Application of a Rapid Method (Targeted Display) for the Identification of Differentially Expressed mRNAs Following NGF-Induced Neuronal Differentiation in PC12 Cells
| Autor: | Julian F. Burke, Alastair J.H. Brown, C. Hutchings, Lynne V. Mayne |
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| Rok vydání: | 1999 |
| Předmět: |
DNA
Complementary Transcription Genetic Cellular differentiation Protein Serine-Threonine Kinases Biology Transfection PC12 Cells Polymerase Chain Reaction Gene product Mice Cellular and Molecular Neuroscience Sequence Homology Nucleic Acid Gene expression Animals Nerve Growth Factors RNA Messenger RNA Neoplasm Northern blot Molecular Biology Electrophoresis Agar Gel Regulation of gene expression Gene Expression Regulation Developmental Cell Differentiation Cell Biology Molecular biology Rats Fibroblast Growth Factors Gene Expression Regulation Neoplastic Nerve growth factor nervous system Cell culture |
| Zdroj: | Molecular and Cellular Neuroscience. 13:119-130 |
| ISSN: | 1044-7431 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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