Mechanism of inhibition of net ion transport across frog corneal epithelium by calcium channel antagonists

Summary In the isolated bullfrog cornea, three calcium channel antagonists had dose-dependent inhibitory effects on the Cl-originated short-circuit current (SCC). Their order of decreasing potency was bepridil, verapamil and diltiazem. One millimolar diltiazem inhibited the SCC by 98% and subsequent incubation with the calcium ionophore A23187 had no restorative effect. Increasing the bathing solution Ca concentration from 0.05 to 15mm, however, decreased diltiazem's inhibitory efficacy. This antagonist depolarized the intracellular potential differenceV _m from –54 to –18 mV (tear: reference) and the voltage divider ratioFR ₀ decreased from 0.58 to 0.30, suggesting an increase in basolateral membrane electrical resistance. Additional indication of a basolateral membrane effect by the drug was that preincubation with 10⁵ m amphotericin B in Cl-free Ringer's did not eliminate the inhibitory effect of the drug on the Na- and K-elicited SCC. In the absence of amphotericin B in Cl-free Ringer's (SCC=0), 1 ×10³ m diltiazem depolarized theV _m from –78 to –9 mV suggesting that the increase in basolateral membrane resistance was due to K channel blockade. Diltiazem (1×10³ m) significantly decreased cyclic AMP content; however, isoproterenol in the presence of the drug increased cyclic AMP fourfold without having any restorative effect on the inhibited SCC. Therefore, the inhibition of the Cl-originated SCC resulting from an increase in basolateral membrane K resistance is not caused by a decline in cyclic AMP content. In plasma membrane-enriched fractions prepared from broken cell preparations of bovine corneal epithelium, 1×10³ m diltiazem had no inhibitory effects on either Na,K-ATPase or Ca,Mg-ATPase activities. These latter effects further point to the selectivity of diltiazem as an inhibitor of K-channel activity, but do not preclude a Ca-channel blocker effect by the drug in the micromolar range.     

Single-channel analysis of a potassium inward rectifier in myocytes of newborn rat heart

Summary Unitary K⁺ currents in single cells isolated from ventricular muscle of newborn rat hearts were measured in response to different potentials and [K] _o . TheI/V curves were linear for potentials more negative than the zero-current voltage: especially in high [K] _o (150nm KCl), no clear outward currents could be detected indicating a drastic rectification in the inward direction. The channel is mainly selective to K⁺ but Na⁺ ions are also carried (P _Na/P _K=0.056). The channel conductance is proportional to the square root of [K] _o but Na⁺ ions seem to have a facilitatory effect on _K, the single-channel conductance. The channel activity, measured asP _o, i.e. the probability to find the channel in open state, decreased as the membrane was hyperpolarized. This behavior was tentatively explained by an inactivation process as the membrane becomes more negative. The rate constants of the transitions between the different states were calculated according to a C–O–C model. A control of the gating process by permeant ion K⁺ was postulated, based on the increase of one of the rate constants from the closed to the open state with [K] _o . Finally, the macroscopicI/V curves calculated fromP _o and i, the unit current, were found to be characteristic of a ion-blocked inward rectifier.     

Effects of saponin extracts from Solanum elaeagnifotium fruits on ion uptake by clover seedlings

Solanum elaeagnifolium Cav. fruits contain high concentrations of steroidal saponins. Treatment of 3-day-old clover seedlings with aqueous fruit extracts modified Ca²⁺ uptake without significantly altering K⁺ and H₂PO₄⁻ uptake. The extracts increased Ca²⁺ uptake in the concentration range of 0.2 to 20 m M Ca²⁺. Uptake curves could be represented by two phases. In the lower phase (0.2-1.0 m M Ca²⁺), this change could be related to an increase in V_max. Pretreatment of seedlings with saponin extracts significantly reduced ATP-dependent Ca²⁺ uptake and Ca²⁺-dependent ATPase activity in a fraction isolated from root homogenates by centrifugation at 1500 g for 15 min. Saponins purified from S. eleagnifolium extracts by thin-layer chromatography modified in vitro the Ca²⁺-ATPase activity of this fraction, indicating that the steroid may act directly on Ca²⁺ transport across membranes.     

Effects of monovalent cations on red cell shape and size

Human erythrocytes were incubated in isotonic solutions of different monovalent cations. The apparent size of the red cells measured on scanning electron microscopic pictures decreases in the order Li⁺>Na⁺=K⁺>Rb⁺. These differences in size are abolished after pretreatment with trypsin, which removes a large part of the charges associated with membrane glycoproteins. Shape alterations are also observed. Normal biconcave shapes are visible after Na⁺ or K⁺ incubation, whereas Li⁺ leads to flabby, flattened cells with a certain tendency to crenation, and Rb⁺ causes more pronounced biconcavity with a certain tendency to cupping. The overall effects of pretreatment with trypsin are similar to those of Li⁺. Our results provide evidence that the electrostatic repulsion of glycoproteins and other charged membrane components may play an essential role in maintaining red cell shape.     

脯氨酸对缺钾植株的保护作用

在缺钾培养条件下,高粱苗地上部单位鲜重含钾量由于地上部鲜重的增加而下降。从高梁苗钾缺乏症出现的时间、频率和程度来看,Pro培养似乎有利于缓和或减轻缺钾对植物造成的伤害。在同样含钾量情况下,以Ca(NO_3)_2为氮源的苗受到缺钾伤害要重于以Pro为氮源的苗受到的伤害,其钾缺乏症植株的百分数增加值与含钾量降低值之比为0.26,以Pro为氮源的缺钾营养液培养的苗为0.09,即下降同样量的钾前者苗的钾缺乏症的增加要大大地超过后者。Pro并不减轻在缺钾情况下叶绿素含量的下降。     

Regulation of the basolateral potassium conductance of theNecturus proximal tubule

Summary Two methods, the measurement of the response of the basolateral membrane potential (V _bl) of proximal tubule cells ofNecturus to step changes in basolateral K⁺ concentration, and cellular cable analysis, were used to assess the changes in basolateral potassium conductance (G _K) caused by a variety of maneuvers. The effects of some of these maneuvers on intracellular K⁺ activity (a _K ⁱ ) were also evaluated using double-barreled ion-selective electrodes. Perfusion with 0mm K⁺ basolateral solution for 15 min followed by 45 min of 1mm K⁺ solution resulted in a fall in basolateral potassium (apparent) transference number (t _K),V _bl anda _K ⁱ . Results of cable analysis showed that total basolateral resistance,R _b , rose. The electrophysiological effects of additional manipulations, known to inhibit net sodium reabsorption across the proximal tubular epithelium ofNecturus, were also investigated. Ouabain caused a fall int _K accompanied by large decreases ina _K ⁱ andV _bl. Lowering luminal sodium caused a fall int _K and a small reduction inV _bl. Selective reduction of peritubular sodium, a maneuver that has been shown to block sodium transport from lumen to peritubular fluid, also resulted in a significant decrease int _K. These results suggest thatG _K varies directly with rate of transport of the sodium pump, irrespective of the mechanism of change in pump turnover.Part of this material has been presented at the 10th International Conference on Biological Membranes (Cohen & Giebisch, 1984).     

Current-voltage relations of the basolateral membrane in tight amphibian epithelia: Use of nystatin to depolarize the apical membrane

Summary Exposure of the mucosal side of toad(Bufo bufo) urinary bladder and frog(Rana ridibunda) skin to the polyene ionophore nystatin, resulted in stable preparations in which the apical resistance was negligible compared to the basolateral resistance. The preparations support passive K currents in both directions and an amiloride-insensitive Na current in the apicalserosal direction which is blocked by ouabain. The nystatintreated toad bladder was used to study the electrical properties of the basolateral membrane by means of current-voltage curves recorded transepithelially. The K current showed strong rectification at cellular potentials negative with respect to the interstitial space. The ouabain-sensitive current increased with membrane voltage at negative voltages but saturated above+20 mV.     

Effects of external NaCl on the growth of Atriplex amnicola and the ion relations and carbohydrate status of the leaves

Abstract Atriplex amnicola, was grown in nutrient solution cultures with concentrations of NaCl up to 750 mol m^?3. The growth optimum was at 25–50 mol m^?3 NaCl and growth was 10–15% of that value at 750 mol m^?3 NaCl. Sodium chloride at 200 mol m^?3 and higher reduced the rate of leaf extension and increased the time taken for a leaf to reach its maximal length. Concentrations of Na⁺, K⁺ and Mg²⁺ in leaves of different ages were investigated for plants grown at 25, 200 and 400 mol m^?3 NaCl. Although leaves of plants grown at 200 and 400 mol m^?3 NaCl had high Na⁺ concentrations at young developmental stages, much of this Na⁺ was located in the salt bladders. Leaves excluding bladders had low Na⁺ concentrations when young, but very high in Na⁺ when old. In contrast to Na⁺, K⁺ concentrations were similar in bladders and leaves excluding bladders. Concentrations of K⁺ were higher in the rapidly expanding than in the old leaves. At 400 mol m^?3 NaCl, the K⁺:Na⁺ ratios of the leaves excluding bladders were 0.4–0.6 and 0.1 for rapidly expanding and oldest leaves, respectively. The Na⁺ content in moles per leaf, excluding bladders, increased linearly with the age of the leaves; concurrent increases in succulence were closely correlated with the Na ⁺ concentration in the leaves excluding the bladders. Soluble sugars and starch in leaves, stems and buds were determined at dusk and dawn. There was a pronounced diurnal fluctation in concentrations of carbohydrates. During the night, most plant parts showed large decreases in starch and sugar. Concentrations of carbohydrates in most plant organs were similar for plants grown at 25 and 400 mol m^?3 NaCl. One notable exception was buds at dusk, where sugar and starch concentrations were 30–35% less in plants grown at 400 mol m^?3 NaCl than in plants grown at 25 mol m^?3 NaCl. The data indicate that the growth of A. amnicola at 400 mol m^?3 NaCl is not limited by the availability of photosynthate in the plant as a whole. However, there could have been a growth limitation due to inadequate organic solutes for osmotic regulation.     

Accumulation and subcellular distribution of cations in relation to the growth of potassium-deficient barley

Abstract Growth of barley (Hordeum vulgare L., cv. Georgie) was insensitive to soil K content above about 150 mg kg^?1, but at lower levels it declined. The reduction in yield was greater in soils containing approximately 10 mg Na kg^?1 than in soils with about 90 mg kg^?1 of Na. Growth was unaffected by changes in shoot K concentration above 75 mol m^?3, but declined at lower concentrations, and the decrease was less in plants grown in soils with high Na. Growth responses were not simply related to tissue K concentrations because plants grown in soils with extra Na had higher yields but lower K concentrations. When soil Na was low, plants accumulated Ca as tissue K declined, but when Na was provided this ion was accumulated. Plant Mg concentrations were generally low but increased as K decreased. The Ca and Mg were osmotically active. There were highly significant inverse linear relationships between yield and either the Ca or Mg concentrations in the shoots. X-ray microanalysis was used to examine the compartmentation of cations in leaves from barley plants (cv. Clipper) grown in nutrient solutions with high and low K concentrations. In plants grown with 2.5 mol m^?3 K, this was the major cation in both the cytoplasm and vacuole of mesophyll cells. However, in plants grown with 0.02 mol m^?3 K it declined to undetectable levels in the vacuole, although it was still detectable in the cytoplasm. In all plants, Ca was mainly located in epidermal cells. The implication of the results for explaining responses to K. in terms of compartmentation of solutes is discussed.     

窦性节律下家兔心率与在位心室肌纤维有效不应期的关系

在自然呼吸和窦性节律下,用浮置式玻璃微电极引导在体单个左心室肌纤维动作电位,作为兴奋的指标,以其 0相触发产生期前的试验刺激,测定有效不应期(ERP)。38只家兔的测定结果表明,在R-R间期为205—330ms的范围内,随着心率加快(R-R间期缩短),ERP减小,而 ERP/RR 间期增大,说明 ERP 与心率直接有关。并且,在较快的心率时,ERP 相对延长。通过相关与回归分析,制出了能够删除心率影响的校正公式。静脉注射酒石酸锑钾(50mg/kg)发现,在窦性和起搏节律下,酒石酸锑钾均能轻度延长 ERP(P<0.001)。窦性节律下的校正后值与起搏节律的测定结果完全一致。证明校正后值能够用来比较处理前后不同心率条件下,各种药物、离子及其它因素对ERP 的影响。 本文的校正公式虽然只适用于同种动物和方法,但此校正公式的制作原理也可以广泛应用到其它多种动物。