Abstract
Cellular distribution, receptor‐receptor interaction, and signaling pathways of the murine GnRH‐R tagged with Renilla luciferase (Rluc) and/or green fluorescent protein (GFP) were monitored in immortalized (GT1‐7) GnRH neurons. GFP‐tagged GnRH‐R was confined to a thin rim of cytosol at the plasma membrane, neuronal processes, and apparent synaptic junctions. GnRH stimulation caused redistribution of GnRH‐R‐GFP, with movement in close proximity to their bipolar extensions and increased intensity in synaptic connections. BRET2 assay revealed dose‐dependent, GnRH‐induced interaction between GFP‐tagged GnRH‐R and Rluc‐tagged Gαq, Gαs and Gαi. Firing of spontaneous action potentials (APs), calcium signaling and pulsatile GnRH secretion were comparable to those of intact GT1‐7 neurons. These findings indicate that specific subcellular localization, agonist‐induced redistribution, homologous oligomerization, and coupling to multiple G proteins of the GnRH receptor, modulate electrical activity, second messengers, and neurosecretion in hypothalamic GnRH neurons.
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