The secretion of luteinizing hormone (LH) is cued by the hypothalamic neuropeptide, gonadotropin-releasing hormone (GnRH). Following delivery to the anterior pituitary gland via the hypothalamic-pituitary portal vasculature, GnRH binds to specific high affinity receptors on the surface of gonadotrope cells and stimulates synthesis and secretion of the gonadotropins, follicle stimulating hormone (FSH) and LH. In the current study, GnRH caused acute and dramatic changes in cellular morphology in the gonadotrope derived T3-1 cell line, which appeared to be mediated by engagement of the actin cytoskeleton; disruption of actin with jasplakinolide abrogated cell movement and GnRH induced activation of ERK. In live murine pituitary slices infected with an adenovirus containing Rous Sarcoma Virus-Green Fluorescent Protein (RSV-GFP), selected cells responded to GnRH by altering their cellular movements characterized by both formation and extension of cell processes and, surprisingly, spatial repositioning. Consistent with the latter observation, GnRH stimulation increased the migration of dissociated pituitary cells in transwell chambers. Our data using live pituitary slices is a striking example of neuropeptide-evoked movements of cells outside of the CNS and in a mature peripheral endocrine organ. These findings call for a fundamental change in the current dogma of simple passive diffusion of LH from gonadotropes to capillaries in the pituitary gland.