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On the distribution of a fluorescent ivermectin probe (4″ 5,7 dimethyl-bodipy proprionylivermectin) in Ascaris membranes

Published online by Cambridge University Press:  06 April 2009

R. J. Martin  and
J. R. Kusel
R. J. Martin
Affiliation:
Department of Pre-clinical Veterinary Sciences, R.(D.) S.V.S. Summerhall, University of Edinburgh, Edinburgh EH9 IQH
J. R. Kusel
Affiliation:
Department of Biochemistry, University of Glasgow, Glasgow
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Extract

The distribution of the fluorescent ivermectin derivative (4″ 5,7 dimethyl bodipy proprionylivermectin, referred to here as bodipy ivermectin) was studied in muscle vesicle membranes from the parasite Ascaris suum. Incubation in bodipy ivermectin, at concentrations greater than 1 ng/ml, produced a dose-dependent fluorescence in the vesicles. The level of fluorescence was not depressed (but increased) by prior incubation with 1 μM unlabelled ivermectin. This observation suggested that most of the bodipy ivermectin was free in the lipid phase of the membrane and not associated with a receptor. The trypan blue quenching technique showed that the bodipy ivermectin did not cross the membrane bilayer but remained in the outer monolayer. Lateral mobility of the probe in the membrane was studied using the fluorescence recovery after photobleaching (FRAP) technique. The lateral diffusion coefficient (DL) was 14.0 x 10−10 cm2/s which was nearly three times faster than the lipid probe octadecanoyl-aminofluorescein in membranes from the same preparations; only 46% of the probe was mobile, showing that within a 1.2 μm radius the distribution of the bodipy ivermectin in the membrane was not homogeneous. These observations are used to suggest that: the site of action of livermectin is located in the outer monolayer of the membrane (possibly on the Cl− channel); and that the speed of onset of action in isolated membrane preparations may be limited by diffusion in the membrane.

Keywords

Ascaris suum4″ 5,7 dimethyl bodipy proprionylivermectinbodipy ivermectinmembrane distribution
Type
Research Article
Information
Parasitology , Volume 104 , Issue 3 , June 1992 , pp. 549 - 555
Copyright
Copyright © Cambridge University Press 1992

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