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I Findlay

Publications and source records attributed to I Findlay.

At least 73 records · Page 4Linked to original sources
PubMed1988

Calcium-dependent inactivation of the ATP-sensitive K+ channel of rat ventricular myocytes.

Single-channel currents were recorded from ATP-sensitive K+ channels in inside-out membrane patches excised from isolated rat ventricular myocytes. Perfusion of the internal surface of excised membrane patches with solutions which contained between 5 and 100 microM free calcium caused the loss of K+ATP channel activity which was not reversed when the membranes were washed with Ca-free solution. K+ATP channel activity could be recovered by bathing the patches in Mg.ATP. The loss of K+ATP channel activity provoked by internal calcium was a process which occurred over a time scale of seconds. Channel closure evoked by internal ATP was essentially instantaneous. The speed of K+ATP channel inactivation increased with the concentration of calcium. Neither a phosphatase inhibitor (fluoride ions) nor a proteinase inhibitor (leupeptin) had any effect upon the loss of K+ channel activity stimulated by internal calcium.

Adenosine Triphosphate↗
PubMed1988

ATP4- and ATP.Mg inhibit the ATP-sensitive K+ channel of rat ventricular myocytes.

K+ currents were recorded from ATP-sensitive channels in inside-out membrane patches excised from isolated rat ventricular myocytes. ATP-sensitive K+ channel inhibition could be evoked by ATP in the absence of magnesium where most ATP would be present as the free acid ATP4-. Channel inhibition was enhanced when the same total concentration of ATP was applied in the presence of magnesium, where most ATP would be bound as ATP.Mg. Dose-response relationships for ATP-sensitive K+ channel inhibition evoked by ATP had a Hill coefficient of 2 and Ki of 17 and 30 microM for ATP in the presence and absence of magnesium respectively. This was the obverse of the expected results if ATP4- were to be the sole form of ATP to effect channel closure. ATP-sensitive K+ channel inhibition evoked by ATP gamma S, AMP-PNP and AMP-PCP was also enhanced in the presence of magnesium. It is concluded that the ATP-sensitive K+ channel of rat ventricular myocytes binds and is closed by both the free-acid and divalent-cation-bound forms of ATP.

Adenosine Triphosphate↗
PubMed1988

Effects of ADP upon the ATP-sensitive K+ channel in rat ventricular myocytes.

The effects of ADP upon the gating of ATP-sensitive K+ channels from rat ventricular myocytes have been investigated by patch-clamp single-channel current recording experiments. ADP was applied to the internal surface of excised inside-out membrane patches and depending upon the experimental protocol and the concentration it was found that ADP could either inhibit or stimulate openings of ATP-sensitive K+ channels. In the absence of inactivation, ATP-sensitive K+ channels were inhibited by ADP in a dose-dependent manner. Partially inactivated channels, on the other hand, were stimulated by low (10 to 250 microM) and inhibited by high (greater than 250 microM) concentrations of ADP. ATP-sensitive K+ channels which were being inhibited by ATP (less than 1 mM) could be opened by the simultaneous application of ADP (50 microM to 1 mM). ADP had no effect upon channels inhibited by mM concentrations of ATP. The situation was further complicated when it was found that inhibition evoked by ADP was strongly attenuated by the presence of Mg2+ ions whilst channel stimulation, whether of partially inactivated channels or channels inhibited by ATP, required the presence of Mg2+ ions. The analog of ADP, ADP beta S, always evoked inhibition of ATP-sensitive K+ channels which was not affected by the presence or absence of Mg2+ ions.

Adenosine Diphosphate↗
PubMed1988

Effects of pyridine nucleotides on the gating of ATP-sensitive potassium channels in insulin-secreting cells.

The single-channel current recording technique has been used to study the influences that the pyridine nucleotides NAD, NADH, NADP and NADPH have on the gating of ATP-sensitive K+ channels in an insulin-secreting cell line (RINm5F). The effects of the nucleotides were studied at the intracellular surface using either excised inside-out membrane patches or permeabilized cells. All four pyridine nucleotides were found to evoke similar effects. At low concentrations, 100 microM and less, each promoted channel opening whereas high concentrations, 500 microM and above, evoked channel closure. The degree of K+ channel activation by pyridine nucleotides (low conc.) was found to be similar to that evoked by the same concentrations of ADP or GTP, whereas the degree of K+ channel inhibition (high conc.) was less marked than that evoked by the same concentrations of ATP, and never resulted in refreshment of K+ channels following removal. The effects of NAD, NADH, NADP and NADPH seemed to interact with those of ATP and ADP. In the presence of 1 mM ADP and 4 mM ATP, 10 to 100 microM concentrations of the pyridine nucleotides could not evoke channel opening, whereas concentrations of 500 microM and above were found to evoke channel closure. In the presence of 2 mM ATP and 0.5 mM ADP, however, 10 to 100 microM concentrations of the pyridine nucleotides were able to activate K+ channels.

Adenosine Diphosphate↗
PubMed1988

Effect of marathon training on the plasma lactate response to submaximal exercise in middle-aged men.

Twenty-one previously sedentary male volunteers (aged 35-50 years) undertook a defined marathon training programme lasting 30 weeks. At weeks 0 (T1), 15 (T2) and 30 (T3) they underwent measurement of maximal oxygen uptake (VO2 max), submaximal VO2 and submaximal plasma lactate concentration during cycle ergometry. No exercise was taken for 24-48 hours prior to testing. During training aerobic power increased significantly (p less than 0.001) from an initial VO2 max at T1 of 33.9 +/- 6 (mean +/- sd) ml.kg-1min-1 to 39 +/- 5.6 ml.kg-1min-1 at T2 but the T3 value of 39.2 +/- 5.2 ml.kg-1min-1 was not significantly different from that at T2. Plasma lactate concentration of 4 mmol.l-1 (OBLAw) occurred at a significantly (P less than 0.05) higher workload (155 +/- 28 w) at T2 compared with T1 (132 +/- 30 w) but the T3 figure was 137 +/- 34 w. OBLA VO2 at T1 was 2.04 +/- 0.42 l.min-1, at T2 was 2.24 +/- 0.04 l.min-1 but at T3 was 2.03 +/- 0.30 l.min-1 (T1:T2 P less than 0.05, T1:T3 NS). OBLA % VO2 max at T1 was 75 +/- 12%, at T2 was 73 +/- 11% but at T3 was 62 +/- 10% (T1:T2 NS, T1:T3 P less than 0.01).

Adult↗
PubMed1987

ATP-sensitive K+ channels in rat ventricular myocytes are blocked and inactivated by internal divalent cations.

K+ currents were recorded from ATP-sensitive channels in inside-out patches from isolated rat ventricular myocytes. In the absence of internal divalent cations the current voltage relationship could be described by constant-field assumptions with a permeability of 1.25 X 10(-13) cm2/s; outward currents saturated under a high driving force for K+ movement. Internal 0.1-5.0 mM Mg2+, 0.1 microM Ca2+ and 10 mM Na+ each depressed the flux of K+ ions moving outwards through open channels. Internal 0.1-5.0 mM Mg2+, 0.1-1.0 microM Ca2+ and 1-10 microM Ba2+ and Sr2+ blocked K+ channel activity in a dose- and voltage-dependent manner. Run-down channels could be reactivated by Mg-ATP, but not by AMP-PNP, ATP gamma S or Mg-free ATP which suggested that phosphorylation of the channels was involved in their activity. Ca2+ (greater than = 1 microM) and Sr2+ (1 mM) markedly inactivated K+ ATP channels, millimolar Ba2+ or Mg2+ were less effective. This suggested that the run down of the channels was a Ca2+-dependent dephosphorylation of the K+ channel protein.

Adenosine Triphosphate↗
PubMed1987

The effects of magnesium upon adenosine triphosphate-sensitive potassium channels in a rat insulin-secreting cell line.

1. The patch-clamp method of single-channel recording was applied to K+ channels which are inhibited by intracellular adenosine 5'-triphosphate (ATP: K+-ATP channels) in membrane patches obtained from the insulin-secreting cloned cell line RINm5F. 2. The magnitude of K+ currents flowing outwards through these K+-ATP channels was reduced by internal Mg2+ ions in a dose-dependent manner. Currents flowing inwards through the channels were not affected by Mg2+. Internal Na+ ions had similar effects. 3. Divalent cations (Mg2+, Sr2+ and Ca2+) applied to the internal surface of the patch membrane inhibited the opening of K+-ATP channels in a dose-dependent manner. Internal Na+ ions had no effect. 4. K+-ATP channel activity was stimulated by adenosine 5'-diphosphate (ADP), guanosine 5'-triphosphate (GTP), guanosine 5'-diphosphate (GDP), guanosine 5'-o-(3-thiotriphosphate) (GTP gamma S) and guanosine 5'-o-(2-thiodiphosphate) (GDP beta S) when millimolar Mg2+ bathed the internal surface of the patch membrane. In the virtual absence of internal Mg2+ ions ADP, GTP, and GTP gamma S inhibited K+-ATP channels and GDP and GDP beta S were without effect. Adenosine 5'-o-(2-thiodiphosphate) (ADP beta S) inhibited K+-ATP channel activity in the presence and absence of Mg2+. 5. K+-ATP channel openings could be evoked by either ADP or GDP in the presence of an inhibitory concentration of ATP. These openings were abolished in the absence of internal Mg2+. 6. Run-down K+-ATP channels could be reactivated by ATP in the presence of internal Mg2+, but not in its absence. Analogues of ATP were unable to reactivate K+-ATP channels even in the presence of Mg2+. 7. It is concluded that internal Mg2+ ions (i) cause the rectification of the K+-ATP channel current-voltage relationship, (ii) are required for K+-ATP channel activity to be maintained by a phosphorylation process and (iii) are required for K+-ATP channel activity evoked by ADP, GTP and GDP.

Action Potentials↗
PubMed1986

Combination of verapamil and beta blockers in systemic hypertension.

The efficacy and safety of verapamil and propranolol were examined in 14 hypertensive patients (mean age 51.2, range 30 to 65) in a double-blind, randomized, crossover study of verapamil, 360 mg, propranolol, 240 mg, these 2 formulations in combination and placebo, each given for 4 weeks. Supine blood pressure, heart rate, atrioventricular conduction (PR interval) and left ventricular function were measured. All treatments reduced diastolic blood pressure (mean +/- standard deviation) (p less than 0.001): placebo to 106.6 +/- 8.1 mm Hg; propranolol to 93.8 +/- 7.7; verapamil to 89.8 +/- 7.8; the combination to 84.1 +/- 6.1, but the effect of the combination was significantly greater than that of either drug alone (p less than 0.05). Heart rate at rest (placebo, 80.2 +/- 12.2 beats/min) was reduced by propranolol (63.3 +/- 9.4, p less than 0.001), but not by verapamil (79.0 +/- 8.9). However, the addition of verapamil to propranolol led to a further reduction in heart rate (56.9 +/- 8.4, p less than 0.005). PR interval was prolonged significantly by the combination (185.5 +/- 35.3 ms) when compared with placebo (154.0 +/- 22.7); propranolol (159.1 +/- 21.2) and verapamil (165.5 +/- 32.4) (p less than 0.005 for each). The active drugs increased end-diastolic dimension and end-systolic dimension. For each variable, the effect of the combination was statistically significant (p less than 0.01). Fractional shortening was not altered significantly by any of the treatments. Thus verapamil plus propranolol is a very effective antihypertensive combination but heart rate, atrioventricular conduction and left ventricular function may be affected adversely, necessitating careful monitoring of therapy.

Adult↗
PubMed1986

ATP maintains ATP-inhibited K+ channels in an operational state.

In patch-clamp records of K+ ATP channels in an insulin-secreting cell line (RINm5F) inhibition evoked by exposing the internal surface of the membrane to ATP is followed not just by the recovery of K+ ATP channel activity when the ATP is removed but by a marked activation of K+ ATP channels. This phenomenon is not a direct consequence of channel closure as inhibition induced by quinidine and quinine is followed upon the removal of the drug only by the recovery of K+ ATP channel activity and not by post-inhibitory activation. If ATP is applied to the exposed internal surface of a membrane patch when all of its K+ ATP channel have run down subsequent removal of the ATP causes their activation. The magnitude and duration of the reactivation of K+ ATP channels is shown to depend upon both the concentration of ATP and the length of time for which the membrane is exposed to ATP. We therefore have a paradoxical situation in that K+ channels which are inhibited by intracellular ATP require intracellular ATP to retain the ability to open.

Adenosine Triphosphate↗
PubMed1986

ATP-sensitive K+ channels in an insulin-secreting cell line are inhibited by D-glyceraldehyde and activated by membrane permeabilization.

The control of K+ channels in the insulin-secreting cell line RINm5F has been investigated by patch-clamp single-channel current recording experiments. The unitary current events recorded from cell-attached patches are due to large and small inwardly rectifying ATP-sensitive K+ channels with conductance properties similar to the two channels previously identified in primary cultured rat islet cells (Findlay, I., Dunne, M.J., & Petersen, O.H. J. Membrane Biol. 88:165-172, 1985). Cell permeabilization through brief exposure to 10 microM digitonin or 0.05% saponin (outside the isolated membrane patch area) results in a dramatic increase in current through the cell-attached patch due to opening of many large and small K+-selective channels. These channels are inhibited in a dose-dependent manner by ATP applied to the bath (near-complete inhibition by 5 mM ATP). During prolonged ATP exposure (1-5 min) the initial inhibition is followed by partial recovery of channel activity, although further activation does occur when ATP is subsequently removed. From the maximal number of coincident channel openings in the permeabilized cells (in the absence of ATP), it is estimated that there are on average 12 large ATP-sensitive K+ channels per membrane patch, but in the intact cells less than 5% of the membrane patches exhibited three or more coincident K+ channel openings, indicating the degree to which the channels are inhibited in the resting condition by endogenous ATP. Stimulation of RINm5F cells to secrete insulin was carried out by challenging intact cells with 10 mM D-glyceraldehyde.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1986

Messenger-mediated control of potassium channels in secretory cells.

In exocrine acinar cells (pancreas, salivary gland, lacrimal gland) stimulation with hormones or neurotransmitters evokes K+ loss due to opening of K+ channels in the plasma membrane whereas in the insulin-secreting pancreatic beta-cells, stimulation with glucose or glyceraldehyde evokes membrane depolarization due to closure of K+ channels. By measuring directly the small K+ currents flowing through single channels, in electrically isolated patches of plasma membrane of intact cells, it can be shown that stimulants having no direct access to the small membrane area from which recording is made can influence the pattern of channel opening. In the case of hormonal activation of exocrine acinar cells, Ca2+ is the final messenger and the K+-selective channel involved in the response has a high unit conductance, is very voltage sensitive and can be blocked by external tetraethylammonium. In the case of the insulin-secreting cells, the K+ channel which is inhibited by metabolic stimulation is a voltage-insensitive, inward rectifier which can be blocked by quinine. In experiments on permeabilized cells or cell-free excised, inside-out, membrane patches it can be shown that ATP evokes channel closure and ATP produced by glycolysis may therefore function as the internal messenger.

Animals↗
PubMed1985

Voltage-activated Ca2+ currents in insulin-secreting cells.

Membrane voltage and voltage-clamped membrane currents have been investigated with the whole-cell patch clamp method in the insulin-secreting cell line RINm5F. The mean resting membrane potential of RINm5F cells was found to be -52 mV. Overshooting spike potentials could be evoked by depolarising voltage steps in the absence of a secretagogue. Inward membrane currents evoked by depolarising voltage steps were dependent upon extracellular Ca2+ and blocked by Co2+, nifedipine and verapamil. Outward membrane currents which were evoked by depolarising voltage steps to positive membrane potentials were reduced when Ca2+ entry was prevented. It is concluded that the voltage-activated Ca2+ currents underlie the voltage-activated spike potentials recorded from insulin-secreting cells.

Animals↗
PubMed1985

Quinine inhibits Ca2+-independent K+ channels whereas tetraethylammonium inhibits Ca2+-activated K+ channels in insulin-secreting cells.

The effects of quinine and tetraethylammonium (TEA) on single-channel K+ currents recorded from excised membrane patches of the insulin-secreting cell line RINm5F were investigated. When 100 microM quinine was applied to the external membrane surface K+ current flow through inward rectifier channels was abolished, while a separate voltage-activated high-conductance K+ channel was not significantly affected. On the other hand, 2 mM TEA abolished current flow through voltage-activated high-conductance K+ channels without influencing the inward rectifier K+ channel. Quinine is therefore not a specific inhibitor of Ca2+-activated K+ channels, but instead a good blocker of the Ca2+-independent K+ inward rectifier channel whereas TEA specifically inhibits the high-conductance voltage-activated K+ channel which is also Ca2+-activated.

Adenoma, Islet Cell↗
PubMed1985

Acetylcholine stimulates a Ca2+-dependent C1- conductance in mouse lacrimal acinar cells.

Patch-clamp whole-cell current recordings under voltage-clamp conditions were carried out on isolated mouse exorbital lacrimal acinar cells. Acetylcholine evoked outward current at a membrane potential of -20 mV whereas an inward current was observed at -80 mV. The outward current is due to the well-known calcium-activated K+ channels whereas the inward current was C1- dependent. The acetylcholine-evoked C1- current was abolished when the intracellular Ca2+ concentration was clamped at very low levels by a high intracellular EGTA concentration. Acetylcholine therefore activates a Ca2+-dependent C1-conductance in mouse lacrimal acinar cells.

Acetylcholine↗
PubMed1985

ATP-sensitive inward rectifier and voltage- and calcium-activated K+ channels in cultured pancreatic islet cells.

K+ channels in cultured rat pancreatic islet cells have been studied using patch-clamp single-channel recording techniques in cell-attached and excised inside-out and outside-out membrane patches. Three different K+-selective channels have been found. Two inward rectifier K+ channels with slope conductances of about 4 and 17 pS recorded under quasi-physiological cation gradients (Na+ outside, K+ inside) and maximal conductances recorded in symmetrical K+-rich solutions of about 30 and 75 pS, respectively. A voltage- and calcium-activated K+ channel was recorded with a slope conductance of about 90 pS under the same conditions and a maximal conductance recorded in symmetrical K+-rich solutions of about 250 pS. Single-channel current recording in the cell-attached conformation revealed a continuous low level of activity in an apparently small number of both the inward rectifier K+ channels. But when membrane patches were excised from the intact cell a much larger number of inward rectifier K+ channels became transiently activated before showing an irreversible decline. In excised patches opening and closing of both the inward rectifier K+ channels were unaffected by voltage, internal Ca2+ or externally applied tetraethylammonium (TEA) but the probability of opening of both inward rectifier K+ channels was reduced by internally applied 1-5 mM adenosine-5'-triphosphate (ATP). The large K+ channel was not operational in cell-attached membrane patches, but in excised patches it could be activated at negative membrane potentials by 10(-7) to 10(-6) M internal Ca2+ and blocked by 5-10 mM external TEA.

Adenosine Triphosphate↗