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Evidence that Brain-Derived Neurotrophic Factor Acts as an Autocrine Factor on Pituitary Melanotrope Cells of Xenopus laevis

Departments of Cellular Animal Physiology (B.M.R.K., P.M.J.M.C., D.T.W.M.O., E.W.R., B.G.J.) and Molecular Animal Physiology (M.W.C., G.J.M.M.), Nijmegen Institute for Neurosciences and Institute of Cellular Signalling, University of Nijmegen, Nijmegen 6525 ED, The Netherlands

Address all correspondence and requests for reprints to: Bruce G. Jenks, Department of Cellular Animal Physiology, Nijmegen Institute for Neurosciences, University of Nijmegen, Toernooiveld 1, Nijmegen 6525 ED, The Netherlands. E-mail: . jenks{at}sci.kun.nl

We have investigated the physiological regulation and functional significance of brain-derived neurotrophic factor (BDNF) in the endocrine melanotrope cells of the pituitary pars intermedia of the amphibian Xenopus laevis, which can adapt its skin color to the light intensity of its environment. In black-adapted animals, melanotrope cells produce and release -melanophore-stimulating hormone (-MSH). In white-adapted animals, the activity of melanotrope cells is inhibited by neuronal input. Using Western blotting and immunocytochemistry at the light and electron microscopical level, we have detected both the BDNF precursor and the mature BDNF protein in Xenopus melanotrope cells. In situ hybridization and RT-PCR revealed the presence of BDNF mRNA in the pituitary pars intermedia, indicating that BDNF is synthesized in the melanotropes. Real-time quantitative RT-PCR showed that levels of BDNF mRNA in melanotrope cells are about 25 times higher in black- than in white-adapted animals. Although there is no difference in the amount of stored mature BDNF, the amount of BDNF precursor protein is 3.5 times higher in melanotropes of black-adapted animals than in those of white-adapted animals. These data indicate that BDNF mRNA expression and BDNF biosynthesis are up-regulated in active melanotrope cells. Because immunoelectron microscopy showed that BDNF is located in melanotrope secretory granules, BDNF is probably coreleased with -MSH via the regulated secretory pathway. Superfusion and ³H-amino acid incorporation studies demonstrated that BDNF stimulates the release of -MSH and the biosynthesis of its precursor protein, POMC. Our results provide evidence that BDNF regulates the activity of Xenopus melanotrope cells in an autocrine fashion.

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