Key result
Imposing a small increment in free calcium activity (14 to 17 microM) during dissociation of adult rat hearts yields functionally intact myocyte pairs with strong electrical coupling.
A small increment in free calcium during dissociation of adult rat hearts yields functionally intact myocyte pairs with stronger electrical coupling than newly formed pairs.
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Supports refined myocyte-pair isolation for coupling studies; leaves open human translation.
The dissociation of adult rat heart into individual, functionally intact, calcium-tolerant myocytes requires precise manipulation of extracellular calcium levels. Dissociation of intercellular connections is achieved by lowering extracellular calcium to micromolar levels for a short period. By imposing a very small increment in free calcium activity (from 14 to 17 microM) during this period, we achieve a significant yield of functionally intact pairs of myocytes still joined at the intercalated disc. We obtain fewer intact cells, but many of these are paired end to end. These findings permit us to describe some structural characteristics of intercellular connections between cardiac cells and to report unambiguous measurements of electrotonic coupling and dye transfer between rat cardiac cell pairs. We find that the strength of electrical coupling between cells isolated as pairs with intact junctional contacts is much greater than that measured between cell pairs that have formed new junctional contacts.
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Wittenberg et al. (1986) studied Adult rat heart myocytes. Increment in free calcium activity was evaluated on Yield of functionally intact pairs of myocytes and strength of electrical coupling. Imposing a small increment in free calcium activity (14 to 17 microM) during dissociation of adult rat hearts yields functionally intact myocyte pairs with strong electrical coupling.
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