| Rat-1 fibroblasts engineered with GAD65 and GAD67 cDNAs in retroviral vectors produce and release GABA. | |
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MedLine Citation:
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PMID: 8336154 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We have used a retroviral cDNA expression system to drive the expression of the different forms of glutamic acid decarboxylase (GAD65, GAD67, or both). Individual clones of engineered Rat-1 cells make the appropriate GAD mRNAs and GAD polypeptides, show GAD enzymatic activity, and make GABA. Clones expressing GAD65 had higher enzymatic activity than those expressing GAD67. As is the case for brain GADs and for GADs produced in engineered bacteria, the enzymatic activity of GAD65 is more responsive to added pyridoxal phosphate than that of GAD67. Immunostaining for both GADs is scattered throughout the cytoplasm. GAD65 immunostaining is less homogeneous than that of GAD67 and also appears to be associated with the surfaces of large vesicle-like structures. Cells expressing GAD65 and GAD67 showed similar immunostaining patterns with anti-GABA antibodies and contained substantial amounts of GABA (ranging from 7 to 18 pmol of GABA/10(6) cells), which was roughly proportional to their levels of GAD activity. GABA is released from the engineered cells into the surrounding medium under resting conditions, suggesting that cells programmed with GAD cDNAs might serve as effective sources of GABA in cell transplantation experiments. |
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Authors:
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C Ruppert; A Sandrasagra; B Anton; C Evans; E S Schweitzer; A J Tobin |
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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: Journal of neurochemistry Volume: 61 ISSN: 0022-3042 ISO Abbreviation: J. Neurochem. Publication Date: 1993 Aug |
Date Detail:
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Created Date: 1993-08-24 Completed Date: 1993-08-24 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 2985190R Medline TA: J Neurochem Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 768-71 Citation Subset: IM |
Affiliation:
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Department of Biology, University of California, Los Angeles 90024-1606. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Line DNA / genetics* Fibroblasts / metabolism* Gene Expression Genetic Engineering Genetic Vectors Glutamate Decarboxylase / analysis, genetics* Immunoenzyme Techniques Rats Retroviridae / genetics* Transfection* gamma-Aminobutyric Acid / analysis, biosynthesis*, secretion |
| Grant Support | |
ID/Acronym/Agency:
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NS22256/NS/NINDS NIH HHS; NS23084/NS/NINDS NIH HHS |
| Chemical | |
Reg. No./Substance:
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56-12-2/gamma-Aminobutyric Acid; 9007-49-2/DNA; EC 4.1.1.15/Glutamate Decarboxylase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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