Genetic targeting of green fluorescent protein to gonadotropin-releasing hormone neurons: characterization of whole-cell electrophysiological properties and morphology.
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.
Suter
K J
KJ
Department of Anatomy and Neurobiology, Colorado State University, Fort Collins 80523, USA.
Song
W J
WJ
Sampson
T L
TL
Wuarin
J P
JP
Saunders
J T
JT
Dudek
F E
FE
Moenter
S M
SM
D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D020667:Diagonal Band of Broca / Q000166:cytology / Q000378:metabolism; D004594:Electrophysiology; D019085:Fluorescent Antibody Technique, Direct; D005786:Gene Expression Regulation / Q000502:physiology; D018390:Gene Targeting; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000235:genetics*; D051379:Mice; D009474:Neurons / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D018408:Patch-Clamp Techniques; D013779:Tetrodotoxin / Q000494:pharmacology; D019076:Transgenes / Q000235:genetics
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D007987:Gonadotropin-Releasing Hormone; D013779:Tetrodotoxin