Application of a rapid method (targeted display) for the identification of differentially expressed mRNAs following NGF-induced neuronal differentiation in PC12 cells

Mol Cell Neurosci. 1999 Feb;13(2):119-30. doi: 10.1006/mcne.1999.0736.

Abstract

Nerve growth factor (NGF)-induced differentiation of the rat pheochromocytoma, PC12, cell line presents a model system for the study of early gene expression changes involved in neuronal differentiation. Rapid alterations in mRNA expression patterns were investigated in PC12 cells following exposure to NGF using a set of statistically designed primers that exhibit coding-strand bias, and the products were analyzed on agarose gels. This simple and rapid method (targeted display) generated reproducible expression profiles, indicating a complex pattern of gene regulation, and resulted in the identification of a number of NGF-regulated transcripts. Thirty-two of these were selected at random and sequenced, revealing 19 known and 13 novel genes (or ESTs). Northern blot analysis and RT-PCR confirmed the differential regulation of 22 genes (16 known, 6 novel) and demonstrated 1 false positive result. Antisense application of one isolated gene product, the serine/threonine kinase MARK1, prevented neuronal differentiation in transiently transfected PC12 cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / drug effects
  • DNA, Complementary / genetics
  • Electrophoresis, Agar Gel / methods
  • Fibroblast Growth Factors / pharmacology
  • Gene Expression Regulation, Developmental / drug effects*
  • Gene Expression Regulation, Neoplastic / drug effects*
  • Mice
  • Nerve Growth Factors / pharmacology*
  • PC12 Cells / drug effects*
  • PC12 Cells / metabolism
  • Polymerase Chain Reaction / methods*
  • Protein-Serine-Threonine Kinases / genetics
  • Protein-Serine-Threonine Kinases / physiology
  • RNA, Messenger / biosynthesis*
  • RNA, Neoplasm / biosynthesis
  • Rats
  • Sequence Homology, Nucleic Acid
  • Transcription, Genetic
  • Transfection

Substances

  • DNA, Complementary
  • Nerve Growth Factors
  • RNA, Messenger
  • RNA, Neoplasm
  • Fibroblast Growth Factors
  • Mark1 protein, rat
  • Protein-Serine-Threonine Kinases