Priming of PC12 cells for semiquantitative microinjection studies involving Ras.
Nerve growth factor and activated Ras can induce differentiation of rat pheochromocytoma cells (PC12 cells) [Greene and Tischler (1976) Proc. Natl. Acad. Sci. USA 73, 2424-2428] from a chromaffin cell-like morphology into one that resembles sympathetic neurones. We developed a special treatment of PC12 cells which apparently synchronizes these cells such that they are more useful for semi-quantitative microinjection studies for signal transduction pathways. This treatment leads to a faster and more reproducible differentiation which faithfully reproduces the involvement of Ras in the process and allows a comparison of the biological activity of different Ras mutants. It shows that G12V and Q61L oncogenic mutants are not equally potent in inducing differentiation. Partial loss-of-function mutations T35S, E37G and Y40C are inactive and even a triple combination of these does not restore full biological activity.
Schmidt
G
G
Max-Planck-Institut für molekulare Physiologie, Otto-Hahn-Str. 11, 44227, Dortmund, Germany.
Wittinghofer
A
A
D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D003609:Dactinomycin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D008845:Microinjections; D009154:Mutation / Q000235:genetics; D020932:Nerve Growth Factor / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D016501:Neurites / Q000187:drug effects / Q000378:metabolism; D015689:Oncogene Protein p21(ras) / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D016716:PC12 Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects; D018631:ras Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*
D003609:Dactinomycin; D020932:Nerve Growth Factor; D015689:Oncogene Protein p21(ras); D018631:ras Proteins