Differentiation of mouse Neuro 2A cells into dopamine neurons
作者： Roger G. Tremblay , Marianna Sikorska , Jagdeep K. Sandhu , Patricia Lanthier , Maria Ribecco-Lutkiewicz
DOI: 10.1016/J.JNEUMETH.2009.11.004
关键词:
摘要: Neuro 2A (N2a) is a mouse neural crest-derived cell line that has been extensively used to study neuronal differentiation, axonal growth and signaling pathways. A convenient characteristic of these cells their ability differentiate into neurons within few days. However, most differentiation methods reported for N2a do not provide information about the types obtained after each treatment. In this study, we evaluated generation dopamine following treatment with number factors known induce differentiation. Our results showed express Nurr-related factor 1 (Nurr1) produce low levels tyrosine hydroxylase (TH) dopamine. Both TH were significantly enhanced in presence dibutyryl cyclic adenosine monophosphate (dbcAMP), as evidenced by Western blot, immunocytochemistry high performance liquid chromatography (HPLC). contrast dbcAMP, other such transforming beta1 (TGF beta 1), bone morphogenetic protein 4 (BMP4), glial cell-derived neurotrophic (GDNF) retinoic acid (RA) did increase expression. Further investigation confirmed effect dbcAMP on production TH-positive was mediated through AMP (cAMP) responsive element binding (CREB) it antagonized RA. Thus, although various treatments can be generate neurons, only formation neurons. Taken together, provided simple reliable method rapid efficient physiological pharmacological assays.
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