyl cyclase by hormone-activated -AR via coupling to Gs and can be further enhanced by the decrease of such Ca2+ gradient within certain range (100 fold) following Ca2+ influx into cells during signal transduction. Fluorescence polarization of DPH revealed that transmembrane Ca2+ gradient modulates adenylyl cyclase and its stimulation by hormones through mediating a change in lipid fluidity. Correspondent conformational changes of -AR were also detected from the fluorescence spectra and quenching of Acrylodan-labelled -AR in those proteoliposomes. It is suggested that a proper transmembrane Ca2+ gradient is essential for the optimal fluidity of the phospholipid bilayer in the proteoliposomes, which favors the formation of a suitable conformation of the reconstituted -AR and thus promotes the stimulation of adenylyl cyclase activities by hormone-activated -AR via Gs.Abbreviations ATP
adenosine triphosphate
- -AR
-adrenergic receptors
- AC
adenylyl cyclase
- DHA
dihydroalprenolol
- DPH
diphenylhexatriene
- [Ca2+]i
Ca2+ concentration inside proteoliposomes
- [Ca2+]o
Ca2+ concentration outside proteoliposomes
- cAMP
cyclic adenosine monophosphate
- DTT
Dithiothreitol
- FS
fluorescein sulfonate
- Gs
Stimulatory GTP-binding protein
- GTP
guanosine triphosphate
- GTPS
guanosine 5-O-(3-thiotriphosphate)
- kDa
kilodalton
- SDS
sodium dodecyl sulfate
- Tris
N-tris(hydroxymethyl)aminomethane 相似文献
17.
The Ca2+ channel 1B subunit is a pore-forming component capable of generating N-type Ca2+ channel activity. Although N-type Ca2+ channel plays a role in a variety of neuronal functions, 1B-deficient mice exhibit normal life span without apparent abnormalities of behavior, histology or plasma norepinephrine level, presumably owing to compensation by some other Ca2+ channel 1 or subunit. In this study, we studied the levels of 1A, 1C, 1D, 1E, 1, 2, 3 and 4 mRNAs in adrenal gland of 1B-deficient mice. The 1A mRNA in homozygous mice was expressed at higher level than in wild or heterozygous mice, but no difference in the expression levels of 1C, 1D, 1E, 1, 2, 3 and 4 was found among wild, heterozygous and homozygous mice. The protein level of 1A in homozygous mice was also expressed at higher level than in wild or heterozygous mice. To examine whether increased expression is induced by cis-regulatory element within 5-upstream region of 1A gene, we examined lacZ expression in 1B-deficient × 1A6.3-lacZ mice (carrying a 6.3-kb 5-upstream fragment of 1A gene fused to E. coli lacZ reporter gene), which express lacZ in medullar chromaffin cells, but not in cortex. The levels of lacZ expression in homozygous 1B-deficient × 1A6.3-lacZ mice were higher than in wild or heterozygous mice. Therefore, a possible explanation of the normal behavior and plasma norepinephrine level of 1B-deficient mice is that compensation by 1A subunit occurs and that 6.3-kb 5-upstream region of 1A gene contains enhancer cis-element(s) for compensation in adrenal medulla chromaffin cells. (Mol Cell Biochem 271: 91–99, 2005) 相似文献
18.
Evidence for two modes of Ca2+ entry following muscarinic stimulation of a human salivary epithelial cell line 总被引:1,自引:0,他引:1
Xinjun He Xiaozai Wu R. James Turner Bruce J. Baum 《The Journal of membrane biology》1990,115(2):159-166
Summary We have investigated muscarinic receptor-operated Ca2+ mobilization in a salivary epithelial cell line, HSG-PA, using an experimental approach which allows independent evaluation of intracellular Ca2+ release and extracellular Ca2+ entry. The carbachol (Cch) dose response of intracellular Ca2+ release indicates the involvement of a single, relatively low-affinity, muscarinic receptor site (K
0.510 or 30 m, depending on the method for [Ca2+]
i
determination). However, similar data for Ca2+ entry indicate the involvement of two Cch sites, one consistent with that associated with Ca2+ release and a second higher affinity site withK
0.52.5 m. In addition, the Ca2+ entry response observed at lower concentrations of Cch (2.5 m) was completely inhibited by membrane depolarization induced with high K+ (>55mm) or gramicidin D (1 m), while membrane depolarization had little or no effect on Ca2+ entry induced by 100 m Cch. Another muscarinic agonist, oxotremorine-M (100 m; Oxo-M), like Cch, also induced an increase in the [Ca2+]
i
of HSG-PA cells (from 72±2 to 104±5nm). This response was profoundly blocked (75%) by the inorganic Ca2+ channel blocker La3+ (25–50 m) suggesting that Oxo-M primarily mobilizes Ca2+ in these cells by increasing Ca2+ entry. Organic Ca2+ channel blockers (verapamil or diltiazem at 10 m, nifedipine at 1 m), had no effect on this response. The Oxo-M induced Ca2+ mobilization response, like that observed at lower doses of Cch, was markedly inhibited (70–90%) by membrane depolarization (high K+ or gramicidin D). At 100 m Cch the formation of inositol trisphosphate (IP3) was increased 55% above basal levels. A low concentration of carbachol (1 m) elicited a smaller change in IP3 formation (25%), similar to that seen with 100 m Oxo-M (20%). Taken together, these results suggest that there are two modes of muscarinic receptor-induced Ca2+ entry in HSG-PA cells. One is associated with IP3 formation and intracellular Ca2+ release and is independent of membrane potential; the other is less dependent on IP3 formation and intracellular Ca2+ release and is modulated by membrane potential. This latter pathway may exhibit voltage-dependent gating. 相似文献
19.
Phytochrome is confirmed to be the photoreceptor pigment in the germination response of Onoclea sensibilis L. by demonstrating red-far-red (R-FR) photoreversibility. External Ca2+ is required for this response with a threshold at a submicromolar concentration. Ethylene glycol-bis(-amino-ethyl ether)-N,N,N,N-tetraacetic acid, La3+ and Co2+ reversibly inhibit germination. Lanthanum only inhibits germination when applied before or during irradiation, indicating that the external Ca2+ requirement is transient, although in the absence of Ca2+ the R-stimulated system remains maximally poised to accept the ion for over 4 h after irradiation. The ability to respond to Ca2+ 4.1 h after R-irradiation is not reversed by FR-irradiation, indicating that Ca2+ transport has been uncoupled from phytochrome. Barium and Sr2+, but not Mg2+ can substitute for Ca2+. Artificially increasing the concentration of intracellular free Ca2+ with the ionophore A 23187 stimulates germination in the dark. The Ca2+-calmodulin antagonists, trifluoperizine and chlorpromazine, reversibly inhibit germination. Calcium is required in phytochrome-mediated fern spore germination; it may be acting as a second messenger.Abbreviations EGTA
ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid
- FR
far-red light
- R
fed light 相似文献
20.
V. Varga R. Janáky K-M Marnela Pirjo Saransaari S. S. Oja 《Neurochemical research》1994,19(3):243-248
The in vitro effects of -L-glutamyltaurine on different stages of excitatory aminoacidergic neurotransmission were tested with -D-glutamyltaurine as reference. -L-Glutamyltaurine enhanced the K+-stimulated release of [3H]glutamate from cerebral cortical slices (25% at 0.1 mM) and slightly inhibited the uptake by crude brain synaptosomal preparations (about 10% at 1 mM). -L-Glutamyltaurine was also a weak displacer of glutamate and its agonists from their binding sites in brain synaptic membrane preparations, being, however, less selective to quisqualate (QA) sites than -D-glutamyltaurine. The basal influx of Ca2+ into cultured cerebellar granular cells was not affected by 1 mM -L-glutamyltaurine, but the glutamate- and its agonist-activated influx was significantly inhibited in low-Mg2+ (0.1 mM) and Mg2+-free media. The glutamate-evoked increase in free intracellular Ca2+ and the kainate-activated formation of cGMP in cerebellar slices were both markedly inhibited by 0.1 mM -L-giutamyltaurine. We propose that -L-glutamyltaurine may act as endogenous modulator in excitatory aminoacidergic neurotransmission. 相似文献