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10:46
Mechanical Stimulation-induced Calcium Wave Propagation in Cell Monolayers: The Example of Bovine Corneal Endothelial Cells
Published on: July 16, 2013
Gap junctional intercellular communication in bovine corneal endothelial cells
Priya Gomes1, Sangly P Srinivas, Johan Vereecke
1Laboratory of Physiology, KU Leuven, Campus Gasthuisberg O/N, Box 802, Herestraat 49, B-3000 Leuven, Belgium.
Experimental Eye Research
|August 30, 2006
Summary
This study shows that gap junctions and paracrine mediators both significantly contribute to intercellular communication in bovine corneal endothelial cells. Their interaction is crucial for calcium wave propagation, highlighting complex signaling pathways in non-excitable cells.
Area of Science:
- Cell Biology
- Physiology
Background:
- Intercellular communication (IC) in non-excitable cells relies on gap junctions and paracrine mediators like ATP.
- Bovine corneal endothelial cells (BCEC) are non-excitable and utilize IC for various functions.
Purpose of the Study:
- To investigate the contribution of gap junctions to IC during calcium (Ca2+) wave propagation in BCEC.
- To elucidate the interplay between gap junctional IC (GJIC) and paracrine IC (PIC).
Main Methods:
- Cultured BCEC were stimulated mechanically to induce Ca2+ waves, visualized with Fluo-4.
- Gap junction function was assessed using scrape-loading and fluorescence recovery after photobleaching (FRAP) with hydrophilic dyes.
- Inhibition of GJIC and PIC was achieved using specific blockers (Gap27, apyrases, suramin, Gap26).
Main Results:
- BCEC express Cx43, and functional GJIC was confirmed by dye transfer.
- Blocking gap junctions with Gap27 reduced Ca2+ wave spread by approximately 63%.
- Inhibiting PIC significantly reduced Ca2+ wave spread, with GJIC contributing minimally (approx. 9%) after PIC blockade, suggesting pathway interaction.
Conclusions:
- Functional gap junctions are present in BCEC, mediating significant GJIC.
- Both GJIC and PIC pathways are critical for Ca2+ wave propagation in BCEC.
- Evidence suggests a significant interaction between GJIC and PIC pathways in mediating intercellular communication.
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