From transcription profile to expression: the signaling repertoire of the SK-N-MC neuroepithelioma cell-line
| Autor: | Norman Koglin, Hans-Jürgen Thiesen, Dirk Koczan, Karin Mörl, Annette G. Beck-Sickinger, Jürgen E. Bader, Cornelia M. Deckert, Franka Pluder |
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| Rok vydání: | 2005 |
| Předmět: |
Receptors
Neuropeptide Transcription Genetic Blotting Western Immunoblotting Biology Biochemistry RNA Complementary Receptors G-Protein-Coupled Inhibitory Concentration 50 Transcription (biology) Orexin Receptors Cell Line Tumor mental disorders Gene expression Humans Neuroectodermal Tumors Primitive Peripheral Receptor Molecular Biology Oligonucleotide Array Sequence Analysis Binding Sites Reverse Transcriptase Polymerase Chain Reaction Cell Membrane Cell Biology Neuropeptide Y receptor Molecular biology Orexin receptor Orexin Receptors Neuropeptide Y Proteome Gene chip analysis RNA Electrophoresis Polyacrylamide Gel Peptides Protein Binding Signal Transduction |
| Zdroj: | Journal of receptor and signal transduction research. 24(4) |
| ISSN: | 1079-9893 |
| Popis: | SK-N-MC neuroepithelioma cells are routinely cultured and widely used as a model system in biochemical and pharmacological experiments. To clarify the gene expression patterns of SK-N-MC cells with respect to G protein-coupled receptors and signaling network components, we describe in this report the transcription profile of the cell line. Following the traditional pathway from genome to proteome, selected examples are further examined at the level of protein expression and by functional assays. cRNA targets derived from total RNA extracts were hybridized to Affymetrix Human Genome U133A GeneChip arrays, and the data were analyzed and grouped according to functional aspects. Results obtained for neuropeptide Y (NPY) Y1, Y5, and orexin Ox1 receptors were confirmed by RT-PCR. It is surprising that we found the presence of both NPY receptor subtypes and the absence of the orexin receptor at the mRNA level. Receptor-binding experiments confirmed NPY binding of the Y1 receptor in the nanomolar range but gave no evidence for high expression levels of Y5 receptor subtypes on the cell surface. Protein expression was assayed with immunoblots by using antibodies directed against selected Galpha protein subunits. The presence of at least Galphas, Galphai3, and Galphai2 subunits was indicated. |
| Databáze: | OpenAIRE |
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