Evidence That the ZNT-3 Protein Controls the Total Amount of Elemental Zinc in Synaptic Vesicles

D.H. Linkous ¹, J.M. Flinn ¹, J.Y. Koh ¹, A. Lanzirotti ¹, P. Bertsch ¹, B.F. Jones ¹, L.J. Giblin ¹ and C.J. Frederickson ^1*

¹ Department of Psychology, George Mason University, Fairfax, Virginia (DHL,JMF); Department of Neurology, University of Ulsan College of Medicine, Seoul, Korea (JYK); Consortium for Advanced Radiation Sources, University of Chicago, Chicago, Illinois (AL); National Synchrotron Light Source, Brookhaven National Laboratory, Upton, New York (AL); Savannah River Ecology Laboratory, University of Georgia, Aiken, South Carolina (PB); United States Geological Survey, Reston, Virginia (BFJ); NeuroBioTex, Inc., Galveston, Texas (LJG,CJF)

^* To whom correspondence should be addressed. E-mail: c.j.frederickson{at}neurobiotex.com.

Submitted on June 15, 2006
Accepted on 18 July 2007

	Abstract

The ZNT3 protein decorates the presynaptic vesicles of central neurons harboring vesicular zinc, and deletion of this protein removes staining for zinc. However, it has been unclear whether only histochemically reactive zinc is lacking or if indeed total elemental zinc is missing from neurons lacking the Slc30a3 gene, which encodes the ZNT3 protein. The limitations of conventional histochemical procedures have contributed to this enigma. However, a novel technique, microprobe synchrotron X-ray fluorescence (µSXRF), reveals that the normal 2- to 3-fold elevation of zinc concentration normally present in the hippocampal mossy fibers is absent in Slc30a3 (ZNT3) knockout mice. Thus, the ZNT3 protein evidently controls not just the "stainability" but also the actual mass of zinc in mossy fiber synaptic vesicles. This work thus confirms the metal transporting role of the ZNT3 protein in the brain.

Key Words: mossy fibers, ZNT3, glutamate, zinc release, hilus, x-ray, fluoresence, knock-out

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