BKCa-Cav channel complexes mediate rapid and localized Ca2+-activated K+ signaling

Large-conductance calcium- and voltage-activated potassium channels (BKCa) are dually activated by membrane depolarization and elevation of cytosolic calcium ions (Ca2+). Under normal cellular conditions, BKCa channel activation requires Ca2+ concentrations that typically occur in close proximity to Ca2+ sources. We show that BKCa channels affinity-purified from rat brain are assembled into macromolecular complexes with the voltage-gated calcium channels Cav1.2 (L-type), Cav2.1 (P/Q-type), and Cav2.2 (N-type). Heterologously expressed BKCa-Cav complexes reconstitute a functional "Ca2+ nanodomain" where Ca2+ influx through the Cav channel activates BKCa in the physiological voltage range with submillisecond kinetics. Complex formation with distinct Cav channels enables BKCa-mediated membrane hyperpolarization that controls neuronal firing pattern and release of hormones and transmitters in the central nervous system.     

Respiration and parturition affected by conditional overexpression of the Ca2+-activated K+ channel subunit, SK3

In excitable cells, small-conductance Ca2+-activated potassium channels (SK channels) are responsible for the slow after-hyperpolarization that often follows an action potential. Three SK channel subunits have been molecularly characterized. The SK3 gene was targeted by homologous recombination for the insertion of a gene switch that permitted experimental regulation of SK3 expression while retaining normal SK3 promoter function. An absence of SK3 did not present overt phenotypic consequences. However, SK3 overexpression induced abnormal respiratory responses to hypoxia and compromised parturition. Both conditions were corrected by silencing the gene. The results implicate SK3 channels as potential therapeutic targets for disorders such as sleep apnea or sudden infant death syndrome and for regulating uterine contractions during labor.     

Hyperpolarizing vasodilators activate ATP-sensitive K+ channels in arterial smooth muscle

Vasodilators are used clinically for the treatment of hypertension and heart failure. The effects of some vasodilators seem to be mediated by membrane hyperpolarization. The molecular basis of this hyperpolarization has been investigated by examining the properties of single K+ channels in arterial smooth muscle cells. The presence of adenosine triphosphate (ATP)-sensitive K+ channels in these cells was demonstrated at the single channel level. These channels were opened by the hyperpolarizing vasodilator cromakalim and inhibited by the ATP-sensitive K+ channel blocker glibenclamide. Furthermore, in arterial rings the vasorelaxing actions of the drugs diazoxide, cromakalim, and pinacidil and the hyperpolarizing actions of vasoactive intestinal polypeptide and acetylcholine were blocked by inhibitors of the ATP-sensitive K+ channels, suggesting that all these agents may act through a common pathway in smooth muscle by opening ATP-sensitive K+ channels.     

Cytosolic calcium during contraction of isolated mammalian gastric muscle cells

During activation of visceral smooth muscle there is an increase in cytosolic-free calcium, but the source (intracellular calcium release or calcium influx), kinetics, and stoichiometry of this increase have not been determined. Here, the fluorescent indicator, quin2-acetoxymethyl ester, was used to measure directly cytosolic-free calcium during contraction of isolated stomach muscle cells induced by the two neuropeptides cholecystokinin-octapeptide and Met-enkephalin as well as acetylcholine. An increase in cytosolic-free calcium was seen that was (i) dependent on the concentration of contractile agonist, (ii) derived from intracellular sources (that is, not significantly affected by removal of ambient calcium or addition of a calcium channel blocker), and (iii) kinetically and stoichiometrically related to net calcium efflux and contraction. In contrast, the increase in cytosolic-free calcium induced by depolarizing concentrations of potassium was caused by influx of calcium through voltage-dependent calcium channels.     

Alteration of four identified K+ currents in Drosophila muscle by mutations in eag

Voltage-clamp analysis of Drosophila larval muscle revealed that ether à go-go (eag) mutations affected all identified potassium currents, including those specifically eliminated by mutations in the Shaker or slowpoke gene. Together with DNA sequence analysis, the results suggest that the eag locus encodes a subunit common to different potassium channels. Thus, combinatorial assembly of polypeptides from different genes may contribute to potassium channel diversity.     

Relaxation of isolated gastric smooth muscle cells by vasoactive intestinal peptide

Vasoactive intestinal peptide caused a prompt, dose-dependent relaxation of isolated gastric smooth muscle cells of the guinea pig and a significant increase in intracellular adenosine 3',5'-monophosphate coincidentally with optimum relaxation. Relaxation was augmented by a threshold concentration of isobutyl methylxanthine. The direct relaxant effect of vasoactive intestinal peptide and the distribution of nerves containing this peptide to circular smooth muscle support the view that vasoactive intestinal peptide is the neuromuscular transmitter of enteric inhibitory nerves.     

Ca2+ Fe2+ Mg2+ Zn2+ Na+ K+对哺乳仔猪乳糖酶活力的影响

乳糖和矿质元素是哺乳仔猪十分重要的营养素,对7日龄的哺乳仔猪乳糖酶研究结果表明:哺乳仔猪乳糖酶的酶活力常数是65.825μg(ONP)/mL.m in,矿质元素钠、铁和钙能促进乳糖酶的酶活力。     

Ca2+ Fe2+ Mg2+ Zn2+ Na+ K+对哺乳仔猪乳糖酶活力的影响

乳糖和矿质元素是哺乳仔猪十分重要的营养素,对7日龄的哺乳仔猪乳糖酶研究结果表明:哺乳仔猪乳糖酶的酶活力常数是65.825μg(ONP)/mL·min,矿质元素钠、铁和钙能促进乳糖酶的酶活力.     

盐胁迫条件下杨树组织及细胞中钾、钙、镁的变化

该文研究了NaCl对群众杨(Populus‘popularis 35-44', P. popularis)、I-214杨(P. ×euramericana cv. I-214, P. cv. I-214)和胡杨(P. euphratica)3种杨树组织和细胞营养状况的影响. 为期30 d的盐处理降低了群众杨和I-214杨根和茎组织中K+、Ca2+、Mg2+的水平,而对于叶片营养元素的水平基本没有影响. 与群众杨和I-214杨比较,胡杨组织的营养水平受NaCl的影响最小. 根皮层细胞X-射线微区分析的结果显示,NaCl降低了群众杨和I-214杨细胞壁和液泡中K+、Mg2+的水平,但对于胡杨细胞营养元素水平的影响较小. 实验结果表明,胡杨在盐胁迫条件下能保持对营养元素的吸收,维持较好的营养状况,这是其抗盐性强的原因之一;I-214杨和群众杨的抗盐性弱,与其胁迫条件下营养状况的恶化有关.     

Na+,K+,Mg2+,Ca2+浓度对嗜盐放线菌生长的影响

通过研究近30株嗜盐或耐盐放线菌在不同浓度的Na^＋，K^＋，M^2＋，Ca^2＋条件下的生长范围，发现耐盐放线菌对Na^＋，K^＋，M^2＋有广泛的适应性。只有少数耐盐放线菌能在较底浓度的CaCl2条件下生长；多数嗜盐放线菌生长所需的Na^＋可被K^＋，Mg^2＋所替代，而不能被Ca^2＋所替代，少数嗜盐放线菌的生长离不开Na^＋，对Na^＋有高度的专一性。因此，提出自然环境中是否也存在类似的专嗜K^＋或专嗜Mg^2＋的嗜盐放线菌的推测。     

Cytoprotective role of Ca2+- activated K+ channels in the cardiac inner mitochondrial membrane

Ion channels on the mitochondrial inner membrane influence cell function in specific ways that can be detrimental or beneficial to cell survival. At least one type of potassium (K+) channel, the mitochondrial adenosine triphosphate-sensitive K+ channel (mitoKATP), is an important effector of protection against necrotic and apoptotic cell injury after ischemia. Here another channel with properties similar to the surface membrane calcium-activated K+ channel was found on the mitochondrial inner membrane (mitoKCa) of guinea pig ventricular cells. MitoKCa significantly contributed to mitochondrial K+ uptake of the myocyte, and an opener of mitoKCa protected hearts against infarction.     

Tension transients in single isolated smooth muscle cells

Tension transients were recorded in a single smooth muscle cell. The transient contains a linear elastic response and a biphasic recovery that appear to originate from the cross-bridges. A comparison of transients in smooth and fast skeletal muscle fibers suggests that the cross-bridge in smooth muscle is more compliant than the cross-bridge in striated muscle and that transitions between several cross-bridge states occur more slowly in smooth muscle than in striated muscle.     

Inositol 1,3,4,5-tetrakisphosphate induces Ca2+ sequestration in rat liver cells

Inositol 1,4,5-trisphosphate [I(1,4,5)P3] is a second messenger generated along with diacylglycerol upon the binding of various physiological agents with their cell surface receptors. I(1,4,5)P3 mobilizes Ca2+ from intracellular storage sites through a receptor-coupled mechanism, and the subsequent increased intracellular free calcium ion concentration [( Ca2+]i) activates a multitude of cellular responses. Electropermeabilized neoplastic rat liver epithelial (261B) cells were used to study Ca2+ sequestration, a process that reverses the elevated [Ca2+]i to resting levels and replenishes intracellular Ca2+ pools. Although I (1,4,5)P3-mobilized Ca2+ is readily sequestered into storage pools by the action of Ca2+-adenosine triphosphatases, Ca2+ mobilized by addition of the nonmetabolized inositol trisphosphate isomer I(2,4,5)P3 is not sequestered, suggesting that metabolism is necessary to eliminate the stimulus for Ca2+ release. Several inositol phosphate compounds were examined for their ability to lower the buffer [Ca2+] to determine if a specific I(1,4,5)P3 metabolite might be involved in stimulating Ca2+ sequestration; of these, I(1,3,4,5)P4 alone was found to induce Ca2+ sequestration, demonstrating a physiological role for this inositol trisphosphate metabolite.     

黑土中NH_4~+、K~+、Ca~(2+)、Mg~(2+)对烤烟中钾含量的影响

采用框栽田间模拟试验方法对哈尔滨黑土中不同浓度养分离子对烟叶中钾含量的影响进行了研究。结果表明 ,烟叶中钾含量与土壤中 NH4 +浓度无明显相关性 ,而与土壤中 K+浓度呈显著正相关 γ=0 .935* ,土壤中 Ca2 +、Mg2 +浓度增加使烟叶中钾含量有所降低     

Restoration by calmodulin of a Ca2+-dependent K+ current missing in a mutant of Paramecium

A combination of genetics, biochemistry, and biophysics was used to show that calmodulin is involved in the regulation of an ion channel. Calmodulin restored the Ca2+-dependent K+ current in pantophobiac, a mutant in Paramecium that lacks this current. The restoration of the current occurred within 2 hours after the injection of 1 picogram of wild-type calmodulin into the mutant. The current remained for approximately 30 hours before the mutant phenotype returned. The injection of calmodulin isolated from pantophobiac had no effect. These results imply that calmodulin is required for the function or regulation of the Ca2+-dependent K+ current in Paramecium.     

Platelet-mediated cholesterol accumulation in cultured aortic smooth muscle cells

Cholesterol accumulates within smooth muscle cells and macrophages in atherosclerotic lesions, thereby contributing to the progressive enlargement of these lesions. The mechanism of this cellular accumulation of cholesterol is not known. The possibility that platelets may have a role in the cellular cholesterol accumulation that occurs during atherogenesis was investigated. Incubation of thrombin-activated washed rat platelets (or platelet-free supernatants prepared from thrombin-activated platelets) with cultured rat aortic smooth muscle cells induced cholesteryl ester lipid droplet accumulation within the smooth muscle cells. No cholesteryl ester lipid droplets accumulated when smooth muscle cells were incubated with unactivated platelets. Smooth muscle cell lipid droplet accumulation occurred in the absence of serum lipoproteins and was not inhibited by mevinolin, a drug that blocks cholesterol synthesis. These findings suggest that activated platelets may release cholesterol, which can be accumulated by cells and stored as lipid droplets.     

Regulation of calcium concentration in voltage-clamped smooth muscle cells

The regulation of intracellular calcium concentration in single smooth muscle cells was investigated by simultaneously monitoring electrical events at the surface membrane and calcium concentration in the cytosol. Cytosolic calcium concentration rose rapidly during an action potential or during a voltage-clamp pulse that elicited calcium current; a train of voltage-clamp pulses caused further increases in the calcium concentration up to a limit of approximately 1 microM. The decline of the calcium concentration back to resting levels occurred at rates that varied with the calcium concentration in an apparently saturable manner. Moreover, the rate of decline at any given calcium concentration was enhanced after a higher, more prolonged increase of calcium. The process responsible for this enhancement persisted for many seconds after the calcium concentration returned to resting levels. Thus, the magnitude and duration of a calcium transient appear to regulate the subsequent calcium removal.     

Ca2+和K+对紫椴叶片渗透调节指标的影响

采用不同质量分数的Ca2 和K 溶液处理2年生紫椴幼苗,在-10℃低温条件下分别处理4、6、8、10h,测定和分析其质膜相对透性、丙二醛(MDA)浓度、可溶性糖质量分数及游离脯氨酸质量分数的变化。结果表明:不同质量分数Ca2 和K 的处理能够降低质膜透性、减轻低温胁迫对细胞膜的破坏、减少丙二醛的浓度、增加可溶性糖和游离脯氨酸质量分数。其中,经过K 处理的植株抵御低温效果显著,并以质量分数为250%的处理效果最佳。     

Cu、Zn对烤烟幼苗吸收K、Ca、Fe的影响

研究了溶液培养条件下Cu．Zn对烟草幼苗吸收K．Ca、Fe的影响。结果表明，烟草幼苗对Cu．Zn的吸收随溶液中Cu^2 、Zn^2 浓度的升高而增加，Zn对Cu的吸收起抑制作用。Ca吸收随溶液Cu^2 浓度的升高而呈下降趋势，而随溶液Zn^2 浓度的升高而有增加趋势；K吸收随溶液中Cu^2 、Zn^2 浓度的增大在茎叶中有升高趋势，而在根部有下降趋势；幼苗对于Fe吸收随溶液中Cu^2 、Zn^2 浓度的增大而略有增加，但差异不显著。     

菲和芘胁迫对平邑甜茶根毛细胞钙、钾和氢离子流动性的影响

【目的】探明多环芳烃(polycyclic aromatic hydrocarbons,PAHs)对植物生长毒害的机理,以及PAHs中不同物质毒害效应的差异。【方法】采用非损伤微测技术测定平邑甜茶幼苗根毛经多环芳烃菲和芘处理后Ca2+、 K+、H+流速变化。【结果】经过菲处理后,Ca2+、K+、H+流动性发生明显逆转,平均流速由对照的（-63.53±9.30） pmol•cm-2•s-1、（-60.56±14.56） pmol•cm-2•s-1、（+44.38±5.19 ） pmol•cm-2•s-1分别改变为(+127.18±39.95) pmol•cm-2•s-1 、(+109.97±25.68) pmol•cm-2•s-1、(-10.35±1.57 ) pmol•cm-2•s-1。经芘处理后,Ca2+、K+、H+流动性同样发生逆转,平均流速分别改变为（+220.29±60.42） pmol•cm-2•s-1、（+140.21±27.87） pmol•cm-2•s-1)、（-14.42±3.16） pmol•cm-2•s-1。【结论】PAHs破坏了细胞膜通透性,降低了离子通道活性,活化了根细胞质膜 Ca2+-ATPase 及 Ca2+/ H+反向转运体,扰乱了平邑甜茶根系对Ca2+、K+离子的吸收过程,造成相应元素缺失,并且芘造成的毒害效应显著高于菲,成为PAHs毒害植物体的机理之一。说明多环芳烃对植物生长的影响可以追溯到离子吸收及离子通道的改变上,为进一步研究PAHs对植物影响提供了理论依据。