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Journal of Experimental Biology, Vol 178, Issue 1 261-267, Copyright © 1993 by Company of Biologists

JOURNAL ARTICLES

ISOLATION OF EPIDERMAL CELL PAIRS FROM AN INSECT, TENEBRIO MOLITOR, FOR DUAL WHOLE-CELL RECORDING OF LARGE-CONDUCTANCE GAP-JUNCTIONAL CHANNELS

D. Churchill and S. Caveney

The segmented insect integument, composed of an epidermal monolayer and its cuticular secretion, is a paradigm for the study of pattern formation during development (Lawrence, 1992). Epidermal activity during development may be coordinated by the transfer of cytoplasmic molecules through cell-cell gap-junctional channels (reviewed in Caveney, 1985). Gap junctions within intact epidermal segments exhibit dynamic changes in their permeability properties during the moult cycle in vivo (Caveney, 1978) and with exposure to the developmental hormone 20-hydroxyecdysone in vitro (Caveney and Blennerhassett, 1980). In addition, the row of epidermal cells at segment borders has distinct permeability properties creating developmental and communication compartments (Warner and Lawrence, 1982; Blennerhassett and Caveney, 1984). This paper documents a method for isolating epidermal cells that are suitable for dual whole-cell voltage-clamp studies of gap-junctional currents. We have identified a large-conductance gap-junctional channel in cell pairs with octanol- reduced junctional currents. This cell model may be useful for examining mechanisms of gap- junctional channel gating during development.




© The Company of Biologists Ltd 1993