The effect of alternative neuronal differentiation pathways on PC12 cell adhesion and neurite alignment to nanogratings.
During development and regeneration of the mammalian nervous system, directional signals guide differentiating neurons toward their targets. Soluble neurotrophic molecules encode for preferential direction over long distances while the local topography is read by cells in a process requiring the establishment of focal adhesions. The mutual interaction between overlapping molecular and topographical signals introduces an additional level of control to this picture. The role of the substrate topography was demonstrated exploiting nanotechnologies to generate biomimetic scaffolds that control both the polarity of differentiating neurons and the alignment of their neurites. Here PC12 cells contacting nanogratings made of copolymer 2-norbornene ethylene (COC), were alternatively stimulated with Nerve Growth Factor, Forskolin, and 8-(4-chloro-phenylthio)-2'-O-methyladenosine-3',5'-cyclic (8CPT-2Me-cAMP) or with a combination of them. Topographical guidance was differently modulated by the alternative stimulation protocols tested. Forskolin stimulation reduced the efficiency of neurite alignment to the nanogratings. This effect was linked to the inhibition of focal adhesion maturation. Modulation of neurite alignment and focal adhesion maturation upon Forskolin stimulation depended on the activation of the MEK/ERK signaling but were PkA independent. Altogether, our results demonstrate that topographical guidance in PC12 cells is modulated by the activation of alternative neuronal differentiation pathways.
Ferrari
Aldo
A
NEST, Scuola Normale Superiore and CNR-INFM, Piazza San Silvestro, 12 I-56126 Pisa, Italy. aldo.ferrari@ltnt.iet.mavt.ethz.ch
Faraci
Paolo
P
Cecchini
Marco
M
Beltram
Fabio
F
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D022001:Focal Adhesions / Q000187:drug effects / Q000201:enzymology; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D049329:Nanostructures / Q000737:chemistry*; D016501:Neurites / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D016716:PC12 Cells; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051381:Rats
D047428:Protein Kinase Inhibitors; D017868:Cyclic AMP-Dependent Protein Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D020929:Mitogen-Activated Protein Kinase Kinases