K+ transport in Malpighian tubules ofTenebrio molitorL.: a study of electrochemical gradients and basal K+uptake mechanisms

Abstract

SUMMARYMalpighian tubules of the mealworm Tenebrio molitor were isolated for intracellular measurement of basolateral (Vbl) and,indirectly, apical (Vap) membrane potentials. In control Ringer (50 mmol l-1 K+, 140 mmol l-1Na+), Vbl was 24mV, cell negative, and Vap was 48 mV, cell negative with reference to the lumen. Ion substitution experiments involving K+ and Na+indicated that both Vbl and Vap were sensitive to the bathing K+ concentration, with the change in Vap being 60-77% that of Vbl. A 10-fold drop in bath [K+] irreversibly decreased fluid secretion rates from 6.38±0.95 nl min-1 (mean ± S.E.M.) to 1.48±0.52 nl min-1 (N=8). In the presence of 6 mmol l-1 Ba2+, a blocker of basal K+ channels,fluid secretion rates reversibly decreased and the hyperpolarization of both Vbl and Vap seen in 50 mmol l-1 and 140 mmol l-1 K+ indicated a favourable electrochemical gradient for basal K+ entry. In 5 mmol l-1 K+, Ba2+ induced two different responses: Vbl either hyperpolarized by approximately 10 mV or depolarised by approximately 14mV, according to the electrochemical gradient for K+, which was either inward or outward in low bath[K+]. Rubidium, a `permeant' potassium substitute, caused a hyperpolarization of Vbl, indicating the specificity of K+ channels found in Tenebrio tubule cells. Other possible K+ uptake mechanisms located in the basolateral membrane were investigated. Blocking of the putative electroneutral Na+/K+/2Cl- cotransporter by 10 μmol l-1 bumetanide reversibly decreased fluid secretion rates, with no detectable change in membrane potentials. Ouabain (1 mmol l-1), an Na+/K+-ATPase inhibitor, irreversibly decreased fluid secretion rates but had no effect on electrical potential differences either in the absence or presence of Ba2+. The results implicate K+ channels, the Na+/K+/2Cl-contransporter and the Na+/K+-ATPase in basal K+ and fluid transport of Tenebrio tubule cells.