Effects of acetylcholine on a slow voltage-activated non-selective cation current mediated by non-nicotinic receptors on isolated Ascaris muscle bags.

Isolated muscle bags from the parasitic nematode Ascaris suum were prepared by collagenase treatment and dissection. Single bags were mounted in a V-shaped plastic pipette for voltage clamp application and intra- and extracellular perfusion. With 'physiological' intra- and extracellular solutions, depolarizing voltage steps from near the normal resting membrane potential, -40 mV, produced in leak-corrected currents, a slowly activating outward current at potentials more positive than -20mV. At the end of the depolarizing pulse there was a slow inward tail current with a reversal potential near -20mV. Hyperpolarizing voltage steps produced an outward current relaxation and an outward tail current with the same reversal potential. The observations can be explained by the presence in the bag of a non-selective cation channel current, Ibcat, that activates spontaneously at the holding potential; depolarization increases opening of the channel and hyperpolarization decreases opening. Bath-applied acetylcholine in concentrations greater than 10(-7) M produced an increase in the amplitude of Ibcat. The effect of acetylcholine was not antagonized or prevented by 100 microM tubocurarine, suggesting the presence of a non-nicotinic acetylcholine receptor. Atropine (100 microM) had no detectable influence on the effect of acetylcholine but the FMRFamide peptide, PF1, in concentrations > 1 microM, reduced the amplitude of Ibcat. Ibcat was maintained when Cs+ was used to replace intra- and extracellular cations, showing that the channels were permeable to Cs+. It is concluded that the bag membrane possesses a slow voltage-activated non-selective cation channel current, Ibcat. The effect of acetylcholine in the presence of nicotinic antagonist indicates the presence of non-nicotinic acetylcholine receptors on the bag membrane. The effect of PF1 indicated the presence of PF1 receptors on the bag membrane.

Electrophysiology of Ascaris muscle and anti-nematodal drug action.

Three groups of anthelmintic drugs act directly and selectively on muscle membrane receptors of parasitic nematodes. These groups of anthelmintics are: (1) The Nicotinic Agonists (levamisole, pyrantel, morantel and oxantel) that act on acetylcholine receptors of nematode somatic muscle; (2) The GABA Agonist, piperazine, that acts on nematode muscle GABA receptors; and (3) The Avermectins that open glutamate gated Cl- channels on nematode pharyngeal muscle. The electrophysiology and pharmacology of muscle and neuromuscular transmission the nematode parasite, Ascaris suum, is outlined and effects of anthelmintics that interfere with transmission described. Resistance to anthelmintics has appeared in some parasitic nematodes but the mechanisms of this resistance remain to be determined.

A Cl channel in Ascaris suum selectivity conducts dicarboxylic anion product of glucose fermentation and suggests a role in removal of waste organic anions.

The permeability of organic anions (produced anaerobic fermentation of glucose) through a non-selective membrane Cl channel was examined. Single channel recording techniques were used to study the permeabilities of the anions: oxalate, succinate, oxaloacetate, malate, lactate and pyruvate in Ascaris muscle cell membranes. All of the anions, except malate, were found to be conducted through the channel. The relative permeability of most anions could be predicted from the component structure of the anions. The failure of the channel to conduct malate prevents an energy drain on the cell. These studies further the hypothesis that a Cl channel functions to transport waste organic anions across the cell membrane. This mechanism does not require specific exchange carriers for the anions. Channels with properties like the nonselective anion channels of Ascaris, are suitable for transport of carboxylic anions through cell membranes, down a concentration or potential gradient.

Transcription of DNA-histone complexes by yeast RNA polymerase B.

Transcription of denatured DNA complexed with histones (total, H1 or H2A/H2B/H3/H4) by yeast RNA polymerase B is investigated. Binding of histones to DNA restricts its template activity by decreasing the formation of active, heparin-resistant, RNA polymerase initiation complexes. The elongation of pre-initiated RNA on denatured DNA, complexed with histones, is possible, although resulting in somewhat shorter RNA chains. It is suggested that RNA polymerase B can elongate on a DNA strand covered with histones.