scholarly journals Characteristics of Five Types of K+ Channel in Cultured Locust Muscle

1990 ◽  
Vol 154 (1) ◽  
pp. 45-65
Author(s):  
P. N. Usherwood
Keyword(s):  
Single Channel ◽  
Inward Rectifier ◽  
Resting Potential ◽  
K Channel ◽  
Chord Conductance ◽  
Outward Currents ◽  
Inward Currents

K+ channel activity in cultured locust myofibres was investigated using gigaohm patch-clamp techniques. After 2 months in vitro the myofibres had a mean resting potential of −39 ± 7 mV (±S.D., 7V = 42). Five types of K+ channel were identified at this time. The majority of single-channel events recorded from cellattached patches were due to a small-conductance (type 1) and a largeconductance (type 2), inward rectifier, K+ channel. In cell-attached patches, with 180 mmoll−1 KC1 in the patch pipette, the type 1 channel had a chord conductance of 43 pS for inward currents and 8pS for outward currents; the type 2 channel had a chord conductance of 115 pS for inward currents and 29 pS for outward currents. The type 2 channel exhibited bursting kinetics, was ATP-sensitive and could be blocked by Ba2+. Two other channels (types 3 and 4) had linear conductances of 130pS and 207pS, respectively. The type 3 channel was Ca2+-sensitive. A further channel (type 5) appeared to be an inward rectifier with a conductance of 5pS. Openings of types 3, 4 and 5 channels occurred less frequently than openings of the other two channels. Types 1, 2, 3 and 4 channels possessed multiple closed and open states with non-linear gating mechanisms.

ATP-sensitive K(+)-selective channels of inner medullary collecting duct cells

1994 ◽  
Vol 267 (3) ◽  
pp. F489-F496 ◽  
Author(s):  
S. C. Sansom ◽  
T. Mougouris ◽  
S. Ono ◽  
T. D. DuBose
Keyword(s):  
Single Channel ◽  
Collecting Duct ◽  
K Channel ◽  
Channel Conductance ◽  
Single Channel Conductance ◽  
Inner Medullary Collecting Duct ◽  
Outward Currents ◽  
Inward Currents ◽  
Excised Patches ◽  
Medullary Collecting Duct

The inner medullary collecting duct (IMCD) in vivo has the capacity to either secrete or reabsorb K+. However, a selective K+ conductance has not been described previously in the IMCD. In the present study, the patch-clamp method was used to determine the presence and properties of K(+)-selective channels in the apical membrane of the inner medullary collecting duct cell line, mIMCD-3. Two types of K(+)-selective channels were observed in both cell-attached and excised patches. The most predominant K+ channel, a smaller conductance K+ channel (SK), was present in cell-attached patches with 140 mM KCl (high bath K+) but not with 135 mM NaCl plus 5 mM KCl (low bath K+) in the bathing solution. The single-channel conductance of SK was 36 pS with inward currents and 29 pS with outward currents in symmetrical 140 mM KCl. SK was insensitive to both voltage and Ca2+. However, SK was inhibited significantly by millimolar concentrations of ATP in excised patches. A second K(+)-selective channel [a larger K+ channel (BK)] displayed a single-channel conductance equal to 132 pS with inward currents and 90 pS with outward currents in symmetrical 140 mM KCl solutions. BK was intermittently activated in excised inside-out patches by Mg(2+)-ATP in concentrations from 1 to 5 mM. With complete removal of Mg2+, BK was insensitive to ATP. BK was also insensitive to potential and Ca2+ and was observed in cell-attached patches with 140 mM KCl in the bath solution. Both channels were blocked reversibly by 1 mM Ba2+ from the intracellular surface but not by external Ba2+.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Effect of Auxin (IAA) on the Fast Vacuolar (FV) Channels in Red Beet (Beta vulgaris L.) Taproot Vacuoles

2020 ◽  
Vol 21 (14) ◽  
pp. 4876
Author(s):  
Zbigniew Burdach ◽  
Agnieszka Siemieniuk ◽  
Waldemar Karcz
Keyword(s):  
Single Channel ◽  
Outward Current ◽  
K Channel ◽  
Single Channels ◽  
Patch Clamp Technique ◽  
Open Probability ◽  
Inward Current ◽  
Bath Solution ◽  
Outward Currents ◽  
Inward Currents

In contrast to the well-studied effect of auxin on the plasma membrane K+ channel activity, little is known about the role of this hormone in regulating the vacuolar K+ channels. Here, the patch-clamp technique was used to investigate the effect of auxin (IAA) on the fast-activating vacuolar (FV) channels. It was found that the macroscopic currents displayed instantaneous currents, which at the positive potentials were about three-fold greater compared to the one at the negative potentials. When auxin was added to the bath solution at a final concentration of 1 µM, it increased the outward currents by about 60%, but did not change the inward currents. The imposition of a ten-fold vacuole-to-cytosol KCl gradient stimulated the efflux of K+ from the vacuole into the cytosol and reduced the K+ current in the opposite direction. The addition of IAA to the bath solution with the 10/100 KCl gradient decreased the outward current and increased the inward current. Luminal auxin reduced both the outward and inward current by approximately 25% compared to the control. The single channel recordings demonstrated that cytosolic auxin changed the open probability of the FV channels at the positive voltages to a moderate extent, while it significantly increased the amplitudes of the single channel outward currents and the number of open channels. At the positive voltages, auxin did not change the unitary conductance of the single channels. We suggest that auxin regulates the activity of the fast-activating vacuolar (FV) channels, thereby causing changes of the K+ fluxes across the vacuolar membrane. This mechanism might serve to tightly adjust the volume of the vacuole during plant cell expansion.

scholarly journals Clotrimazole-sensitive K+ currents regulate pacemaker activity in interstitial cells of Cajal

2007 ◽  
Vol 292 (6) ◽  
pp. G1715-G1725 ◽  
Author(s):  
Yaohui Zhu ◽  
Jing Ye ◽  
Jan D. Huizinga
Keyword(s):  
Single Channel ◽  
K Channel ◽  
Pacemaker Activity ◽  
Short Term ◽  
Outward Currents ◽  
Inward Currents ◽  
Cells Of Cajal ◽  
K Currents ◽  
Short Term Culture

Interstitial cells of Cajal (ICC) are pacemaker cells for gut peristaltic motor activity. Compared with cardiac pacemaker cells, little is known about mechanisms that regulate ICC excitability. The objective of the present study was to investigate a potential role for clotrimazole (CTL)-sensitive K currents ( ICTL) in the regulation of ICC excitability and pacemaker activity. ICC were studied in situ and in short-term culture by using the whole cell patch-clamp configuration. In situ, ICC exhibited spontaneous transient inward currents followed by transient outward currents. CTL blocked outward currents, thereby increasing the net inward currents, and depolarized ICC, thereby establishing CTL-sensitive channels as regulators of ICC pacemaker activity. In short-term culture, a ICTL was identified that showed increased conductance when depolarized from the resting membrane potential to 0 mV and subsequent inward rectification at further depolarized potentials. The ICTL markedly increased with increasing intracellular calcium and was insensitive to the ether-à-go-go-related K channel blocker E-4031 and the large-conductance calcium-activated K channel blocker iberiotoxin. ICTL contributed 3–9 nS to the whole cell conductance at 0 mV membrane potential under physiological conditions; it was fast activating (τ = 88 ms), showed little time-dependent inactivation, and exhibited a deactivation time constant of 38 ms. The nitric oxide donor sodium nitroprusside (SNP) increased ICTL. Single-channel activity, activated by calcium and SNP, was inhibited by CTL, with a single-channel conductance of ∼38 pS. In summary, ICC generate a ICTL on depolarization through an intermediate-conductance calcium-activated K channel that regulates pacemaker activity and ICC excitability.

scholarly journals Effect of Different Fermentation Condition on Estimated Glycemic Index, In Vitro Starch Digestibility, and Textural and Sensory Properties of Sourdough Bread

Foods ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 514
Author(s):  
Hilal Demirkesen-Bicak ◽  
Muhammet Arici ◽  
Mustafa Yaman ◽  
Salih Karasu ◽  
Osman Sagdic
Keyword(s):  
Glycemic Index ◽  
Sensory Properties ◽  
Starch Digestibility ◽  
In Vitro Starch Digestibility ◽  
Whole Wheat ◽  
Sourdough Bread ◽  
Estimated Glycemic Index

This study aimed to evaluate the influence of sourdough fermentation on the estimated glycemic index (eGI), in vitro starch digestibility, and textural and sensory properties of eight experimentally prepared sourdough breads. Wheat and whole wheat flour bread samples were produced under different fermentation conditions (25 °C and 30 °C) and fermentation methods (type-1 and type-2). In type-1 fermentation, sourdough was obtained via spontaneous fermentation. Indigenous strains (Lactobacillus brevis ELB99, Lactiplantibacillus plantarum ELB75, and Saccharomyces cerevisiae TGM55) were used for type-2 fermentation. Fermentation type and temperature significantly affected eGI, the hydrolysis index (HI), the starch fraction, and the textural properties of the samples (p < 0.05). The resistant starch (RS) content increased after fermentation, while rapidly digestible starch (RDS), HI, and eGI decreased. RS values were significantly higher in type-2 than in type-1 at the same temperature for both flour types (p < 0.05). At 25 °C, RS values were higher in both fermentation types. In the white flour samples, eGI values were in the range of 60.8–78.94 and 62.10–78.94 for type-1 and type-2, respectively. The effect of fermentation type on eGI was insignificant (p < 0.05). In the whole flour samples, fermentation type and temperature significantly affected eGI (p < 0.05). The greatest eGI decreases were in whole wheat sourdough bread at 30 °C using type-2 (29.74%). The 30 °C and type-2 samples showed lower hardness and higher specific volume. This study suggests that fermentation type and temperature could affect the eGI and the textural and sensory properties of sourdough bread, and these factors should be considered during bread production. The findings also support the consumption of wheat and whole wheat breads produced by type-2 fermentation due to higher RS and slowly digestible starch (SDS) and lower RDS and eGI values.

scholarly journals First Report ofEurycoma longifoliaJack Root Extract Causing Relaxation of Aortic Rings in Rats

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Bae Huey Tee ◽  
See Ziau Hoe ◽  
Swee Hung Cheah ◽  
Sau Kuen Lam
Keyword(s):  
Erectile Function ◽  
Receptor Activation ◽  
Root Extract ◽  
Ang Ii ◽  
Angiotensin I ◽  
Eurycoma Longifolia ◽  
Vasodilatory Effect

AlthoughEurycoma longifoliahas been studied for erectile function, the blood pressure- (BP-) lowering effect has yet to be verified. Hence, this study aims at investigating the BP-lowering properties of the plant with a view to develop an antihypertensive agent that could also preserve erectile function. Ethanolic root extract was partitioned by hexane, dichloromethane (DCM), ethyl acetate, butanol, and water. The DCM fraction, found to be potent in relaxing phenylephrine- (PE-) precontracted rat aortic rings, was further purified by column chromatography. Subfraction DCM-II, being the most active in relaxing aortae, was studied for effects on the renin-angiotensin and kallikrein-kinin systems in aortic rings. The effect of DCM-II on angiotensin-converting enzyme (ACE) activity was also evaluatedin vitro. Results showed that DCM-II reduced (p<0.05) the contractions evoked by angiotensin I and angiotensin II (Ang II). In PE-precontracted rings treated with DCM-II, the Ang II-induced contraction was attenuated (p<0.05) while bradykinin- (BK-) induced relaxation enhanced (p<0.001).In vitro, DCM-II inhibited (p<0.001) the activity of ACE. These data demonstrate that the vasodilatory effect of DCM-II appears to be mediatedviainhibition of Ang II type 1 receptor and ACE as well as enhancement of Ang II type 2 receptor activation and BK activity.

scholarly journals Differential Activation of CD8+Tumor-Specific Tc1 and Tc2 Cells by an IL-10-Producing Murine Plasmacytoma

1998 ◽  
Vol 6 (3-4) ◽  
pp. 331-342 ◽  
Author(s):  
Christoph Specht ◽  
Hans-Gerd Pauels ◽  
Christian Becker ◽  
Eckehart Kölsch
Keyword(s):  
T Cells ◽  
Immune Responses ◽  
Tumor Cells ◽  
T Cell Subsets ◽  
Early Stages ◽  
Specific Tolerance

The involvement of counteractiveCD8+T-cell subsets during tumor-specific immune responses was analyzed in a syngeneic murine plasmacytoma model.CD8+Tc cells against the immunogenic IL-10-producing BALB/c plasmacytoma ADJ-PC-5 can be easily induced by immunization of BALB/c mice with X-irradiated ADJ-PC-5 tumor cellsin vivoandin vitro. However, the failure of recipient mice to mount a protective Tc response against the tumor during early stages of a real or simulated tumor growth is not due to immunological ignorance, but depends on the induction of tumor-specific tolerance, involving a population of tumorinducedCD8+T cells that are able to inhibit the generation of tumor-specific Tc cells in a primary ADJ-PC-5-specific MLTC, using IFN-γas a suppressive factor. Whereas most longterm cultivated CD8+ADJ-PC-5-specific Tc lines produce type-1 cytokines on stimulation, at least two of them, which were derived from a primary MLTC, display a type-2 cytokine spectrum. Furthermore, the primaryin vitroTc response against ADJ-PC-5 cells shows characteristics of a Tc2 response. The Tc response is strictly depending on tumor-derived IL-10.CD8+Tc cells that are induced in a primary MLTC do not produce IFN-γ, and the tumor-specific Tc response is enhanced by IL-4 but suppressed by IFN-γor IL-12. In contrast, ADJ-PC- 5-specificCD8+Tc cells from immunized mice are IFN-γproducing Tc1 cells. Since the primaryin vitroTc response against the tumor is suppressed even by the smallest numbers of irradiated ADJ-PC-5-specific Tc1 cells via IFN-γthese Tc1 cells behave similar to the suppressiveCD8+T cells that are induced during early stages of ADJ-PC-5 tumorigenesis.

Permeation and gating properties of a cloned renal K+ channel

1995 ◽  
Vol 268 (2) ◽  
pp. C389-C401 ◽  
Author(s):  
S. Chepilko ◽  
H. Zhou ◽  
H. Sackin ◽  
L. G. Palmer
Keyword(s):  
Single Channel ◽  
Reversal Potential ◽  
Cation Binding ◽  
K Channel ◽  
Patch Clamp Technique ◽  
Open Probability ◽  
Membrane Hyperpolarization ◽  
Channel Current ◽  
Inward Currents ◽  
Kinetics Of

The renal K+ channel (ROMK2) was expressed in Xenopus oocytes, and the patch-clamp technique was used to assess its conducting and gating properties. In cell-attached patches with 110 mM K+ in the bath and pipette, the reversal potential was near zero and the inward conductance (36 pS) was larger than the outward conductance (17 pS). In excised inside-out patches the channels showed rectification in the presence of 5 mM Mg2+ on the cytoplasmic side but not in Mg(2+)-free solution. Inward currents were also observed when K+ was replaced in the pipette by Rb+, NH4+, or thallium (Tl+). The reversal potentials under these conditions yielded a selectivity sequence of Tl+ > K+ > Rb+ > NH4+. On the other hand, the slope conductances for inward current gave a selectivity sequence of K+ = NH4+ > Tl+ > Rb+. The differences in the two sequences can be explained by the presence of cation binding sites within the channel, which interact with Rb+ and Tl+ more strongly and with NH4+ less strongly than with K+. Two other ions, Ba2+ and Cs+, blocked the channel from the outside. The effect of Ba2+ (1 mM) was to reduce the open probability of the channels, whereas Cs+ (10 mM) reduced the apparent single-channel current. The effects of both blockers are enhanced by membrane hyperpolarization. The kinetics of the channel were also studied in cell-attached patches. With K+ in the pipette the distribution of open times could be described by a single exponential (tau 0 = 25 ms), whereas two exponentials (tau 1 = 1 ms, tau 2 = 30 ms) were required to describe the closed-time distribution. Hyperpolarization of the oocyte membrane decreased the open probability and tau 0, and increased tau 1, tau 2, and the number of long closures. The presence of Tl+ in the pipette significantly altered the kinetics, reducing tau 0 and eliminating the long-lived closures. These results suggest that the gating of the channel may depend on the nature of the ion in the pore.

Ion channels in spinal cord astrocytes in vitro. I. Transient expression of high levels of Na+ and K+ channels

1992 ◽  
Vol 68 (4) ◽  
pp. 985-1000 ◽  
Author(s):  
H. Sontheimer ◽  
J. A. Black ◽  
B. R. Ransom ◽  
S. G. Waxman
Keyword(s):  
Spinal Cord ◽  
Membrane Potentials ◽  
Resting Potential ◽  
K Channel ◽  
Na Channels ◽  
Patch Clamp Recording ◽  
K Channels ◽  
Na Currents ◽  
Channel Expression

1. Na+ and K+ channel expression was studied in cultured astrocytes derived from P--0 rat spinal cord using whole cell patch-clamp recording techniques. Two subtypes of astrocytes, pancake and stellate, were differentiated morphologically. Both astrocyte types showed Na+ channels and up to three forms of K+ channels at certain stages of in vitro development. 2. Both astrocyte types showed pronounced K+ currents immediately after plating. Stellate but not pancake astrocytes additionally showed tetrodotoxin (TTX)-sensitive inward Na+ currents, which displayed properties similar to neuronal Na+ currents. 3. Within 4-5 days in vitro (DIV), pancake astrocytes lost K(+)-current expression almost completely, but acquired Na+ currents in high densities (estimated channel density approximately 2-8 channels/microns2). Na+ channel expression in these astrocytes is approximately 10- to 100-fold higher than previously reported for glial cells. Concomitant with the loss of K+ channels, pancake astrocytes showed significantly depolarized membrane potentials (-28.1 +/- 15.4 mV, mean +/- SD), compared with stellate astrocytes (-62.5 +/- 11.9 mV, mean +/- SD). 4. Pancake astrocytes were capable of generating action-potential (AP)-like responses under current clamp, when clamp potential was more negative than resting potential. Both depolarizing and hyperpolarizing current injections elicited overshooting responses, provided that cells were current clamped to membrane potentials more negative than -70 mV. Anode-break spikes were evoked by large hyperpolarizations (less than -150 mV). AP-like responses in these hyperpolarized astrocytes showed a time course similar to neuronal APs under conditions of low K+ conductance. 5. In stellate astrocytes, AP-like responses were not observed, because the K+ conductance always exceeded Na+ conductance by at least a factor of 3. Thus stellate spinal cord astrocyte membranes are stabilized close to EK as previously reported for hippocampal astrocytes. 6. It is concluded that spinal cord pancake astrocytes are capable of synthesizing Na+ channels at densities that can, under some conditions, support electrogenesis. In vivo, however, AP-like responses are unlikely to occur because the cells' resting potential is too depolarized to allow current activation. Thus the absence of electrogenesis in astrocytes may be explained by two mechanisms: 1) a low Na-to-K conductance ratio, as in stellate spinal cord astrocytes and in other previously studied astrocyte preparations; or, 2) as described in detail in the companion paper, a mismatch between the h infinity curve and resting potential, which results in Na+ current inactivation in spinal cord pancake astrocytes.

Sign in / Sign up

Export Citation Format

Share Document