| Transfection of neonatal rat Schwann cells with SV-40 large T antigen gene under control of the metallothionein promoter. | |
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MedLine Citation:
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PMID: 2824529 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Secondary cultures of Schwann cells were transfected with a plasmid containing the SV-40 T antigen gene expressed under the control of the mouse metallothionein-I promoter. We used the calcium phosphate method for transfection and obtained a transfection efficiency of 0.01%. The colonies were cloned by limited dilution, and these cloned cell lines were carried in medium containing zinc chloride (100 microM). One cloned cell line, which has now been carried for 180 doublings, appears to have a transformed phenotype with a doubling time of 20 h. These cells express SV-40 T antigen while maintaining established Schwann cell properties (positive staining for 217c, Ran-2, A5E3, glial fibrillary acidic protein, presence of 2',3'-cyclic nucleotide phosphohydrolase [CNPase] activity, and the ability to synthesize sulfogalactosylceramide and mRNA for the myelin protein, P0). Removal of zinc chloride from the medium resulted in reduced expression of T antigen and a change in the appearance of the cells to a more bipolar shape, although they still did not exhibit contact inhibition and maintained a doubling time of 20 h. These cells now became Ran-2-negative and showed increases in CNPase activity and in their ability to synthesize sulfogalactosylceramide. The amount of P0 mRNA remained unchanged. Transfected Schwann cells, however, stopped dividing when they contacted either basal lamina or neurites and became bipolar in appearance. The Schwann cells in contact with the neurites then extended processes to wrap around bundles of neurites. Transfection with the SV-40 T antigen gene therefore provides a method for obtaining Schwann cell lines that continue to express properties associated with untransfected cells in culture and may be used to study axon-Schwann cell interaction. |
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Authors:
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G I Tennekoon; J Yoshino; K W Peden; J Bigbee; J L Rutkowski; Y Kishimoto; G H DeVries; G M McKhann |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: The Journal of cell biology Volume: 105 ISSN: 0021-9525 ISO Abbreviation: J. Cell Biol. Publication Date: 1987 Nov |
Date Detail:
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Created Date: 1988-01-07 Completed Date: 1988-01-07 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 0375356 Medline TA: J Cell Biol Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 2315-25 Citation Subset: IM |
Affiliation:
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Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205. |
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Animals, Newborn Antigens, Polyomavirus Transforming / genetics* Cell Transformation, Viral* Cells, Cultured Ganglia, Spinal / cytology, ultrastructure Genes* Genes, Viral* Metallothionein / genetics* Microscopy, Electron Promoter Regions, Genetic* Rats Rats, Inbred Strains Schwann Cells / cytology, immunology*, ultrastructure Simian virus 40 / genetics* |
| Grant Support | |
ID/Acronym/Agency:
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1PO122845//PHS HHS; NS-21700/NS/NINDS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Antigens, Polyomavirus Transforming; 9038-94-2/Metallothionein |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
| Full Text | |
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Journal Information Journal ID (nlm-ta): J Cell Biol ISSN: 0021-9525 ISSN: 1540-8140 Publisher: The Rockefeller University Press |
Article Information Download PDF  Print publication date: Day: 1 Month: 11 Year: 1987 Volume: 105 Issue: 5 First Page: 2315 Last Page: 2325 ID: 2114872 Publisher Id: 88059263 PubMed Id: 2824529 |
| Transfection of neonatal rat Schwann cells with SV-40 large T antigen gene under control of the metallothionein promoter | |
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