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Endocrinology Vol. 141, No. 1 412-419
Copyright © 2000 by The Endocrine Society

ARTICLES

Genetic Targeting of Green Fluorescent Protein to Gonadotropin-Releasing Hormone Neurons: Characterization of Whole-Cell Electrophysiological Properties and Morphology1

Kelly J. Suter, Walter J. Song, Traci L. Sampson, Jean-Pierre Wuarin, Justin T. Saunders, F. EDWARD Dudek and Suzanne M. Moenter

Department of Anatomy and Neurobiology (K.J.S., T.L.S., J.-P.W., F.E.D.), Animal Reproduction and Biotechnology Laboratory (K.J.S.), Colorado State University, Fort Collins, Colorado 80523; and Departments of Internal Medicine and Cell Biology (W.J.S., J.T.S., S.M.M.), National Science Foundation Center for Biological Timing, University of Virginia, Charlottesville, Virginia 22908

Address all correspondence and requests for reprints to: Suzanne M. Moenter, Department of Medicine, Box 578 HSC, University of Virginia, Charlottesville, Virginia 22908. E-mail: smm4n{at}virginia.edu

GnRH neurons form the final common pathway for central control of reproduction, with regulation achieved by changing the pattern of GnRH pulses. To help elucidate the neurobiological mechanisms underlying pulsatile GnRH release, we generated transgenic mice in which the green fluorescent protein (GFP) reporter was genetically targeted to GnRH neurons. The expression of GFP allowed identification of 84–94% of immunofluorescently-detected GnRH neurons. Conversely, over 99.5% of GFP-expressing neurons contained immunologically detectable GnRH peptide. In hypothalamic slices, GnRH neurons could be visualized with fluorescence, allowing for identification of individual GnRH neurons for patch-clamp recording and subsequent morphological analysis. Whole-cell current-clamp recordings revealed that all GnRH neurons studied (n = 23) fire spontaneous action potentials. Both spontaneous firing (n = 9) and action potentials induced by injection of depolarizing current (n = 17) were eliminated by tetrodotoxin, indicating that voltage-dependent sodium channels are involved in generating action potentials in these cells. Direct intracellular morphological assessment of GnRH dendritic morphology revealed GnRH neurons have slightly more extensive dendrites than previously reported. GnRH-GFP transgenic mice represent a new model for the study of GnRH neuron structure and function, and their use should greatly increase our understanding of this important neuroendocrine system.




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