Direct isolation and analysis of endogenous transjunctional ADP from Cx43 transfected C6 glioma cells

Gary S. Goldberg, Paul D. Lampe, David Sheedy, Carleton C. Stewart, Bruce J. Nicholson, Christian C.G. Naus

Research output: Contribution to journalArticle

61 Scopus citations

Abstract

Gap junctional communication has been implicated in numerous cellular processes. However, the repertoire of specific transjunctional substances which mediate these processes remains relatively unexplored. A few selected secondary messengers have been identified, at least indirectly (e.g., cAMP and IP3) and phenotypic complementation experiments have indicated that gap junctions enable communicating cells to distribute nucleotide pools as a shared resource. The latter would include high energy compounds such as ADP and ATP, allowing cells to share energy resources. We have utilized a nonbiased process to directly capture, identify, and quantify transjunctional compounds from C6 glioma cells, the transformed phenotype of which has been ameliorated by transfection with connexin43 (Cx43). This technique involves the direct isolation, identification, and quantitation of radioactive transjunctional molecules that travel from metabolically labeled 'donor' cells to 'receiver' cells. This report demonstrates that ADP and/or ATP represents over 6% of the transjunctional material derived from glucose in Cx43-transfected C6 glioma cells. Furthermore, equilibration of these high energy metabolites among first order neighbors is shown to occur in less than 20 min of communication.

Original languageEnglish (US)
Pages (from-to)82-92
Number of pages11
JournalExperimental Cell Research
Volume239
Issue number1
DOIs
StatePublished - Feb 25 1998
Externally publishedYes

ASJC Scopus subject areas

  • Cell Biology

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    Goldberg, G. S., Lampe, P. D., Sheedy, D., Stewart, C. C., Nicholson, B. J., & Naus, C. C. G. (1998). Direct isolation and analysis of endogenous transjunctional ADP from Cx43 transfected C6 glioma cells. Experimental Cell Research, 239(1), 82-92. https://doi.org/10.1006/excr.1997.3872