Generation of myocyte agonal Ca²⁺ waves and contraction bands in perfused rat hearts following irreversible membrane permeabilisation.

ABSTRACT: Although irreversible cardiomyocyte injury provokes intracellular Ca²⁺ ([Ca²⁺]_i) overload, the underlying dynamics of this response and its effects on cellular morphology remain unknown. We therefore visualised rapid-scanning confocal fluo4-[Ca²⁺]_i dynamics and morphology of cardiomyocytes in Langendorff-perfused rat hearts following saponin-membrane permeabilisation. Our data demonstrate that 0.4% saponin-treated myocytes immediately exhibited high-frequency Ca²⁺ waves (131.3 waves/min/cell) with asynchronous, oscillatory contractions having a mean propagation velocity of 117.8 μm/s. These waves slowly decreased in frequency, developed a prolonged decay phase, and disappeared in 10 min resulting in high-static, fluo4-fluorescence intensity. The myocytes showing these waves displayed contraction bands, i.e., band-like actin-fibre aggregates with disruption of sarcomeric α-actinin. The contraction bands were not attenuated by the abolition of Ca²⁺ waves under pretreatment with ryanodine plus thapsigargin, but were partially attenuated by the calpain inhibitor MDL28170, while mechanical arrest of the myocytes by 2,3-butanedione monoxime completely attenuated contraction-band formation. The depletion of adenosine 5'-triphosphate by the mitochondrial electron uncoupler carbonyl cyanide 4-trifluoromethoxy phenylhydrazone also attenuated Ca²⁺ waves and contraction bands. Overall, saponin-induced myocyte [Ca²⁺]_i overload provokes agonal Ca²⁺ waves and contraction bands. Contraction bands are not the direct consequence of the waves but are caused by cross-bridge interactions of the myocytes under calpain-mediated proteolysis.

SUBMITTER: Morishita Y 

PROVIDER: S-EPMC9842683 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature