Transfection with trk restores "slow" NGF binding, efficient NGF uptake, and multiple NGF responses to NGF-nonresponsive PC 12 cell mutants

David M. Loeb, Lloyd A. Greene

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

NGF binds to and activates the protein tyrosine kinase gp140prototrk. Expression of this receptor is required for at least some responses to NGF. Three outstanding issues are addressed in the present work. First, we determined whether expression of gp140prototrk is required for all neuronal NGF responses. Second, we examined the role of gp140prototrk in NGF binding and internalization. Third, we addressed the utility of NGF-nonresponsive PC12nnr5 cells for study of the NGF mechanism. In contrast to wild-type PC12 cells, PC12nnr5 cells do not express endogenous gp140prototrk. We therefore asked whether they possess other defects that compromise NGF signaling pathways. To answer these questions, we transfected PC12nnr5 cells with a cDNA encoding full-length human gp140prototrk and isolated cell lines permanently expressing the receptor. Introduction of trk rescued all of the many and varied NGF responses assessed, including enhanced protein tyrosine phosphorylation, induction of immediate-early and neural-specific genes, neurite outgrowth and regeneration, maintenance of survival in serum-free medium, and stimulation of AChE activity. In contrast to PC12nnr5 cells, the trk-transfected lines also bind and internalize NGF with wild-type PC12 cell characteristics. These findings indicate that gp140prototrk is required for many, if not all, responses of neuronal cells to NGF and is necessary for proper NGF binding and internalization. Additionally, as no signaling defect other than the absence of trk expression was revealed in PC12nnr5 cells, this work supports the utility of this line for genetic dissection of the NGF mechanism of action.

Original languageEnglish (US)
Pages (from-to)2919-2929
Number of pages11
JournalJournal of Neuroscience
Volume13
Issue number7
StatePublished - Dec 1 1993
Externally publishedYes

Fingerprint

Nerve Growth Factor
Transfection
PC12 Cells
Serum-Free Culture Media
Protein-Tyrosine Kinases
Tyrosine
Dissection
Regeneration
Complementary DNA
Maintenance
Phosphorylation

Keywords

  • NGF
  • NGF binding
  • NGF internalization
  • NGF receptor
  • PC12 cells
  • PC12nnr cells
  • Signal transduction
  • Trk
  • Tyrosine kinase

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Transfection with trk restores "slow" NGF binding, efficient NGF uptake, and multiple NGF responses to NGF-nonresponsive PC 12 cell mutants. / Loeb, David M.; Greene, Lloyd A.

In: Journal of Neuroscience, Vol. 13, No. 7, 01.12.1993, p. 2919-2929.

Research output: Contribution to journalArticle

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