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201.
The effect of mechanical stress on the heart's electrical activity has been termed mechanoelectric feedback. The response to stretch depends upon the magnitude and the waveform of the stimulus, and upon the timing relative to the cardiac cycle. Stretch-activated ion channels (SACs) have been regarded as the most likely candidates for serving as the primary transducers of mechanical stress. We explored the steady state and dynamic responses of single channels in adult rat atrial cells using the patch clamp with a pressure clamp. Surprisingly, we only observed K+-selective SACs, probably of the 2P domain family. The channels were weakly outward rectifying with flickery bursts. In cell attached mode, the mean conductance was 74±14 and 65±16 pS for +60 and −60 mV, respectively (140 mM [K+]out, 2 mM [Mg2+]out and 0 mM [Ca2+]out). The latency of the response to pressure steps was 50–100 ms and the time to peak 400 ms. About half of the channels in cell-attached patches showed adaptation/inactivation where channel activity declined to a plateau of 20–30% of peak in 1 s. The time dependent behavior of these SACs is generally consistent with whole-cell currents observed in chick and rat ventricular cells, although the net current was outward rather than inward. 相似文献
202.
Intracellular inositol 1,4,5-trisphosphate receptors (IP(3)Rs) form tetrameric Ca2+-release channels that are crucial for Ca2+ signalling in many eukaryotic cells. IP(3)R subunits contain an N-terminal, cytoplasmic, ligand binding domain linked by a modulatory domain to a channel-forming, hydrophobic C-terminal domain. We assembled and sequenced cDNAs encoding the SI-/SII+/SIII+ splice variant of the human brain type I IP(3)R, and functionally expressed the full-length receptor, and a C-terminally truncated receptor lacking the final 20% of the protein, in mammalian and insect cells. Both proteins were insoluble, consistent with in vivo immunofluorescence and ligand binding studies. This contrasted with the behaviour of recombinant FIKBP12 (a soluble control protein). The truncated receptor also fractionated with the "membrane" pellet after alkaline carbonate treatment. We conclude that the human type I IP(3)R forms high MW aggregates or complexes in cells when expressed without the C-terminal hydrophobic domain. This behaviour should be considered when expressing and refolding "soluble" human type I IP(3)R domains for structural studies. 相似文献
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204.
Membrane permeability of fructose-1,6-diphosphate in lipid vesicles and endothelial cells 总被引:3,自引:0,他引:3
Ehringer WD Niu W Chiang B Wang OL Gordon L Chien S 《Molecular and cellular biochemistry》2000,210(1-2):35-45
Fructose-1,6-diphosphate (FDP) is a glycolytic intermediate which has been used an intervention in various ischemic conditions for two decades. Yet whether FDP can enter the cell is under constant debate. In this study we examined membrane permeability of FDP in artificial membrane bilayers and in endothelial cells. To examine passive diffusion of FDP through the membrane bilayer, L-a-phosphatidylcholine from egg yolk (Egg PC) (10 mM) multi-lamellar vesicles were created containing different external concentrations of FDP (0, 0.5, 5 and 50 mM). The passive diffusion of FDP into the vesicles was followed spectrophotometrically. The results indicate that FDP diffuses through the membrane bilayer in a dose-dependent fashion. The movement of FDP through Egg PC membrane bilayers was confirmed by measuring the conversion of FDP to dihydroxyacetone-phosphate and the formation of hydrozone. FDP (0, 0.5, 5 or 50 mM) was encapsulated in Egg PC multilamellar vesicles and placed in a solution containing aldolase. In the 5 and 50 mM FDP groups there was a significant increase in dihydroxyacetone/hydrazone indicating that FDP crossed the membrane bilayer intact. We theorized that the passive diffusion of FDP might be due to disruption of the membrane bilayer. To examine this hypothesis, small unilamellar vesicles composed of Egg PC were created in the presence of 60 mM carboxyfluorescein, and the leakage of the sequestered dye was followed upon addition of various concentrations of FDP, fructose, fructose-6-phosphate, or fructose-1-phosphate (0, 5 or 50 mM). These results indicate that increasing concentrations of FDP increase the leakage rate of carboxyfluorescein. In contrast, no concentration of fructose, fructose-6-phosphate, or fructose-1-phosphate resulted in any significant increase in membrane permeability to carboxyfluorescein. To examine whether FDP could pass through cellular membranes, we examined the uptake of 14C-FDP by endothelial cells cultured under hypoxia or normoxia for 4 or 16 h. The uptake of FDP was dose-dependent in both the normoxia and hypoxia treated cells, and was accompanied by no significant loss in endothelial cell viability. Our results demonstrate that FDP can diffuse through membrane bilayers in a dose-dependent manner. 相似文献
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207.
森林流域土壤饱和渗透系数与有效孔隙度模型的研究 总被引:10,自引:7,他引:3
以长白山自然保护区内未受扰动的两种典型原始森林土壤为对象,通过野外选取原状土样,在实验室内测定其饱和渗透系数和有效孔隙度,利用回归分析法分别建立了森林土壤内饱和渗透系数和有效孔隙度随深度变化的数学模型,即Ks(z)=K0-f1lna1z和ω(z)=ω0-f2lna2z.将所得出的对数模型与Beven所提出的指数模型进行比较表明,在森林流域内对数模型不仅适用范围比指数模型广,而且精度明显高于指数模型,它更接近于野外实际情况. 相似文献
208.
Nguyen Quynh Thi Kozai Toyoki Niu Genhua Nguyen Uyen Van 《Plant Cell, Tissue and Organ Culture》1998,55(2):133-139
The photosynthetic characteristics of coffee ( Coffea arabusta) plantlets cultured in vitro in response to different CO2 concentrations inside the culture vessel and photosynthetic photon flux (PPF) were investigated preliminarily. The estimation
of net photosynthetic rate (Pn) of coffee plantlets involved three methods: (1) estimating time courses of actual Pn in situ based on measuring CO2 concentrations inside and outside the vessel during a 45-day period, (2) estimating Pn in situ at different CO2 concentrations and PPFs using the above measuring approach for 10-day and 30-day old in vitro plantlets, and (3) estimating
Pn of a single leaf at different CO2 concentrations and PPFs by using a portable photosynthesis measurement system for 45-day old in vitro coffee plantlets. The
results showed that coffee plantlets in vitro had relatively high photosynthetic ability and that the Pn increased with the increase in CO2 concentration inside the vessel. The CO2 saturation point of in vitro coffee plantlets was high (4500–5000 μmol mol-1); on the other hand, the PPF saturation point was not so high as compared to some other species, though it increased with
increasing CO2 concentration inside the vessel.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
209.
210.
Yong-Chun Li ;Jiang-Ping Ren ;Myeong-Je Cho ;Su-Mei Zhou ;Yong-Bum Kim ;Hong-Xiang Guo ;Joshua H. Wong ;Hong-Bin Niu ;Hyun-Kyung Kim ;Susumu Morigasaki ;Peggy G. Lemaux ;Oscar L. Frick ;Jun Yin ;Bob B. Buchanan 《植物生理学报》2009,(3):430-441
Work with cereals (barley and wheat) and a legume (Medicago truncatula) has established thioredoxin h (Trx h) as a central regulatory protein of seeds. Trx h acts by reducing disulfide (S-S) groups of diverse seed proteins (storage proteins, enzymes, and enzyme inhibitors), thereby facilitating germination. Early in vitro protein studies were complemented with experiments in which barley seeds with Trx h overexpressed in the endosperm showed accelerated germination and early or enhanced expression of associated enzymes (α-amylase and pullulanase). The current study extends the transgenic work to wheat. Two approaches were followed to alter the expression of Trx h genes in the endosperm: (1) a hordein promoter and its protein body targeting sequence led to overexpression of Trx hS, and (2) an antisense construct of Trx h9 resulted in cytosolic underexpression of that gene (Arabidopsis designation). Underexpression of Trx h9 led to effects opposite to those observed for overexpression Trx h5 in barley--retardation of germination and delayed or reduced expression of associated enzymes. Similar enzyme changes were observed in developing seeds. The wheat lines with underexpressed Trx showed delayed preharvest sprouting when grown in the greenhouse or field without a decrease in final yield. Wheat with overexpressed Trx h5 showed changes commensurate with earlier in vitro work: increased solubility of disulfide proteins and lower aUergenicity of the gliadin fraction. The results are further evidence that the level of Trx h in cereal endosperm determines fundamental properties as well as potential applications of the seed. 相似文献