Journal
APOPTOSIS
Volume 10, Issue 5, Pages 1019-1029Publisher
SPRINGER
DOI: 10.1007/s10495-005-0776-8
Keywords
annexin V; apoptosis; BHK cells; bystander cell death; caspase activity; connexin32; cytochrome C; gap junction; intercellular communication; transfection; TUNEL staining
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Funding
- NEI NIH HHS [EY 06516, EY 01792] Funding Source: Medline
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Programmed cell death (apoptosis) occurs both during normal development and as a result of various pathological conditions. An in vitro system was used to explore the transmission of death signals from apoptotic cells to cells with which they were coupled via gap junctions. Confluent cultures of baby hamster kidney (BHK) cells, stably transfected with the gap-junctional protein connexin32, were scrape loaded with cytochrome C (cyC), a mitochondria-derived apoptotic agent, to introduce the protein into cells injured by the cut. The cultures were subsequently analyzed for the presence of activated caspases, the distribution of TUNEL staining, and the binding of annexin V. Although cyC is too large to traverse the gap junctional channel, each of the assays revealed that apoptosis had spread from dying cells at the margin of the scrape to otherwise healthy neighboring cells to which they were coupled. This 'bystander effect' was significantly reduced in the presence of agents that block gap junctional intercellular communication.
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