title: Functional and morphological preservation of adult ventricular myocytes in culture by sub-micromolar cytochalasin D supplement.
abstract: In cardiac myocytes, cytochalasin D (CytoD) was reported to act as an actin disruptor and mechanical uncoupler. Using confocal and super-resolution STED microscopy, we show that CytoD preserves the actin filament architecture of adult rat ventricular myocytes in culture. Five hundred nanomolar CytoD was the optimal concentration to achieve both preservation of the T-tubular structure during culture periods of 3 days and conservation of major functional characteristics such as action potentials, calcium transients and, importantly, the contractile properties of single myocytes. Therefore, we conclude that the addition of CytoD to the culture of adult cardiac myocytes can indeed be used to generate a solid single-cell model that preserves both morphology and function of freshly isolated cells. Moreover, we reveal a putative link between cytoskeletal and T-tubular remodeling. In the absence of CytoD, we observed a loss of T-tubules that led to significant dyssynchronous Ca(2+)-induced Ca(2+) release (CICR), while in the presence of 0.5 μM CytoD, T-tubules and homogeneous CICR were majorly preserved. Such data suggested a possible link between the actin cytoskeleton, T-tubules and synchronous, reliable excitation-contraction-coupling. Thus, T-tubular re-organization in cell culture sheds some additional light onto similar processes found during many cardiac diseases and might link cytoskeletal alterations to changes in subcellular Ca(2+) signaling revealed under such pathophysiological conditions.
authors.affiliation: Institute for Molecular Cell Biology, Medical Faculty, Building 61, Saarland University, 66421 Homburg/Saar, Germany.
authors.forename: Qinghai
authors.initials: Q
authors.lastname: Tian
authors.forename: Sara
authors.initials: S
authors.lastname: Pahlavan
authors.forename: Katharina
authors.initials: K
authors.lastname: Oleinikow
authors.forename: Jennifer
authors.initials: J
authors.lastname: Jung
authors.forename: Sandra
authors.initials: S
authors.lastname: Ruppenthal
authors.forename: Anke
authors.initials: A
authors.lastname: Scholz
authors.forename: Christian
authors.initials: C
authors.lastname: Schumann
authors.forename: Annette
authors.initials: A
authors.lastname: Kraegeloh
authors.forename: Martin
authors.initials: M
authors.lastname: Oberhofer
authors.forename: Peter
authors.initials: P
authors.lastname: Lipp
authors.forename: Lars
authors.initials: L
authors.lastname: Kaestner
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects; D015638:Cytochalasin D / Q000378:metabolism / Q000494:pharmacology*; D006352:Heart Ventricles / Q000166:cytology / Q000378:metabolism; D008297:Male; D009200:Myocardial Contraction / Q000187:drug effects; D032383:Myocytes, Cardiac / Q000000981:diagnostic imaging* / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D014463:Ultrasonography
chemical_list: D015638:Cytochalasin D; D002118:Calcium