Biomedical subjects

T Tokimasa

Publications and source records attributed to T Tokimasa.

At least 19 recordsLinked to original sources

PubMed2001

Hyperpolarizing shift by quinine in the steady-state inactivation curve of delayed rectifier-type potassium current in bullfrog sympathetic neurons.

Whole-cell recordings were made from dissociated bullfrog sympathetic neurons to examine the actions of quinine (1-100 microM) on the steady-state activation and inactivation curves of a delayed rectifier-type potassium current (I(K)). Quinine (EC50 approximately 8 microM) caused a hyperpolarizing shift (approximately 31 mV with 30 microM) in the inactivation curve of I(K) without significantly affecting its activation curve. Quinine (20 microM) was without effects on the voltage-dependence of a rapidly-inactivating A-type potassium current (I(A)). It is concluded that quinine can selectively modulate the voltage-dependence of I(K) in amphibian autonomic neurons.

Animals↗

PubMed2000

Effects of lanthanides on voltage-dependent potassium currents in bullfrog sympathetic neurons.

The effects of lanthanides (La(3+), Gd(3+), Lu(3+) and Sm(3+)) on voltage-dependent potassium currents were studied in dissociated bullfrog sympathetic neurons. A-type current (I(A)) and M-type current (I(M)) were blocked by lanthanides (0.1-30 microM) with I(M) being much less sensitive to these ions than I(A). The order of potency was Gd(3+)>/=Lu(3+) approximately La(3+) approximately Sm(3+) for I(A) and Gd(3+)&z.Gt;Lu(3+) approximately La(3+)>Sm(3+) for I(M). The I(M) block occurred independently of its activation kinetics while the I(A) block was associated with a positive shift of the activation and inactivation curves. Gd(3+) (100 microM) blocked the delayed rectifier-type current (I(K)) by less than 20%; Lu(3+), La(3+) and Sm(3+) (100 microM for each) were without effect on I(K). It is concluded that I(A) was the most sensitive to lanthanides, and Gd(3+) was the most potent for all the currents in amphibian autonomic neurons.

Animals↗

PubMed2000

Mechanisms underlying the M-current block by barium in bullfrog sympathetic neurons.

Whole-cell/voltage-clamp recordings were made from dissociated bullfrog sympathetic neurons to examine the channel blocking actions of barium (3-2000 microM) on an M-type potassium current (I(M)). Barium (IC(50) approximately 105 microM) blocked I(M) without affecting the 50%-activation voltage ( approximately -35 mV) and the slope factor ( approximately 11 mV) of the activation curve. The results indicate that the barium block is independent of the kinetics of I(M).

Action Potentials↗

PubMed1999

Effects of quinine on three different types of potassium currents in bullfrog sympathetic neurons.

Whole-cell/voltage-clamp recordings were made from dissociated bullfrog sympathetic neurons to examine the sensitivity of potassium currents to a potassium channel blocker quinine (1-500 microM). Among three currents tested, a rapidly inactivating A-type current (I(A)) was the most sensitive to the block by quinine (IC50 approximately 22 microM). A non-inactivating M-type current (I(M)) was the least sensitive (IC50 approximately 445 microM), and the sensitivity of a slowly inactivating delayed rectifier-type current (I(K)) was in between (IC50 approximately 115 microM). Results suggest that the ability of quinine to block different types of potassium currents such as I(A) and I(M) with significantly different IC50 values would be of help for the potassium channel pharmacology in amphibian autonomic ganglion cells.

Animals↗

PubMed1999

Theoretical background for inward rectification.

Theoretical background has been reviewed for inward rectification due to a potassium current termed IRK. The Eyring rate theory in which the thermodynamic rate coefficient for chemical reactants (channels and ions in this case) can be described in terms of energy barriers for potassium ions can mimic not only the polarity and degree of rectification but also the voltage-dependence of the barium-induced IRK block. The model predicts that the blocking site locates 30-70% depth from the outer margin of the IRK channel.

Animals↗

PubMed1998

Actions of zinc on rapidly inactivating A-type and non-inactivating M-type potassium currents in bullfrog sympathetic neurons.

The actions of zinc on A-type potassium current (I(A)) were studied in dissociated bullfrog sympathetic neurons. Zinc (1-300 microM) caused a parallel shift in the activation and inactivation curves to a depolarizing direction, thereby enhancing I(A) around physiological resting potential. An EC50 value was 70-100 microM for these actions. The zinc actions were non-selective in a sense that zinc inhibited M-type potassium current (I(M)) with an IC50 value of 300 microM. Zinc was without effect on the maximum conductance for I(A) and the kinetic behavior for I(M). The ability of low concentrations of zinc to modulate separate set of potassium currents such as I(A) and I(M) in conceptually distinct manner may therefore assume pathophysiological importance for autonomic neurons.

Animals↗

PubMed1998

Effects of barium on delayed rectifier potassium current in bullfrog sympathetic neurons pretreated with wortmannin.

The effect of barium (1 mM) on a delayed rectifier-type potassium current was examined in bullfrog sympathetic neurons. An M-type potassium current was eliminated by pretreatment of the cells with a microbial product, wortmannin (10 microM). An A-type potassium current was continuously inactivated by setting a holding potential at -65 mV. In treated cells (n = 10), the delayed rectifier at 0 mV averaged 2200 +/- 107 pA in the presence of barium (1 mM) as compared to 2308 +/- 110 pA in the controls, and 2085 +/- 103 pA after washing out the barium. It is concluded that the delayed rectifier is insensitive to barium blockage in amphibian autonomic neurons.

Androstadienes↗

PubMed1997

Evidence for the calcium-dependent potentiation of M-current obtained by the ratiometric measurement of the fura-2 fluorescence in bullfrog sympathetic neurons.

Intracellular Ca2+ concentration ([Ca]i) was measured following the activation of an inward Ca2+ current and subsequent potentiation of an M-type K+ current (IM) in bullfrog sympathetic neurons. Fura-2 was used as an indicator for [Ca]i. The fluorescence ratio at 340 and 380 nm (F340/F380) was elevated from 0.36 to 1.22 when IM was potentiated by 68% following the Ca2+ current. Based on the in vivo calibration curve obtained from cells permeabilized with digitonin (20 microM), the F340/F380 value of 1.22 was equivalent to a [Ca]i of 0.97 microM. We therefore propose that a rise in [Ca]i into the micromolar range can lead to the potentiation of IM in amphibian autonomic neurons.

Action Potentials↗

PubMed1997

Actions of barium on rapidly inactivating potassium current in bullfrog sympathetic neurons.

Whole-cell/voltage-clamp recordings were made from dissociated bullfrog sympathetic neurons to examine the inhibitory actions of barium (0.01-3 mM) on a rapidly inactivating A-type potassium current (IA). The IC50 value was about 0.9 mM. Barium (1 mM) approximately halved the maximum amplitude of IA (approximately 1.7 nA near 0 mV) without significantly affecting a voltage for the 50%-activation (approximately -40 mV) and that for the 50%-inactivation (approximately -90 mV), nor did it affected the time course of IA. The results suggest that the barium block is independent of the kinetics of the A-channels in bullfrog sympathetic neurons.

Animals↗

PubMed1997

M-type potassium current in dissociated sympathetic ganglion cells of Xenopus laevis.

Whole-cell/voltage-clamp recordings were made from dissociated sympathetic neurons of Xenopus laevis. Step depolarization (10-70 mV, 0.5 s) from the holding potential of -65 mV activated a non-inactivating potassium current which was selectively blocked by barium (1 mM). The current did not require an inward calcium current for its activation. The steady-state activation curve of the current was centered at -35 mV with a slope factor of 8.6 mV. The time constant for activation of the current was about 115 ms when measured at -35 mV and the effective valence of the voltage-sensing particle was 2.9. This potassium current was identified as a particular set of potassium currents usually referred to as M-current.

Animals↗

PubMed1996

Effects of barium, lanthanum and gadolinium on endogenous chloride and potassium currents in Xenopus oocytes.

1. The effects of multivalent cations on membrane currents recorded from Xenopus oocytes were studied. 2. The hyperpolarization-activated chloride current was reversibly blocked by lanthanum; half-maximal block occurred at a concentration of 8 microM. Zinc, cadmium, cobalt and nickel were less potent than lanthanum, and gadolinium, manganese, barium and strontium had no effect at a concentration of 100 microM. 3. The calcium-activated chloride current was blocked by gadolinium (50 microM), and lanthanum, cadmium, cobalt, nickel and manganese were equally effective. The actions of gadolinium and lanthanum were almost irreversible, while partial (30-80%) recovery was observed with the other cations. Zinc (100 microM) had no effect. 4. In lanthanum (100 microM), membrane depolarizations from -70 mV activated an outward potassium current that was partially blocked by barium (0.1-2 mM). The barium-sensitive current was confined to potentials less negative than -70 mV. The current consisted of a time-independent as well as a time-dependent component, the latter of which had voltage dependence similar to the M-current. 5. It is proposed that lanthanum, gadolinium and barium can usefully separate these endogenous membrane currents in Xenopus oocytes.

Animals↗

PubMed1996

Calcium-dependent after-hyperpolarization in dissociated bullfrog sympathetic neurons.

Whole-cell recordings were made from dissociated bullfrog sympathetic neurons. Tetraethylammonium (30 mM) and apamin (100 nM) were added to the superfusate to eliminate the known calcium-activated potassium currents termed Ic and IAHP. Under these conditions, the action potential carried by calcium ions was followed by a prolonged (10-60 s) after-hyperpolarization. A current component (IAC) underlying the after-hyperpolarization was eliminated by barium (2 mM) and showed voltage-dependence identical to that of a M-type potassium current. I concluded that the after-hyperpolarization is caused not only by IAHP but also by the calcium-dependent potentiation of M-current.

Animals↗

PubMed1996

Calcium-dependent potentiation of M-current in bullfrog sympathetic neurons.

Whole-cell voltage-clamp recordings were made from cultured bullfrog sympathetic neurons to measure the steady-state activation curve of M-type potassium current. When measured with a calcium-deficient (10 nM) pipette solution M-conductance was 4.8 nS at -35 mV having the 50%-activation voltage at-20 mV. Respective values were 17.2 nS at -35 mV with the 50%-activation voltage at -42 mV when measured with a calcium-rich (1 microM) solution, indicating the hyperpolarizing displacement of the activation curve with high internal calcium. It is suggested that intracellular calcium ions can modulate kinetics of M-current which thereby regulate the number of M-channels being open at given membrane potentials.

Animals↗

PubMed1996

Purinergic cation channels in neurons of rabbit vesical parasympathetic ganglia.

Membrane current was recorded from neurons in rabbit vesical parasympathetic ganglia, utilizing single electrode voltage clamp techniques. ATP (0.1-1 mM) caused an inward current (IATP) associated with an increased conductance at a holding potential of -50 mV. ADP (0.1-1 mM) and 5'-O-3-thiotriphosphate (0.1-0.6 mM) but not AMP (0.3-2 mM) and adenosine (0.1-2 mM) mimicked the actions of ATP. The IATP reversed its polarity at -12.1 +/- 1.4 mV. The amplitude of the IATP was depressed in low sodium solutions and in nominally calcium-free solutions but not in low chloride solutions. Suramin (10-100 microM) and reactive blue 2 (10-100 microM), P2-antagonists, reversibly depressed the IATP. In contrast, hexamethonium (100 microM) did not affect the IATP. These data suggest that ATP activates cation channels through P2X receptor subtypes in parasympathetic neurons.

Animals↗

PubMed1996

Hyperpolarizing shift of the M-current activation curve after washout of muscarine in bullfrog sympathetic neurons.

The mechanism underlying the over-recovery of an M-type potassium current following the washout of muscarine (20 microM) has been examined. Whole-cell recordings were made from single neurons dissociated from bullfrog sympathetic ganglia. During over-recovery, the maximum M-conductance decreased by about 2.8 nS while the steady-state M-current activation curve was displaced in the hyperpolarizing direction by about 13 mV. These data suggest that a hyperpolarizing shift in the kinetics of M-current causes over-recovery in amphibian autonomic neurons.

Animals↗

PubMed1996

Electrophysiological studies of the cell membrane of bullfrog sympathetic neurons.

Active and passive properties of dissociated bullfrog sympathetic ganglion cells were examined in the whole-cell configuration. Electrical constants were 107 k omega cm2 and 1 microFcm2 for the unitary cell membrane area. Under conditions in which the equilibrium potential was 55 mV for sodium ions and -101 mV for potassium ions, the action potential and its after-hyperpolarization peaked at 50 and -83 mV, respectively, when elicited from the resting potential of -63 mV. These observations are consistent with those reported previously for other autonomic neurons. The reversal potential of the after-hyperpolarization was approximately 10 mV less negative than the potassium equilibrium potential. The basis of this difference has been discussed using a hypothetical equivalent circuit.

Action Potentials↗

PubMed1995

Effects of myosin light chain kinase inhibitors on delayed rectifier potassium current in bullfrog sympathetic neurons.

Actions of myosin light chain kinase inhibitors were tested on delayed rectifier potassium current (IK) in dissociated bullfrog sympathetic neurons. A microbial product, wortmannin (10 microM, extracellularly) and a synthetic peptide, SM-1 (20 microM, intracellularly) caused approximately 35 mV hyperpolarizing shift of the inactivation curve. Substitution of ATP (1.15 mM) in the pipette solution with 5'-adenylylimidodiphosphate mimicked the actions of wortmannin and SM-1. Results suggest that phosphorylation of myosin may modulate kinetics for the inactivation of IK.

Adenylyl Imidodiphosphate↗

PubMed1995

Inhibition by wortmannin of M-current in bullfrog sympathetic neurones.

1. The actions of wortmannin, an inhibitor of myosin light chain kinase (MLCK), on M-type potassium current of dissociated bullfrog sympathetic neurones have been examined. 2. The amplitude of M-current was measured by whole cell recordings from cells pretreated with wortmannin (0.01-10 microM) or the wortmannin vehicle, dimethylsulphoxide (0.0001-0.1 vol%), for 30 min. Internal (recording pipette) solutions having three different pCa values (6, 7 and 8) were used for the measurements. 3. Irrespective of the pCa, M-current was not detectable when the cells were pretreated with 10 microM wortmannin. Wortmannin, 3 microM, produced 85-95% inhibition of the M-current. Pretreatment with 10-30 nM wortmannin was without effect on M-current. 4. The M-current inhibition by wortmannin at concentrations of 0.1-1 microM depended on the pCa of the internal solution. Inhibition occurred only when the calcium-rich (pCa = 6) internal solution was used. 5. Pre-treatment of the cells with wortmannin (10 microM) did not affect rapidly-inactivating A-type or delayed rectifier-type potassium currents not did it alter inwardly rectifying sodium-potassium current (IH). 6. These observations show that M-current inhibition by wortmannin has two pharmacological profiles. One is calcium-dependent and occurs at lower concentrations (0.1-1 microM), and is attributed to inhibition of MLCK by wortmannin. At higher concentrations (3-10 microM), wortmannin has an additional, calcium-independent action, inhibiting the M-current by an unknown mechanism.

Androstadienes↗