Quantitation of the growth-associated protein B-50/GAP-43 and neurite outgrowth in PC12 cells.
A combined assay to measure neurite outgrowth and B-50/GAP-43 levels in PC12 cells is reported. During NGF-induced neuritogenesis, B-50/GAP-43 expression was monitored by enzyme-linked immunosorbent assay (ELISA). Neurite outgrowth was quantified at the same time by the use of video image analysis. Sensitivity and reliability of the methods are shown with a dose-response and time curve of beta-NGF-induced neuritogenesis. A linear increase in total length of neurites was induced by concentrations of beta-NGF greater than or equal to 5 ng/ml and was accompanied by a linear increase in the amount of B-50/GAP-43. The combined methods presented here can conveniently and reliably establish subtle changes in neurite outgrowth and intracellular protein contents.
Jap Tjoen San
E R
ER
Division of Molecular Neurobiology, Rudolf Magnus Institute, University of Utrecht, The Netherlands.
Schmidt-Michels
M H
MH
Spruijt
B M
BM
Oestreicher
A B
AB
Schotman
P
P
Gispen
W H
WH
D000310:Adrenal Gland Neoplasms / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D001369:Axons / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D019922:GAP-43 Protein; D006133:Growth Substances / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009994:Osmolar Concentration; D010673:Pheochromocytoma / Q000378:metabolism* / Q000648:ultrastructure; D013997:Time Factors; D014407:Tumor Cells, Cultured
D019922:GAP-43 Protein; D006133:Growth Substances; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins