期刊
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH
卷 1499, 期 3, 页码 257-264出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/S0167-4889(00)00127-0
关键词
melatonin; mt(1) receptor; N1E-115 cell; neurite; neuronal development; signal transduction
资金
- NIDDK NIH HHS [DK54070] Funding Source: Medline
- NINDS NIH HHS [NS38873, NS37672] Funding Source: Medline
Melatonin, a pineal hormone that induces sleep, has become a popular over-the-counter drug. The cellular effects of melatonin, however, are only beginning to be studied. We have recently shown that stimulation of the mt(1) melatonin receptor induces rapid and dramatic cytoskeletal rearrangements in transformed non-neuronal cells (Witt-Enderby et al., Cell. Motil. Cytoskel. 46 (2000) 28). These cytoskeletal changes result in the formation of structures that closely resemble neurites. In this work, we show that the N1E-115 mouse neuroblastoma cell line rapidly responds to melatonin stimulation and forms neurites within 24 h. We also demonstrate that these cells readily bind 2-[I-125]iodomelatonin at levels consistent with what is noted for native tissues (B-max = 3.43 +/- 1.56 fmol/mg protein; K-d = 240 pM). Western analysis shows that these cells possess and express melatonin receptors of the mt(1) subtype. Treatment with pertussis toxin eliminates neurite formation whereas treatment with the MT2 subtype-specific activator, BMNEP, does not induce neurite formation. We have previously shown that increases in MEK 1/2 and ERK 1/2 phosphorylation are correlated with the shape changes in transformed CHO cells. Western analysis of the MEK/ERK signaling pathway in N1E-115 cells shows that this pathway is most likely maximally and constitutively stimulated. This may account for the spontaneous production of neurites noted for this cell line after long culture periods. The results of this work show that melatonin receptor stimulation in a neuronal cell type results in the formation of neurites and that the receptors responsible for melatonin-induced neurite formation in N1E-115 cells are most likely of the mt(1) subtype. (C) 2001 Elsevier Science B.V. All rights reserved.
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