William R. Harvey
(Ph.D. Harvard, 1957)

Professor of Physiology and Functional Genomics,
Editor, Journal of Experimental Biology

wharvey@whitney.ufl.edu

Membrane Physiology

Our goal is to understand how solutes move into and out of epithelial cells. In particular, we study how animals take up nutrient amino acids and ions, such as Na+, K+, and Cl-, from the digestive tract. In the pre-genome era we used the highly alkaline caterpillar midgut as a model. We have isolated and characterized a membrane transport protein that is now known as a proton-translocating, vacuolar-type ATPase (H+ V-ATPase). The V-ATPase imposes a voltage across midgut epithelial cell membranes and the voltage in turn drives ion exchange and nutrient uptake via other proteins called transporters. We have cloned all subunits of the caterpillar H+ V-ATPase and learned much about its structure and function. We have also cloned two amino acid transporters from caterpillar midgut.

In the post-genomic era we have developed a new model – the larval midgut of two disease vector mosquitoes, the African malaria mosquito, Anopheles gambiae and the yellow fever mosquito, Aedes aegypti. Since the Spring of 2002, when the Anopheles mosquito genome became available, we have identified sixteen amino acid transporters and several salt transporters. The An. gambiae midgut is proving a excellent preparation for understanding solute transport in mosquito larvae and a useful model for solute transport across other epithelia, such as those in human kidney, eye, and bone.

Current Projects

Transport physiology of disease vector mosquitoes. Bernard Okech uses techniques in molecular biology and immuno-cytochemistry to identify proteins and other factors within the alimentary canal and blood that influence the survival of insect and the pathogens they carry. In addition, He also employs high resolution microscopy and histological techniques to study the cellular factors involved in nutrient absorption and other regulatory processes in mosquito gut that enable survival of the vector and pathogen.

Anion regulation in larvae of disease vector mosquitoes. I collaborate with Paul Linser and his associates in determining how the V-ATPase-generated voltage drives bicarbonate ions into the lumen while driving chloride ions to the blood. We have shown that this exchange could help account for the alkalinity of larval midgut and are searching for other ion transporters that could account for a pH that has been shown to reach values as high as 11. Dr. Linser’s group has cloned several carbonic anhydrases that are involved in generating the bicarbonate plus a chloride/bicarbonate exchanger.

Personnel

William R. Harvey, Professor, Editor,Journal of Experimental Biology
Bernard Okech, Research Assistant Professor

Selected Publications

Meleshkevitch, E.A., Assis-Nascimentol, P., Popova, L.B., Miller, M.M., Kohn, A.B., Phung, L., Mandal, A., Harvey, W.R. and Boudko, D.Y. (2006) Molecular characterization of the first aromatic nutrient transporter from the sodium neurotransmitter symporter family, J. Exp. Biol. In press.

Boudko D.Y., Kohn A.B., Meleshkevitch E.A., Dasher M.K., Stevens B.R. and Harvey W.R. (2005)
Ancestry and progeny of nutrient transporters, Proc. Natl. Acad. Sci. U.S.A. 102, 1360-1365.

Boudko, D.Y., Stevens, B.R., Donly, B. C. and Harvey, W.R. (2005) Amino acid and neurotransmitter transporters, In Comprehensive Insect Science, Vol. 5, Pharmacology, Editors: Lawrence I. Gilbert, Kostas Iatrou, and Sarjeet Gill. Elsevier Pergamon, Amsterdam pp 2555-309.

Liu, Z., Stevens, B.R., Feldman, D.H., Hediger, M.A. and Harvey, W.R. (2003) K+ amino acid transporter KAAT1 mutant Y147F has increased transport activity and altered substrate selectivity. J. Exp. Biol. 206, 245-254.

Liu, Z., Stevens, B.R., Feldman, D.H., Hediger, M.A. and Harvey, W.R. (2002). K+ amino acid transporter KAAT1 Mutant Y147F has increased transport activity and altered substrate selectivity. J. Exp. Biol. 206, 245-254.

Boudko, D.Y., Cooper, B.Y., Harvey, W.R. and Moroz, L.L. (2002). High-resolution microanalysis of nitrite and nitrate in neuronal tissues by capillary electrophoresis with conductivity detection. J. Chromatogr. B. 774: 97-104.

Stevens, B. R., Feldman, D.H., Liu, Z., and Harvey, W.R. (2002) Conserved tyrosine-147 plays a critical role in the ligand-gated current of the epithelial cation/amino acid transporter/channel CAATCH1. J. Exp. Biol. 205:2545-2553.

Boudko, D.Y., Moroz, L.L., Harvey, W.R. and Linser, P.J. (2001), Alkalinization by chloride/ bicarbonate pathway in larval mosquito midgut. Proc. Natl. Acad. Sci. U.S.A. 98 15354-15359.

Grüber, G., Wieczorek, H., Harvey, W.R. and Müller, V. (2001), Structure-Function relationships of A1-, F1-, and V1-ATPases. J. Exp. Biol. 204, 2597-2605.

Boudko, D.Y., Moroz, L.L., Linser, P.J. Trimarchi, J.R., Smith, P.J.S. and Harvey, W.R.(2001), In situ analysis of pH gradients in mosquito larvae using non-invasive, self-referencing, pH-sensitive microelectrodes. J. Exp. Biol. 204, 691-699.

Stevens, B.R., Tellier, M., Harvey, W., Feldman, D.H., and Bosworth, J. 2000. Interleukin-2 and concanavalin A upregulate a CAT2 isoform encoding a high affinity L-arginine transporter in feline lymphocytes. Can. J. Vet. Res. 64:187-191.

Merzendorfer, H., Reineke, S., Zhao, X-F, Jacobmeier, B. Harvey, W.R. and Wieczorek, H. (2000). The multigene family of the tobacco hornworm V-ATPase: novel subunits a, C, D, H, and putative isoforms. Biophys. Biochim. Acta 1467, 369-379.

Grüber, G., Svergun, D.I., Godovac-Zimmermann, J. Harvey, W.R., Wieczorek, H. and Koch, M.H.J. (2000), Evidence for major structural changes in the Manduca sexta midgut V1 ATPase due to redox modulation A small angle X-ray scattering study. J. Biol. Chem. 275, 30082-30087.

Grüber, G., Radermacher, M., Ruiz, T., Godovac-Zimmermann, J., Canas, B., Kleine-Kohlbrecher, D., Harvey, W.R. and Wieczorek, H. (2000). Three-dimensional structure and subunit topology of the V1 ATPase from Manduca sexta. Biochemistry 39, 8609-8616.

Feldman, D. H., Harvey, W.R. and Stevens, B.R. (2000). A novel electrogenic amino acid transporter is activated by K+ or Na+, is alkaline pH-dependent, and is Cl-independent. J. Biol. Chem. 275, 24518-24526.

Wieczorek, H., Grüber, G., Harvey, W.R., Huss, M., Merzendorfer, H. and Zeiske, W. (2000), Structure and regulation of insect plasma membrane H+ V-ATPase. J. Exp. Biol. 203, 127-135.

Zhuang, Z., Linser, P.J. and Harvey, W.R. (1999). Antibody to H+ V-ATPase subunit E colocalizes with portasomes in alkaline larval midgut of a freshwater mosquito (Aedes aegypti L.). J.Exp. Biol. 202, 2449-2460.

Radermacher, M., Ruiz, T., Harvey, W.R., Wieczorek, H. and Grüber, G. (1999), Molecular architecture of Manduca sexta midgut V1ATPase visualized by electron microscopy. FEBS Lett. 453, 383-386.

Merzendorfer, H., Huss, M., Schmid, R., et al. (1999). A novel insect V-ATPase subunit M9.7 is glycosylated extensively. J. Biol. Chem. 274, 17372-17378.

Wieczorek, H., Grüber, G., Harvey, W.R., Huss, M., and Merzendorfer, H. (1999), The plasma membrane H+ V-ATPase from tobacco hornworm midgut. J. Bioenerg. Biomembr. 31, 67-74.

Nelson, N. and Harvey, W.R. (1999), Vacuolar and plasma membrane proton adenosine triphosphatases. Physiol. Rev. 79, 361-385.

Wieczorek, H., Brown, D., Grinstein, S., Ehrenfeld, J. and Harvey, W.R. (1999), Animal plasma membrane energization by proto-motive V-ATPases. BioEssays, 21, 637-648.

Harvey, W.R., Maddrell, S.H.P., Telfer, W.H. and Wieczorek, H. (1998) H+ V-ATPases energize animal plasma membranes for secretion and absorption of ions and fluids. Amer. Zool. 38, 426-441.

Castagna, M., Shayakul, C., Trotti, D., Sacchi, V.F., Harvey, W.R. and Hediger, M.A. (1998), Cloning and characterization of a potassium-coupled amino acid transporter. Proc. Natl. Acad. Sci., USA 95, 5395-5400.