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Priming of PC12 cells for semiquantitative microinjection studies involving Ras
Author(s) -
Schmidt Gudula,
Wittinghofer Alfred
Publication year - 2000
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(00)01602-1
Subject(s) - microinjection , priming (agriculture) , chemistry , microbiology and biotechnology , andrology , biology , genetics , medicine , botany , germination
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 synchronises 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.