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Journal of Experimental Biology, Vol 197, Issue 1 421-428, Copyright © 1994 by Company of Biologists
JOURNAL ARTICLES |
JAT. Dow, SHP. Maddrell, A. Gorrtz, NJV. Skaer, S. Brogan and K. Kaiser
The insect renal (Malpighian) tubule has long been a model system for the study of fluid secretion and its neurohormonal control (Maddrell, 1981; Maddrell and O'Donnell, 1992). Classical physiology suggests a model for tubular secretion of iso-osmotic fluid in most insects, in which ions are thought to enter basally either through a series of ion channels (Na+, K+ and Cl-) or through a bumetanide-sensitive Na+/K+/2Cl- cotransport. Apical fluxes are energised by a plasma-membrane H+-pumping V- ATPase, driving secretion of Na+ or K+ through one or more exchangers, at least one of which is amiloride-sensitive and appears to be closely similar to the Na+/H+ exchanger of vertebrates (Maddrell and O'Donnell, 1992). Cl- follows passively, perhaps through apical Cl- channels. Water follows the major ions, and haemolymph solutes diffuse across the tubule wall passively via a paracellular route. There are also transcellular active transport processes for certain metabolites or toxins, such as acylamides (Maddrell et al. 1974) and plant alkaloids (Maddrell, 1976; O'Donnell et al. 1983). To extend studies of ion transport beyond that revealed by the techniques of classical physiology, it is necessary to adopt a molecular genetic approach, in which the relevant genes are identified, characterised and mutated. However, the generalised molecular genetic dissection of the major genes responsible for a stimulus-secretion pathway in any vertebrate epithelium remains a daunting task, as the genetic tools available are relatively unsophisticated. Encouraged by previous physiological work on the larger, but less well genetically mapped, Drosophila hydei (Bertram et al. 1991; Wessing and Eichelberg, 1978; Wessing et al. 1987), we have investigated the feasibility of adapting the established tubule secretion assay to the study of tubules of larval and adult Drosophila melanogaster. The results (i) demonstrate that these Malpighian tubules, to our knowledge the smallest yet studied, are amenable to physiological study; (ii) describe a novel modification of the tubule assay system that allows virtually the entire tubule length to be employed in secretion studies; and (iii) show that the epithelial transport mechanisms identified in D. melanogaster tubules are comparable with those in tubules of other insect species. We propose, therefore, a complete physiological and molecular dissection of this tissue, which may not be possible in any other transporting epithelium.
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