Extracellular-Regulated Kinases and Phosphatidylinositol 3-Kinase Are Involved in Brain-Derived Neurotrophic Factor-Mediated Survival and neuritogenesis of the Neuroblastoma Cell Line SH-SY5Y

Authors

  • M. Encinas,

    1. Grup de Neurobiologia Molecular, Department de Ciències Mèdiques Bàsiques, Facultat de Medicina, Universitat de Lleida, Lleida, Catalonia, Spain
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  • M. Iglesias,

    1. Grup de Neurobiologia Molecular, Department de Ciències Mèdiques Bàsiques, Facultat de Medicina, Universitat de Lleida, Lleida, Catalonia, Spain
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  • N. Llecha,

    1. Grup de Neurobiologia Molecular, Department de Ciències Mèdiques Bàsiques, Facultat de Medicina, Universitat de Lleida, Lleida, Catalonia, Spain
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  • J. X. Comella

    1. Grup de Neurobiologia Molecular, Department de Ciències Mèdiques Bàsiques, Facultat de Medicina, Universitat de Lleida, Lleida, Catalonia, Spain
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  • The present address of Dr. N. Llecha is Servei d'Anàlisi Clíniques, Hospital Universitari Arnau de Vilanova, Lleida, Catalonia, Spain.

  • Abbreviations used : BDNF, brain-derived neurotrophic factor ; DMEM, Dulbecco's modified Eagle's medium ; ERK, extracellular signal-regulated kinase ; GAP-43, growth-associated protein-43 ; MAPK, mitogen-activated protein kinase ; MEK, MAPK and ERK kinase ; NGF, nerve growth factor ; NT-3, neurotrophin-3 ; NT-4/5, neurotrophin-4/5 ; PAGE, polyacrylamide gel electrophoresis ; PBS, phosphate-buffered saline ; PI 3-K, phosphatidylinositol 3-kinase ; PLCγ, phospholipase Cγ ; RA, retinoic acid ; SDS, sodium dodecyl sulfate ; Trk, tropomyosin receptor kinase.

Address correspondence and reprint requests to Dr. J. X. Comella at Unit of Molecular Neurobiology, Department de Ciències Mèdiques Bàsiques, Universitat de Lleida, Rovira Roure 44, E-25198 Lleida, Spain.

Abstract

Abstract : Retinoic acid (RA) induces the differentiation of many cell lines, including those derived from neuroblastoma. RA treatment of SH-SY5Y cells induces the appearance of functional Trk B and Trk C receptors. Acute stimulation of RA-predifferentiated SH-SY5Y cells with brain-derived neurotrophic factor (BDNF), neurotrophin 3 (NT-3), or neurotrophin 4/5 (NT-4/5), but not nerve growth factor (NGF), induces Trk autophosphorylation, followed by phosphorylation of Akt and the extracellular signal-regulated kinases (ERKs) 1 and 2. In addition, BDNF, NT-3, or NT-4/5, but not NGF, promotes cell survival and neurite outgrowth in serum-free medium. The mitogen-activated protein kinase and ERK kinase (MEK) inhibitor PD98059 blocks BDNF-induced neurite outgrowth and growth-associated protein-43 expression but has no effects on cell survival. On the other hand, the phosphatidylinositol 3-kinase inhibitor LY249002 reverses the survival response elicited by BDNF, leading to a cell death with morphological features of apoptosis.

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