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Biomedical subjects

A W Cuthbert

Publications and source records attributed to A W Cuthbert.

At least 127 records · Page 7Linked to original sources
PubMed1978

Effects of some pyrazinecarboxamides on sodium transport in frog skin.

1 The inhibitory effect of amiloride (N-amidino-3,5-diamino-6-chloropyrazinecarboxamide) on sodium transport in isolated skin of frog has been compared with 17 of its analogues. The dissociation constant of amiloride for passive sodium channels was 181.9 +/- 8.9 nM, and the maximal percentage inhibition of sodium transport was 101.3 +/- 0.4% (means of 123 measurements) when measured at a sodium concentration of 111 mM. 2 The N-benzylamidino and N-o-chlorobenzylamidino compounds had affinities approximately 20 times larger than those for amiloride, and produced maximal inhibition of transport. 3 Substitution of chlorine in the 6-position by other halogens showed that the bromo-compound was equally active to amiloride, whereas the iodo derivative had an affinity equal to 15% of that for amiloride. 4 Substitution in the 5-amino group in 10 compounds reduced the affinities to less than 1% of that of amiloride, without affecting their ability to produce complete inhibition of transport. 5 N-Amidino-3,5-diaminopyrazinecarboxamide was unique in that it produced an unusual concentration-response relationship.

Amiloride↗
PubMed1976

Similarities between sodium channels in excitable membranes and in epithelia.

The inhibitory effects of the pyrazine derivative, amiloride, on sodium transport in an amphibian epithelium has been studied as a function of pH. It is concluded that the charged (guanidinium) group interacts with a negatively charged acid grouping in the membrane. Similarities between sodium channels in excitable membranes and epithelia are highlighted.

Amiloride↗
PubMed1976

Characteristics of the entry process for sodium in transporting epithelia as revealed with amiloride.

1. The permeation of sodium ions trhough the mucosal surface of frog skin epithelium at different transepithelial potentials has been investigated using the blocking drug amiloride. 2. An increase in serosal negativity in voltage-clamped skins was associated with an increase in the absolute amount of inhibition caused by a fixed concentration of amiloride. Hyperpolarizing or depolarizing skins with respect to the short-circuited condition did not affect the apparent affinity of amiloride for the entry sites. 3. When skins were voltage clamped at -50 mV (serosa negative) the specific binding of amiloride to sodium entry sites was increased by 77% compared to the short-circuited condition. Skins clamped at +50 mV had only 72% of the specific binding found in short-circuited skins. Experiments with a second blocking drug, triamterene, indicated that the extra binding sites appearing at -50mV were similar to those found under short-circuit conditions. The appearance and disappearance of binding sites may reflect changes in cell volume. 4. The findings suggest that the increased sodium current which flows when skins are clamped at -50 mV results from an increase in the number of entry sites, and perhaps also to a voltage sensitive increase in flux through each entry site.

Amiloride↗
PubMed1976

Estimation of the lifespan of amiloride binding sites in the membranes of toad bladder epithelial cells.

1. Sodium entry sites in the membranes of isolated epithelial cells prepared from bladders of toads (Bufo marinus) have been labelled with amiloride. The number of binding sites remained constant in suspensions for up to 100 hr. 2. In the presence of a protein synthesis inhibitor (cycloheximide, 0-5 mug/ml.) there was a decline in the density of binding sites was approximately exponential. Regression analysis gave a half-life of approximately 60 hr. 3. Aldosterone (5 X 10(-8) M) caused a significant (P less than 0-001) increase (50%) in the density of amiloride binding sites. Cells which had been treated with aldosterone had populations of binding sites which declined, in the presence of cycloheximide, at rates indistinguishable from those of untreated cells.

Aldosterone↗
PubMed1976

Induction of transporting sites in a sodium transporting epithelium.

1. Frogs (Rana temporaria) were bathed for 1 week in solutions containing 1-1 mM sodium chloride and either one or both of amiloride (10(-4)M) and spironolactone (10(-5r both of amiloride (10(-4) M) and spironolactone (10(-5) M). This procedure was designed to deplete the sodium transporting compartment of the skin epithelium of sodium, while at the same time antagonizing the effects of endogenous aldosterone. 2. After 1 week the skins were used in vitro to measure the level of sodium transport (short-circuit current) and the density of sodium entry sites in the mucosal surface of the epithelium ([14C]amiloride binding). 3. Sodium deprivation for 1 week caused approximately a doubling of both sodium transport and the density of sodium entry sites in the mucosal surface of the epithelium compared to control skins. 4. When the results for sodium deprived and control skins were pooled there was a highly significant correlation between the density of sodium entry sites and sodium transport. 5. Mechanisms by which sodium deprivation leads to an increase in the density of sodium entry sites are discussed.

Amiloride↗
PubMed1975

Muscarinic receptors in rat intestinal muscle: comparison with the guinea pig.

The properties of muscarinic receptors in rat intestinal muscle have been investigated by examining the binding of 3H-propylbenzilylcholine mustard, 3H-PrBCM, a specific and virtually irreversible muscarinic antagonist. It is shown that the properties of these muscarinic receptore correspond in general closely to those observed in the guinea pig, in particular the binding curve inferred for carbachol shows the same apparent negative cooperativity, but there is a notable difference in the rate of binding 3H-PrBCM.

Animals↗
PubMed1975

Intracellular potentials in cells of the seminiferous tubules of rats.

1. Membrane potentials have been recorded from cells of seminiferous tubules of rats in vitro using micro-electrodes. The value in 808 impalements was -28-2 +/- 0-3 mV (mean +/- S.E.) at 33 degrees C. 2. Increasing the potassium concentration depolarized the cells, a tenfold increase in concentration causing a depolarization of 16 mV. Removal of sodium from the bathing solution caused a hyperpolarization of 3 mV at a potassium concentration of 5-9 m-equiv/l. Removal of chloride and replacement with impermeant anions had no effect on potential. Removal of calcium from the bathing solution caused a minor but significant depolarization. 3. Ouabain (10-3 M), dinitrophenol (2-5 times 10-4 M) or removal of glucose from the bathing fluid all caused depolarization. The membrane potentials of the cells were sensitive to temperature over the range 10-33 degrees C, the apparent activation energy for the reactions maintaining the potential being approximately 6 kcal/mole. 4. Membrane potentials in seminiferous tubules were independent of age of the animal, were insensitive to previous hypophysectomy and were insensitive to a number of hormones (FSH, LH, HCG, oxytocin). In high concentration prostaglandin E1 caused depolarization. 5. Acetazoleamide (4 times 10-5 M) caused a rapid, but reversible, depolarization of the tubular cells. This was also true in conditions when the HCO'3/CO2 buffer system was replaced with Tris-buffer. Another carbonic anhydrase inhibitor (p-sulphonamido-benzoic acid) had similar effects on cell potentials as acetazoleamide. These results are discussed in relation to the nature of the ionic secretion produced in the tubules. 6. Occasional cells showed phasic variations in membrane potential. A possible connexion between these variations and the contractile activity of the tubules is discussed.

Acetazolamide↗
PubMed1974

Calcium release in relation to permeability changes in toad bladder epithelium following antidiuretic hormone.

1. Methods for measuring the release of (45)Ca from isolated urinary bladders of toads (Bufo marinus) pre-loaded with this isotope have been devised. One method allowed separate collection from the mucosal and serosal surfaces of the bladders.2. Reducing the ambient calcium concentration reduced the rate of (45)Ca efflux suggesting that efflux of radiolabel represents calcium exchange.3. Antidiuretic hormone, theophylline and prostaglandin E(1) all increased calcium efflux, while lanthanum and amphotericin were without effect. Cyclic AMP caused only an inhibition of calcium release.4. The increase in (45)Ca efflux due to antidiuretic hormone came exclusively from the mucosal side. Experiments with EGTA suggest that the calcium entering the mucosal solution arises mainly from superficial sites in the mucosal membrane.5. The release of (45)Ca by hormone was not influenced by removal of sodium from the bathing solution. Low pH and amiloride reduced or abolished calcium release to hormone.6. The time course of calcium release from the mucosal surface due to hormone was rapid (commencing between 0.5 and 1.5 min after hormone application). Thus calcium release precedes the increase in sodium transport and hydro-osmotic flow following hormone, and appears to be at least as rapid as cyclic AMP generation in the tissue.7. The relationship between calcium release or exchange and the permeability changes in the bladder to water and to sodium, following hormone, are discussed.

Amiloride↗