Gap junction intercellular communication: a microinjection investigation of fibroblast and epithelial cell lines

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Pahlka, Raymond Benton
Texas A&M University
health physics.; Major health physics.
thesis / dissertation description
The objectives of this research were threefold. The first objective was to develop a protocol for unbiased microinjection of the fluorescent dye Lucifer Yellow to normal fibroblast and epithelial cell lines. I determined the optimal equipment parameters as well as optimal cell conditions for effective, repeatable studies using the microinjection protocol. The second objective was to determine whether or not the AG1522 cell line exhibited gap junction intercellular communication (GJIC) through microinjection of the fluorescent dye Lucifer Yellow. A resolution was reached and these studies indicate that the AG1522 cell line does not exhibit GJIC on the basis of Lucifer Yellow microinjection. The third objective entailed determining to what extent neighboring Clone 9 cell colonies in contact with each other affected GJIC. These studies indicate a hierarchy of gap-junction communication ranging from negligible communication in single cell colony-to-colony boundaries to maximal communication in confluent cell monolayers. To better characterize the communication in these Clone 9 cells, I compared experimental data of dye concentrations in confluent monolayers as functions of time and distance away from the injected cell to two different analytical solutions of the diffusion equation. I found that the experimental data correlated well with one solution and thus calculated a corresponding diffusion coefficient of 68 []m²/sec. Investigations and observations of some experimental anomalies were discussed and future investigations concerning the effects of ionizing radiation on gap-junction expression using various assays of GJIC were proposed.