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1.
Single calcium-dependent potassium channels in clonal anterior pituitary cells. 总被引:9,自引:1,他引:9 下载免费PDF全文
Single Ca2+-dependent K+-channel currents were recorded in intact and excised inside-out membrane patches of the anterior pituitary clone AtT-20/D16-16. The frequency of channel openings and lifetimes depends both on membrane potential and on the Ca2+ concentrations at the inner membrane surface. The curve of the open-state probability of the channel as a function of membrane potential appears to translate along the voltage axis with changes in internal Ca2+ concentration. For Ca2+ concentrations between 10(-7) and 10(-6) M, the shift is consistent with the hypothesis that three Ca2+ ions are required to open a Ca2+-dependent K+ channel. Single channel conductances are estimated to be 124 pS in patches with normal external K+ (5.4 mM) and 208 pS in excised patches with symmetrical K+ (145 mM) across the membrane. Tetraethylammonium (20 mM) added to the cytoplasmic surface reversibly blocks the Ca2+-dependent K+ channel. 相似文献
2.
Calcium-activated potassium channels in liver cells 总被引:1,自引:0,他引:1
Activation of certain membrane receptors increases the concentration of Ca2+ in the cytosol of hepatocytes. Since in most species these cells possess a PK(Ca) mechanism, the outcome is a rise in PK. This can be blocked by quinine, apamin and certain neuromuscular blocking agents. The binding of labelled apamin to hepatocytes has been studied under physiological conditions, and the relationship between the binding sites and K+ channels is discussed. The physiological role of the PK(Ca) mechanism in hepatocytes is unclear, though it is largely responsible for ‘adrenaline hyperkalaemia’. 相似文献
3.
Dexamethasone increases potassium channel messenger RNA and activity in clonal pituitary cells. 总被引:5,自引:0,他引:5
E S Levitan L M Hemmick N C Birnberg L K Kaczmarek 《Molecular endocrinology (Baltimore, Md.)》1991,5(12):1903-1908
Glucocorticoid hormones are released as part of the stress response and regulate secretion by the pituitary. Since the activity of ion channels also influences secretion, we examined the effect of the glucocorticoid agonist dexamethasone on ion channel expression. K+ channel mRNA was detected in rat hypothalamus and anterior pituitary, with probes derived from the rat Kv1 gene, a member of the mammalian voltage-gated K+ channel superfamily. High levels were also detected in PRL-secreting clonal (GH3 and GH4C1) rat pituitary cells. Dexamethasone rapidly increased the steady state concentration of Kv1 mRNA in GH3 cells in a dose-dependent manner. This change in gene expression was accompanied by an increase in whole cell voltage-gated K+ current [lk(i)] with similar pharmacology to the Kv1 gene product. Our findings indicate that hormones may act directly on excitable cells to produce long term effects on electrical activity and secretion by regulating K+ channel expression. 相似文献
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6.
Calcium-activated potassium channels in chondrocytes. 总被引:2,自引:0,他引:2
M Grandolfo P D'Andrea M Martina F Ruzzier F Vittur 《Biochemical and biophysical research communications》1992,182(3):1429-1434
The presence of calcium-activated potassium channels in chondrocytes of growing cartilage was tested. Results obtained with fura-2 on cultured resting chondrocytes indicate that the cells respond to an elevation of extracellular calcium concentration ([Ca2+]o) from 0.1 to 2 mM increasing the intracellular concentration of the ion ([Ca2+]i) from 117 to 187 nM. This increment may be blocked by 3 microM La3+. Patch clamp experiments in cell-attached configuration showed that, when [Ca2+]i rises, the open probability (Po) of the K+ channels increases. Increments in both Po and unitary currents of the K+ channels can be obtained after applying 2.5 microM A23187 with 2 mM [Ca2+]o. Hence, the results demonstrate that, in chondrocytes, a class of Ca(2+)-activated K+ channels is present and their activity is related to an increase of [Ca2+]i. 相似文献
7.
Linear impedance studies of voltage-dependent conductances in tissue cultured chick heart cells. 下载免费PDF全文
Plateau and pacemaker currents from tissue cultured clusters of embryonic chick heart cells were studied in the time domain, using voltage-clamp steps, and in the frequency domain, using a wide-band noise input superimposed on a steady holding voltage. In the presence of tetrodotoxin to block the sodium channel, a depolarizing voltage step into the plateau range elicited: (a) a rapid (approximately equal to 2 ms) activation of the slow inward current; (b) a subsequent slower (approximately equal to 25 ms) decline in the slow inward current; and (c) activation of a very slow (5 to 10 s) outward current. Impedance studies in this voltage range could clearly resolve two voltage-dependent processes, which appeared to correspond to points b and c above because of their voltage dependence, pharmacology, and time constants. A correlate of point a was also probably present but difficult to resolve owing to the fast time constant of activation for the slow inward channel. At voltages negative to -50 mV a new voltage-dependent process could be resolved, which, because of its voltage dependence and time constant, appeared to represent the pacemaker channel (also termed If or IK2). In the Appendix, linear models of voltage-dependent channels and ion accumulation/depletion are derived and these are compared with our data. Most of the above-mentioned processes could be attributed to voltage-dependent channels with kinetics similar to those observed in time domain, voltage-clamp studies. However, the frequency domain correlate of the decline of the slow inward current was incompatible with channel gating, rather, it appears accumulation/depletion of calcium may dominate the decline in this preparation. 相似文献
8.
Calcium-activated potassium channels in human platelets 总被引:1,自引:0,他引:1
B P Fine K A Hansen J R Salcedo A Aviv 《Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.)》1989,192(2):109-113
The cationic fluorescent probe, DiSC3(5) was used to measure the membrane potential in human platelets. Hyperpolarization was induced by the addition of Ca2+ to the medium and also by the addition of the Ca2+ ionophore, A23187. In the absence of extracellular Ca2+ ([Ca2+]o) there was no response to A23187. The threshold concentration for [Ca2+]o was 20 microM and for A23187 was 12 nM. The increase polarity induced by [Ca2+]o was not affected by various K+ channel blockers. However, the effect of A23187 was inhibited by quinine and charybdotoxin, while apamin, tetraethylammonium, and the calmodulin inhibitors trifluoperazine and compound R24571 were ineffective. The resting membrane potential was -66 +/- 0.9 mV and was decreased by quinine. There are three conclusions from this study: (i) Ca2+-activated K+ channels exist in human platelets; (ii) they are the type that are apamin insensitive, charybdotoxin sensitive; and (iii) they may contribute to the resting membrane potential. 相似文献
9.
W E Crill 《Journal of Physiology》1999,93(1-2):17-21
Layer five pyramidal neurons of rat and cat neocortex have numerous ionic conductance mechanisms. The presence of these voltage-dependent conductances in the dendrites has a significant effect on the transmission of current from synaptic sites to the spike generating region in the proximal axon. Here we show such threshold activation of persistent sodium channels markedly amplifies current flowing through glutamate activated dendritic channels. 相似文献
10.
Functional significance of voltage-dependent conductances in Limulus ventral photoreceptors 下载免费PDF全文
《The Journal of general physiology》1982,79(2):211-232
The influence of voltage-dependent conductances on the receptor potential of Limulus ventral photoreceptors was investigated. During prolonged, bright illumination, the receptor potential consists of an initial transient phase followed by a smaller plateau phase. Generally, a spike appears on the rising edge of the transient phase, and often a dip occurs between the transient and plateau. Block of the rapidly inactivating outward current, iA, by 4-aminopyridine eliminates the dip under some conditions. Block of maintained outward current by internal tetraethylammonium increases the height of the plateau phase, but does not eliminate the dip. Block of the voltage-dependent Na+ and Ca2+ current by external Ni2+ eliminates the spike. The voltage-dependent Ca2+ conductance also influences the sensitivity of the photoreceptor to light as indicated by the following evidence: depolarizing voltage- clamp pulses reduce sensitivity to light. This reduction is blocked by removal of external Ca2+ or by block of inward Ca2+ current with Ni2+. The reduction of sensitivity depends on the amplitude of the pulse, reaching a maximum at or approximately +15 mV. The voltage dependence is consistent with the hypothesis that the desensitization results from passive Ca2+ entry through a voltage-dependent conductance. 相似文献
11.
Calcium-activated potassium conductance noise in snail neurons 总被引:1,自引:0,他引:1
Current fluctuations were measured in small, 3-6 micrometers-diameter patches of soma membrane in bursting neurons of the snail, Helix pomatia. The fluctuations dramatically increased in magnitude with depolarization of the membrane potential under voltage clamp conditions. Two components of conductance noise were identified in the power spectra calculated from the membrane currents. One component had a corner frequency which increased with depolarization. This component was blocked by intracellular injection of TEA and was relatively insensitive to extracellular calcium levels (as long as the total number of effective divalent cations remained constant). It was identified as fluctuations of the voltage-dependent component of delayed outward current. The second component of conductance noise had a corner frequency which decreased with depolarization. It was relatively unaffected by TEA injection and was reversibly blocked by substitution of extracellular calcium with magnesium, cobalt, or nickel. This second component of noise was identified as fluctuations of the calcium-dependent potassium current. The results suggest that the two components of delayed outward current are conducted through physically distinct channels. 相似文献
12.
Yuko Kaneko Takehiko Saito 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1992,170(4):411-425
Summary Electrical membrane properties of solitary spiking cells during newt (Cynops pyrrhogaster) retinal regeneration were studied with whole-cell patch-clamp methods in comparison with those in the normal retina.The membrane currents of normal spiking cells consisted of 5 components: inward Na+ and Ca++ currents and 3 outward K+ currents of tetraethylammonium (TEA)-sensitive, 4-aminopyridine (4-AP)-sensitive, and Ca++-activated varieties. The resting potential was about -40mV. The activation voltage for Na+ and Ca++ currents was about -30 and -17 mV, respectively. The maximum Na+ and Ca++ currents were about 1057 and 179 pA, respectively.In regenerating retinae after 19–20 days of surgery, solitary cells with depigmented cytoplasm showed slowrising action potentials of long duration. The ionic dependence of this activity displayed two voltage-dependent components: slow inward Na+ and TEA-sensitive outward K+ currents. The maximum inward current (about 156 pA) was much smaller than that of the control. There was no indication of an inward Ca++ current.During subsequent regeneration, the inward Ca++ current appeared in most spiking cells, and the magnitude of the inward Na+, Ca++, and outward K+ currents all increased. By 30 days of regeneration, the electrical activities of spiking cells became identical to those in the normal retina. No significant difference in the resting potential and the activation voltage for Na+ and Ca++ currents was found during the regenerating period examined. 相似文献
13.
Calcium-activated potassium channels: Regulation by calcium 总被引:28,自引:0,他引:28
Owen B. McManus 《Journal of bioenergetics and biomembranes》1991,23(4):537-560
14.
Rosenfeld CR White RE Roy T Cox BE 《American journal of physiology. Heart and circulatory physiology》2000,279(1):H319-H328
Nitric oxide synthase (NOS) contributes to estradiol-17beta (E(2)beta)-induced uterine vasodilation, but additional mechanisms are involved, and the cellular pathways remain unclear. We determined if 1) uterine artery myocytes express potassium channels, 2) E(2)beta activates these channels, and 3) channel blockade plus NOS inhibition alters E(2)beta-induced uterine vasodilation. Studies of cell-attached patches identified a 107 +/- 7 pS calcium-dependent potassium channel (BK(Ca)) in uterine artery myocytes that rapidly increased single-channel open probability 70-fold (P < 0.05) after exposure to 100 nM E(2)beta through an apparent cGMP-dependent mechanism. In ovariectomized nonpregnant ewes (n = 11) with uterine artery flow probes and catheters, local BK(Ca) blockade with tetraethylammonium (TEA; 0.05-0.6 mM) dose dependently inhibited E(2)beta-induced uterine vasodilation (n = 37, R = 0.77, P < 0.0001), with maximum inhibition averaging 67 +/- 11%. Mean arterial pressure (MAP) and E(2)beta-induced increases (P = 0.001) in heart rate (13%) and contralateral uterine blood flow (UBF, approximately 5-fold) were unaffected. Local NOS inhibition plus BK(Ca) blockade, using submaximal doses of nitro-L-arginine methyl ester (5 mg/ml) and TEA (0.3 mM), did not alter basal UBF but completely inhibited ipsilateral E(2)beta-induced uterine vasodilation without affecting MAP and E(2)beta-induced increases in contralateral UBF and heart rate. Acute E(2)beta-mediated uterine vasodilation involves rapid activation of uterine artery BK(Ca) and NOS, and the pathway for their interaction appears to include activation of guanylyl cyclase. 相似文献
15.
Properties of calcium and potassium currents of clonal adrenocortical cells 总被引:1,自引:2,他引:1 下载免费PDF全文
The ionic currents of clonal Y-1 adrenocortical cells were studied using the whole-cell variant of the patch-clamp technique. These cells had two major current components: a large outward current carried by K ions, and a small inward Ca current. The Ca current depended on the activity of two populations of Ca channels, slow (SD) and fast (FD) deactivating, that could be separated by their different closing time constants (at -80 mV, SD, 3.8 ms, and FD, 0.13 ms). These two kinds of channels also differed in (a) activation threshold (SD, approximately -50 mV; FD, approximately -20 mV), (b) half-maximal activation (SD, between -15 and -10 mV; FD between +10 and +15 mV), and (c) inactivation time course (SD, fast; FD, slow). The total amplitude of the Ca current and the proportion of SD and FD channels varied from cell to cell. The amplitude of the K current was strongly dependent on the internal [Ca2+] and was almost abolished when internal [Ca2+] was less than 0.001 microM. The K current appeared to be independent, or only slightly dependent, of Ca influx. With an internal [Ca2+] of 0.1 microM, the activation threshold was -20 mV, and at +40 mV the half-time of activation was 9 ms. With 73 mM external K the closing time constant at -70 mV was approximately 3 ms. The outward current was also modulated by internal pH and Mg. At a constant pCa gamma a decrease of pH reduced the current amplitude, whereas the activation kinetics were not much altered. Removal of internal Mg produced a drastic decrease in the amplitude of the Ca-activated K current. It was also found that with internal [Ca2+] over 0.1 microM the K current underwent a time-dependent transformation characterized by a large increase in amplitude and in activation kinetics. 相似文献
16.
It is shown that the photoemf in a pigmented membrane is specific to the magnesium-porphyrin conductance channel. A null current method was devised to measure directly the voltage dependence of the photoemf and the magnesium-porphyrin conductance. Their voltage dependence is in agreement with the hypothesis that the magnesium-porphyrin cation is the majority carrier 相似文献
17.
Activation of voltage-dependent ion channels is primarily controlled by the applied potential difference across the membrane. For potassium channels the Drosophila Shaker channel has served as an archetype of all other potassium channels in studies of activation mechanisms. In the Shaker potassium channel much of the voltage sensitivity is conferred by the S4 transmembrane helix, which contains seven positively charged residues. During gating, the movement of these charges produces gating currents. Mutagenic and fluorescence studies indicate that four of these residues are particularly important and contribute to the majority of gating charge, R362, R365, R368 and R371. The channel is thought to dwell in several closed states prior to opening. Ionic-charge pairing with negatively charged residues in the S2 and S3 helices is thought to be important in regulating these closed states and detailed kinetic models have attempted to define the kinetics and charge of the transitions between these states. Neutral residues throughout the S4 and S5 helices are thought to control late steps in channel opening and may have important roles in modulating the stability of the open state and late closed states. In response to depolarization, the S4 helix is thought to undergo a rotational translation and this movement is also important in studies of the movement of the pore helices, S5 and S6, during opening. This review will examine residues that are important during activation as well as kinetic models that have attempted to quantitatively define the activation pathway of voltage-dependent potassium channels. 相似文献
18.
John S. Ramsdell 《In vitro cellular & developmental biology. Plant》1990,26(3):250-258
Summary GH cells are a widely used cell strain for the investigation of mechanisms regulating hormone release and synthesis. This
report identifies two inducible phenotypes of the GH4 clone (epithelioid and motile) which may extend studies of this well-characterized cell line to different stages of pituitary
cell development. GH4C1 cells treated in suspension with epidermal growth factor plus tetradecanoylphorbol acetate aggregate to form large epithelioid
colonies with extensive cell-to-cell and cell-to-substratum adhesion. These cells cease replicating within 48 h, increase
50% in cell volume, and synthesize 40-fold more prolactin. A GH4C1 variant with enhanced substratum adhesion and little or no cell-to-cell adhesion (GH4S1), responds differently to this treatment. These cells cease replicating immediately, show increased cell separation, develop
leading lamellae, and display locomotory activity. Each phenotype coexists in mixed cultures of GH4C1 and GH4S1 cells. This indicates that the different inducible response of the variant does not result from autocrine secretion. A molecular
basis for cell-to-cell adhesion in GH4 cells was investigated. GH4C1, but not the variant cells, express a 180 kDa immunoreactive protein indistinguishable from an isoform of the neural cell
adhesion molecule. Therefore the absence of cell-to-cell adhesion and inability to develop extensive cell-to-cell adhesion
characteristic of the epithelioid phenotype may result from altered expression of the neural cell adhesion molecule. These
findings are important because they have defined an in vitro approach to investigate genetic and cellular changes associated
with the development and progression of pituitary cell phenotype.
This study was initiated in the laboratory of Dr. A. H. Tashjian, Jr., under the support of grant DK11011 from the National
Institutes of Health, Bethesda, MD. The completion of this study was supported by the Medical University of South Carolina
Biomedical Support Grant of 1987–1988 and the American Cancer Society grant 1N-175. 相似文献
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Two voltage-dependent potassium channels, KCNQ1 and KCNH2, are expressed in the taste buds and were identified as strong candidates involved in the repolarization of taste receptor cells expressing phospholipase C-beta2 and TRPM5 (beta2/M5-TRCs). In cell type-specific expression, KCNQ1 was expressed in most taste bud cells, including beta2/M5-TRCs, whereas KCNH2 was expressed in a subset of beta2/M5-TRCs with no correlation with their taste modality, such as sweet or bitter taste reception. Expression of KCNH2 was restricted to young beta2/M5-TRCs. These results suggest that taste bud cells other than beta2/M5-TRCs are depolarized by some stimuli and also that beta2/M5-TRCs have cell age-dependent molecular mechanisms of repolarization. 相似文献