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Am J Physiol Renal Physiol 279: F747-F754, 2000;
0363-6127/00 $5.00
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Vol. 279, Issue 4, F747-F754, October 2000

Voltage clamping single cells in intact Malpighian tubules of mosquitoes

R. Masia1, D. Aneshansley2, W. Nagel3, R. J. Nachman4, and K. W. Beyenbach1

Departments of 1 Biomedical Sciences and 2 Agricultural and Biological Engineering, Cornell University, Ithaca, New York 14853; 3 Department of Physiology, University of Munich, D-80336 Munich, Germany; and 4 VERU, Southern Plains Agricultural Research Center, United States Department of Agriculture, College Station, Texas 77845

Principal cells of the Malpighian tubule of the yellow fever mosquito were studied with the methods of two-electrode voltage clamp (TEVC). Intracellular voltage (Vpc) was -86.7 mV, and input resistance (Rpc) was 388.5 kOmega (n = 49 cells). In six cells, Ba2+ (15 mM) had negligible effects on Vpc, but it increased Rpc from 325.3 to 684.5 kOmega (P < 0.001). In the presence of Ba2+, leucokinin-VIII (1 µM) increased Vpc to -101.8 mV (P < 0.001) and reduced Rpc to 340.2 kOmega (P < 0.002). Circuit analysis yields the following: basolateral membrane resistance, 652.0 kOmega ; apical membrane resistance, 340.2 kOmega ; shunt resistance (Rsh), 344.3 kOmega ; transcellular resistance, 992.2 kOmega . The fractional resistance of the apical membrane (0.35) and the ratio of transcellular resistance and Rsh (3.53) agree closely with values obtained by cable analysis in isolated perfused tubules and confirm the usefulness of TEVC methods in single principal cells of the intact Malpighian tubule. Dinitrophenol (0.1 mM) reversibly depolarized Vpc from -94.3 to -10.7 mV (P < 0.001) and reversibly increased Rpc from 412 to 2,879 kOmega (P < 0.001), effects that were duplicated by cyanide (0.3 mM). Significant effects of metabolic inhibition on voltage and resistance suggest a role of ATP in electrogenesis and the maintenance of conductive transport pathways.

yellow fever mosquito; shunt resistance; potassium channel; barium; leucokinin


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