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61.
Synaptosomal membranes were fused with liposomes using the hydration technique to produce giant proteoliposomes amenable to patch clamp recordings. Single channel currents of a cationic channel with particular properties were detected. In a solution of 150 mM NaCl, the channel displayed a unit conductance of 136 pS and a mean open state lifetime of 1.1 ms. The gating of the channel was shown to be voltage as well as calcium dependent. Pharmacological studies revealed that the channel was insensitive to a variety of channel blockers, but was inactivated by ruthenium red. Presumably, this channel may play a role in regulating the evoked release of neurotransmitters.
Offprint requests to: H. Breer 相似文献
62.
J Nittinger L Tejmar-Kolar P Fürst 《Biology of the cell / under the auspices of the European Cell Biology Organization》1990,70(3):139-142
Heat production by leukemia cells Molt 4, growing in suspensions with 1 and 3 x 10(5) cells/ml for 4 days, was studied by microcalorimetry. Heat production rates were related to cell growth, glucose consumption, lactate production and cellular ATP-content. The results show that the time course of heat dissipation is dependent on initial cell number. However, observed thermal power maxima were fairly identical in all experiments. Heat production rates per cell were similar during the initial phase of the study independently of initial cell number, while higher cell densities resulted in significantly lower rate of heat production. Glycolytic conversion of glucose into lactate is nearly stoichiometric. Our results indicate a relationship between heat production and amounts of glucose and lactate in the medium. ATP concentration in cells decreased after 24 hours of culture. 相似文献
63.
We studied the characteristics of the basal and antidiuretic hormone (arginine vasotocin, AVT)-activated whole cell currents
of an aldosterone-treated distal nephron cell line (A6) at two different cytosolic Ca2+ concentrations ([Ca2+]
c
, 2 and 30 nm). A6 cells were cultured on a permeable support filter for 10 ∼ 14 days in media with supplemental aldosterone (1 μm). At 30 nm [Ca2+]
c
, basal conductances mainly consisted of Cl− conductances, which were sensitive to 5-nitro-2-(3-phenylpropylamino)-benzoate. Reduction of [Ca2+]
c
to 2 nm abolished the basal Cl− conductance. AVT evoked Cl− conductances at 2 as well as 30 nm [Ca2+]
c
. In addition to Cl− conductances, AVT induced benzamil-insensitive nonselective cation (NSC) conductances. This action on NSC conductances was
observed at 30 nm [Ca2+]
c
but not at 2 nm [Ca2+]
c
. Thus, cytosolic Ca2+ regulates NSC and Cl− conductances in a distal nephron cell line (A6) in response to AVT. Keeping [Ca2+]
c
at an adequate level seems likely to be an important requirement for AVT regulation of ion conductances in aldosterone-treated
A6 cells.
Received: 6 May 1996/Revised: 28 June 1996 相似文献
64.
G. Menestrina C. Pederzolli M. Dalla Serra M. Bregante F. Gambale 《The Journal of membrane biology》1996,149(2):113-121
Escherichia coli hemolysin is known to cause hemolysis of red blood cells by forming hydrophilic pores in their cell membrane. Hemolysin-induced
pores have been directly visualized in model systems such as planar lipid membranes and unilamellar vesicles. However this
hemolysin, like all the members of a related family of toxins called Repeat Toxins, is a potent leukotoxin. To investigate
whether the formation of channels is involved also in its leukotoxic activity, we used patch-clamped human macrophages as
targets. Indeed, when exposed to the hemolysin, these cells developed additional pores into their membrane. Such exogenous
pores had properties very different from the endogenous channels already present in the cell membrane (primarily K+ channels), but very similar to the pores formed by the toxin in purely lipidic model membranes. Observed properties were:
large single channel conductance, cation over anion selectivity but weak discrimination among different cations, quasilinear
current-voltage characteristic and the existence of a flickering pre-open state of small conductance. The selectivity properties
of the toxin channels appearing in phospholipid vesicles were also investigated, using a specially adapted polarization/depolarization
assay, and were found to be completely consistent with that of the current fluctuations observed in excised macrophage patches.
Received: 14 August 1995/Revised: 2 October 1995 相似文献
65.
Hyperpolarization-activated K channels (K
H
channels) in the plasmalemma of guard cells operate at apoplastic pH range of 5 to over 7. Using patch clamp in a whole-cell
mode, we characterized the effect of varying the external pH between 4.4–8.1 on the activity of the K
H
channels in isolated guard cell protoplasts from Vicia faba leaves.
Acidification from pH 5.5 to 4.4 increased the macroscopic conductance of the K
H
channels by 30–150% while alkalinization from pH 5.5 to 8.1 decreased it only by roughly 15%. The voltage-independent maximum cell conductance, increased by ∼60% between pH 8.1 and 4.4 with an apparent pK
a
of 5.3, most likely owing to the increased availability of channels. Voltage-dependent gating was affected only between pH 5.5 and 4.4. Acidification in this range shifted the voltage-dependent open probability by over 10 mV. We interpret this shift as an increase of the electrical field sensed by the gating subunits
caused by the protonation of external negative surface charges. Within the framework of a surface charge model the mean spacing
of these charges was ∼30 ? and their apparent dissociation constant was 10−4.6. The overall voltage sensitivity of gating was not altered by pH changes. In a subgroup of protoplasts analyzed within the
framework of a Closed-Closed-Open model, the effect of protons on gating was limited to shifting of the voltage-dependence
of all four transition rate constants.
Received: 26 April 1996/Revised: 29 June 1996 相似文献
66.
The mechanosensitive properties of large-conductance Ca2+-activated K+ (BK) channels from embryonic rat neuroepithelium were investigated with the cell-attached and inside-out configurations of
the patch-clamp technique. The channels were activated in both recording configurations by negative pressures applied to the
patch electrode, but reversal of the effect was total and immediate in inside-out patches whereas it was incomplete and delayed
in on-cell patches. This mechanosensitivity was not mediated by Ca2+ ions or fatty acids, suggesting that it is an intrinsic property of these channels. Cytochalasin B did not affect mechanosensitivity
in on-cell patches but increased it in inside-out patches. Kinetic studies showed that stretch increased the mean open time
of the channels and decreased the slowest time constant of their closed-time distributions. The present as well as previous
results suggest complex interactions between embryonic BK channels and their membranous and submembranous environment.
Received: 1 February 1996/Revised: 25 March 1996 相似文献
67.
Zhao Rongrui Wang Wenze Wu Bowei Hoebeke Johan Hjalmarson Åke Fu Michael L. X. 《Molecular and cellular biochemistry》1996,163(1):185-193
The effects of anti-peptide antibodies against the second extracellular loop of human M2 muscarinic receptor on transmembrane potentials and currents in guinea pig single ventricular cells were analyzed using whole-cell patch clamp technique. These effects were compared with those of the muscarinic receptor agonists carbachol and acetylcholine. The antibodies shortened the action potential duration in a dose-dependent manner. By using a ramp or step rectangular pulse protocol, it was found that the antibodies increased the outward K+ current and decreased the inward basal I Ca significantly. The reversal potential of both carbachol-and antibody-induced extra currents were close to –80 mV, being in proximity to the calculated Ek of –90 mV. A -adrenergic receptor agonist, isoprenaline, prolonged the action potential and increased the overshoot which could be inhibited by both antibody and carbachol. Isoprenaline increased inward Ica and outward Ik simultaneously. Both antibody and carbachol could significantly reduce the isoprenaline-stimulated ICa but not the isoprenaline-stimulated Ik. The antibody- or carbachol-induced outward K+ current and the depressant effects of antibody and carbachol on isoprenaline-stimulated Ica were partially antagonized by atropine. These results suggest that the anti-M2 muscarinic receptor antibodies display a stimulatory activity similar to muscarinic receptor agonist on the receptor-mediated electrophysiological events. 相似文献
68.
The effects of daily supplemental chromium (200 μg) complexed with 1.8 mg nicotinic acid on plasma glucose and lipids, including
total cholesterol, HDL cholesterol, LDL cholesterol, and triglycerides, were assessed in 14 healthy adults and 5 adults with
noninsulin-dependent diabetes mellitus (NIDDM) using a double-blind crossover study with 8-wk experimental periods. Eight
of the 14 healthy subjects and all 5 subjects with NIDDM also underwent an oral glucose tolerance test with assessment of
90 min postprandial plasma glucose and insulin concentrations. No statistically significant effects of chromium nicotinic
acid supplementation were found on plasma insulin, glucose, or lipid concentrations, although chromium nicotinic acid supplementation
slightly lowered fasting plasma total and LDL cholesterol, triglycerides, and glucose concentrations, and 90-min postprandial
glucose concentrations in individuals with NIDDM. 相似文献
69.
The objective of this study was to determine whether exposure of plants to ozone (O3) increased the foliar levels of glucose, glucose sources, e.g., sucrose and starch, and glucose-6-phosphate (G6P), because in leaf cells, glucose is the precursor of the antioxidant, L-ascorbate, and glucose-6-phosphate is a source of NADPH needed to support antioxidant capacity. A further objective was to establish whether the response of increased levels of glucose, sucrose, starch and G6P in leaves could be correlated with a greater degree of plant tolerance to O3. Four commercially available Spinacia oleracea varieties were screened for tolerance or susceptibility to detrimental effects of O3 employing one 6.5 hour acute exposure to 25O nL O3 L-1 air during the light. One day after the termination of ozonation (29 d post emergence), leaves of the plants were monitored both for damage and for gas exchange characteristics. Cultivar Winter Bloomsdale (cv Winter) leaves were least damaged on a quantitative grading scale. The leaves of cv Nordic, the most susceptible, were approximately 2.5 times more damaged. Photosynthesis (Pn) rates in the ozonated mature leaves of cv Winter were 48.9% less, and in cv Nordic, 66.2% less than in comparable leaves of their non-ozonated controls. Stomatal conductance of leaves of ozonated plants was found not to be a factor in the lower Pn rates in the ozonated plants. At some time points in the light, leaves of ozonated cv Winter plants had significantly higher levels of glucose, sucrose, starch, G6P, G1P, pyruvate and malate than did leaves of ozonated cv Nordic plants. It was concluded that leaves of cv Winter displayed a higher tolerance to ozone mediated stress than those of cv Nordic, in part because they had higher levels of glucose and G6P that could be mobilized during diminished photosynthesis to generate antioxidants (e.g., ascorbate) and reductants (e.g., NADPH). Elevated levels of both pyruvate and malate in the leaves of ozonated cv Winter suggested an increased availability of respiratory substrates to support higher respiratory capacity needed for repair, growth, and maintenance.Abbreviations ADPG-PPiase
ADPglucose pyrophosphorylase
- ASC
L-ascorbic acid
- APX
ascorbate peroxidase
- Ce
CO2 concentration in air in the measuring cuvette during photosynthesis measurements
- Ci
CO2 concentration in the leaf intercellular spaces during photosynthesis measurement
- Chl
chlorophyll
- DHA
dehydroascorbic acid
- DHA reductase
dehydroascorbate reductase
- DHAP
dihydroxyacetone phosphate
- GAP
glyceraldehyde-3-phosphate
- Gluc
glucose
- GR
glutathione reductase
- Gsw
stomatal conductance with units as mmol H2O m-2 s-1
- GSSG
oxidized glutathione
- GSH
reduced glutathione
- G1P
glucose-1-phosphate
- G6P
glucose-6-phosphate
- G6P dehydrogenase
glucose-6-phosphate dehydrogenase
- 6PG
6-phosphogluconate
- 6PG dehydrogenase
6-phosphogluconate dehydrogenase
- F6P
fructose-6-phosphate
- FBP
fructose-1,6-bisphosphate
- MAL
malate
- MDHA reductase
monodehydroascorbate reductase
- PE
post-emergence
- PEP
phosphoenolpyruvate
- PGA
3-phosphoglycerate
- Pi
orthophosphate
- PYR
pyruvate
- Pn
net CO2 photoas-similation in leaves
- PPFD
photosynthetic photon flux density with units of mol photons m-2 s-1
- PPRC
pentose phosphate reductive cycle
- RuBP
ribulose-1,5-bisphosphate
- rubisco
ribulose-1,5-bisphosphate carboxylase/oxygenase
- SLW
specific leaf weight
- TCA cycle
tricarboxylic acid cycle
- Triose-P
DHAP+GAP 相似文献
70.
The heterogeneous nature of microbial products as shown by solid-state13C CP/MAS NMR spectroscopy 总被引:1,自引:1,他引:0
Homoionic Na-, Ca-, and Al-clays were prepared from the <2 m fractions of Georgia kaolinite and Wyoming bentonite and mixed with sand to give artificial soils with 5, and 25% clay. The artificial soils were inoculated with microbes from a natural soil before incubation. Unlabelled and uniformly13C-labelled (99.9% atom) glucose were incorporated into the artificial soils to study the effects of clay types, exchangeable cations and clay contents on the mineralization of glucose-carbon and glucose-derived organic materials. Chemical transformation of glucose-carbon upon incorporation into microbial products and metabolites, was followed using solid-state13C CP/MAS NMR spectroscopy.There was a significant influence of exchangeable cations on the mineralization of glucose-carbon over a period of 33 days. At 25% clay content, mineralization of glucose-carbon was highest in Ca-soils and lowest in Al-soils. The influence of exchangeable cations on mineralization of glucose-carbon was more pronounced in soils with bentonite clay than those with kaolinite clay. Statistical analysis of data showed no overall effect of clay type on mineralization of glucose-carbon. However, the interactions of clay type with clay content and clay type with clay content and exchangeable cations were highly significant. At 25% clay content, the mineralization of glucose-carbon was significantly lower in Na- and Al-soils with Wyoming bentonite compared with Na- and Al-soils with Georgia kaolinite. For Ca-soils this difference was not significant. Due to the increased osmotic tension induced by the added glucose, mineralization of glucose-carbon was slower in soils with 5% clay than soils with 25% clay.Despite the differences in the chemical and physical characteristics of soils with Ca-, Na- and Al-clays, the chemical composition of organic materials synthesised in these soils were similar in nature. Assuming CP/MAS is quantitative, incorporation of uniformly13C-labelled glucose (99.9% atom) in these soils resulted in distribution of carbon in alkyl (24–25%), O-alkyl (56–63%), carbonyl (11–15%) and small amounts of aromatic and olefinic carbon (2–4%). However, as decomposition proceeded, the chemistry of synthesised material showed some changes with time. In the Ca- and Na-soils, the proportions of alkyl and carbonyl carbon decreased and that of O-alkyl carbon increased with time of incubation. However, the opposite trend was found for the Al-soil.Proton-spin relaxation editing (PSRE) subspectra clearly showed heterogeneity within the microbial products. Subspectra of the slowly-relaxing (long T1(H)) domains were dominated by alkyl carbon in long- and short-chain structures. The signals due to N-alkyl (55 ppm) and carbonyl carbon were also strong in these subspectra. These subspectra were very similar to those obtained for microbial and fungal materials and were probably microbial tissues attached to clay surfaces by polysaccharide extracellular mucilage. Subspectra of fast-relaxing (short T1(H)) domains comprised mostly O-alkyl and carbonyl carbon and were probably microbial metabolites released as neutral and acidic sugars into the extracellular environment, and strongly sorbed by clay surfaces. 相似文献