Document Detail


Isolation and characterization of Chinese hamster cells defective in cell-cell coupling via gap junctions.
MedLine Citation:
PMID:  6840215     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Chinese hamster Wg3-h-o cells which were descended from DON cells have been mutagenized and selected for derivatives defective in metabolic cooperation via gap junctions (i.e., mec-). The selection protocol included four consecutive cycles of cocultivating mutagenized cells, deficient in hypoxanthine phosphoribosyltransferase (HPRT) and wild-type cells in the presence of thioguanine (cf Slack, C, Morgan, R H M & Hooper, M L, Exp cell res 117 (1978) 195-205) [8]. We carried out the last two selection cycles in the presence of 1 mM dibutyryl cyclic adenosine monophosphate (db-cAMP). The isolated Chinese hamster CI-4 cells which expressed the mec- phenotype most stringently showed the following characteristics: 1. In standard culture medium no cell-cell coupling was detected among CI-4 cells when assayed by injections of the fluorescent dye Lucifer yellow or by electrical measurements. Between 73 and 100% of the mec+ parental cells were coupled under these conditions. Up to 14% positive contacts were found between CI-4 cells and Chinese hamster Don cells (mec+). Confluent CI-4 cells grown in the presence of 1 mM db-cAMP showed 9% coupled cells. 2. No gap junction plaques were found on electron micrographs of freeze-fractured, confluent CI-4 cells. The mec+ parental cells showed small gap junction plaques (0.013% of the total cell surface analyzed). 3. CI-4 cells exhibited 16% positive contacts and the parental Wg3-h-o cells showed 92% positive contacts in autoradiographic measurements of metabolic cooperation with DON cells. On an extracellular matrix, prepared from normal embryonic fibroblasts, metabolic cooperation between CI-4 and DON cells was autoradiographically measured to be 68%. Other cells of spontaneous mec- phenotype (for example mouse L cells or human fibrosarcoma HT1080 cells) also appeared to exhibit increased metabolic cooperation when grown on an extracellular matrix and assayed by autoradiographic measurements. When tested by Lucifer yellow injections, however, only very few positive contacts were found for CI-4/DON cell pairs and no positive contacts were found among mouse L cells grown on an extracellular matrix. 4. The mec- defect in the genome of CI-4 cells was cured in somatic cell hybrids with mouse embryonic fibroblasts or with mouse embryonal carcinoma cells. The results of isozyme and karyotype studies of mec-, as well as mec+ somatic cell hybrids suggest that mouse chromosome 16 may be involved in complementation of the mec- defect.
Authors:
K Willecke; D Müller; P M Drüge; U Frixen; R Schäfer; R Dermietzel; D Hülser
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Experimental cell research     Volume:  144     ISSN:  0014-4827     ISO Abbreviation:  Exp. Cell Res.     Publication Date:  1983 Mar 
Date Detail:
Created Date:  1983-06-10     Completed Date:  1983-06-10     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0373226     Medline TA:  Exp Cell Res     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  95-113     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Autoradiography
Cell Communication*
Cell Line
Cell Separation
Chromosomes / physiology
Cricetinae
Electrophysiology
Genes*
Genetic Complementation Test
Hybrid Cells / physiology
Intercellular Junctions / physiology*,  ultrastructure
Isoenzymes / analysis
Isoquinolines
Karyotyping
Mutation
Chemical
Reg. No./Substance:
0/Isoenzymes; 0/Isoquinolines; 77944-88-8/lucifer yellow

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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