tretinoin and 8-bromoguanosino-3--5--cyclic-monophosphorothioate

Developmental studies in both vertebrates and invertebrates implicate an involvement of nitric oxide (NO) signaling in cell proliferation, neuronal motility, and synaptic maturation. However, it is unknown whether NO plays a role in the development of the human nervous system. We used a model of human neuronal precursor cells from a well-characterized teratocarcinoma cell line (NT2). The precursor cells proliferate during retinoic acid treatment as spherical aggregate culture that stains for nestin and betaIII-tubulin. Cells migrate out of the aggregates to acquire fully differentiated neuronal phenotypes. The cells express neuronal nitric oxide synthase and soluble guanylyl cyclase (sGC), an enzyme that synthesizes cGMP upon activation by NO. The migration of the neuronal precursor cell is blocked by the use of nNOS, sGC, and protein kinase G (PKG) inhibitors. Inhibition of sGC can be rescued by a membrane permeable analog of cGMP. In gain of function experiments the application of a NO donor and cGMP analog facilitate cell migration. Our results from the differentiating NT2 model neurons point towards a vital role of the NO/cGMP/PKG signaling cascade as positive regulator of cell migration in the developing human brain.

Topics: Cell Differentiation; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Cyclic GMP; Dose-Response Relationship, Drug; Enzyme Inhibitors; Gene Expression Regulation; Humans; Models, Biological; Neoplastic Stem Cells; Neurons; Nitric Oxide; Nitric Oxide Donors; Nitric Oxide Synthase Type I; Nitroso Compounds; Signal Transduction; Thionucleotides; Time Factors; Tretinoin; Tubulin