Immunocytochemical analysis of the regeneration of myofibrils in long-term cultures of adult cardiomyocytes of the rat

Eppenberger, Monika E.; I, Hauser; Baechi, Thomas; Schaub, Marcus C.; Brünner, U; Dechesne, Claude J.; Eppenberger, Hans M.

doi:10.1016/0012-1606(88)90408-3

Cited by 87 publications

(67 citation statements)

References 56 publications

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“…Differential isoprotein distribution in the regenerating ARCS has already been observed. The a-MHC appears to localize primarily in the sarcomeres, whereas the f3-MHC is found in both myofibrillar and stress fiber structures (Eppenberger et al, 1988). Similarly, it was very recently reported that the a-vascular actin is preferentially found in the nonsarcomeric cytoskeleton, whereas the sarcomeric actin isoforms localize in the myofibrils (Eppenberger-Eberhardt et al, 1990).…”

Section: Mutagenesismentioning

confidence: 84%

“…In regenerating ARCS, two distinct types of actin-myosin cytoskeletons coexist (Eppenberger et al, 1988;Eppenberger-Eberhardt et al, 1990). When expressed in these cells, exogenous skeletal LC-1 f and LC-3f bind preferentially to the sarcomeric MHC and therefore are found almost exclusively in myofibrils, often found in the perinuclear region.…”

Section: Discussionmentioning

confidence: 99%

“…The following experiments were carried out in ARCS. These adult cells have been shown to undergo a cycle of dedifferentiation and redifferentiation when kept in long-term culture (Eppenberger et al, 1988). During this process, after a period of extensive myofibril disassembly, the ARCS regenerate from a mainly "stress fiber scaffold" cytoskeleton to a well-organized myofibrillar cytoskeleton.…”

Section: Mutagenesismentioning

confidence: 99%

“…In most regenerating ARCS the de novo building up of sarcomeres appears to be concentrated in a perinuclear region (Figures 3b and 3f), but two other prominent sites are revealed by incorporation of the LC-lfTl 1 and LC3f-Tl l, namely the intercalated disk region between adjacent cells (Figure 3f; arrow) and also some discrete "patches" (Figure 3b; arrowheads). These "patches" likely represent presarcomeric structures in physical continuity with the stress fiber-like scaffold (Eppenberger et al, 1988) and are sometimes found quite distant to the perinuclear region (Figure 3b; arrowheads). Interestingly, the anti-tag staining also reveals some small periodic structures ( Figure 3D; arrowheads) very similar to staining patterns described by others after microinjection of rhodamine-labeled LCs in chickembryonic myotubes (Mittal et al, 1987).…”

Section: Mutagenesismentioning

confidence: 99%

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Intracompartmental sorting of essential myosin light chains: Molecular dissection and in vivo monitoring by epitope tagging

Soldati¹,

Perriard

1991

Cell

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Section: Mutagenesismentioning

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Section: Mutagenesismentioning

confidence: 99%

Section: Mutagenesismentioning

confidence: 99%

See 2 more Smart Citations

Intracompartmental sorting of essential myosin light chains: Molecular dissection and in vivo monitoring by epitope tagging

Soldati¹,

Perriard

1991

Cell

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“…In cell culture medium containing serum ARVM undergo a process of de and redifferentiation, reestablish cell-cell contacts, regenerate myofibrils and resume spontaneous contractions (Eppenberger and Zuppinger 1999;Eppenberger et al 1988) (Fig. 1a-d).…”

Section: Structure and Function Of Arvm After Isolation And In Long-tmentioning

confidence: 99%

The role of cell death and myofibrillar damage in contractile dysfunction of long-term cultured adult cardiomyocytes exposed to doxorubicin

et al. 2009

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In failing hearts cardiomyocytes undergo alterations in cytoskeleton structure, contractility and viability. It is not known presently, how stress-induced changes of myofibrils correlate with markers for cell death and contractile function in cardiomyocytes. Therefore, we have studied the progression of contractile dysfunction, myofibrillar damage and cell death in cultured adult cardiomyocytes exposed to the cancer therapy doxorubicin. We demonstrate, that long-term cultured adult cardiomyocytes, a wellestablished model for the study of myofibrillar structure and effects of growth factors, can also be used to assess contractility and calcium handling. Adult rat ventricular myocytes (ARVM) were isolated and cultured for a total of 14 days in serum containing medium. The organization of calcium-handling proteins and myofibrillar structure in freshly isolated and in long-term cultured adult cardiomyocytes was studied by immunofluorescence and electron microscopy. Excitation contraction-coupling was analyzed by fura 2 and video edge detection in electrically paced cardiomyocytes forming a monolayer, and cell death and viability was measured by TUNEL assay, LDH release, MTT assay, and Western blot for LC3. Adult cardiomyocytes treated with Doxo showed apoptosis and necrosis only at supraclinical concentrations. Treated cells displayed merely alterations in cytoskeleton organization and integrity concomitant with contractile dysfunction and up-regulation of autophagosome formation, but no change in total sarcomeric protein content. We propose, that myofibrillar damage contributes to contractile dysfunction prior to cell death in adult cardiomyocytes exposed to clinically relevant concentrations of anthracyclines.

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Ultrastructural morphometric analysis of cultured neonatal and adult rat ventricular cardiac muscle cells

1989

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Neonatal and adult rat ventricular cardiac muscle cells cultured on laminin differed from similar myocytes grown on plastic in the amount and distribution of their mitochondria and transverse tubules. Point-count morphometry was used at the electron microscopic level to quantify these differences. Adult myocytes grown on laminin contained more mitochondria per unit volume than adult myocytes grown on plastic. No significant differences were observed in the volume percent of myofibrils in either adult or neonatal ventricular myocytes when grown on laminin and compared to those grown on plastic. The transverse tubule system in neonatal and adult myocytes was reduced significantly when both groups were cultured on laminin. Furthermore, neonatal and adult myocytes cultured on laminin were flatter than those cultured on plastic. This may indicate a relationship between the surface/volume ratio and transverse tubule development in cultured myocytes. These studies establish that point-count morphometry can be used to quantify changes in the organelle volume densities of cultured cardiac muscle cells.

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Immunocytochemical analysis of the regeneration of myofibrils in long-term cultures of adult cardiomyocytes of the rat

Cited by 87 publications

References 56 publications

Intracompartmental sorting of essential myosin light chains: Molecular dissection and in vivo monitoring by epitope tagging

Intracompartmental sorting of essential myosin light chains: Molecular dissection and in vivo monitoring by epitope tagging

The role of cell death and myofibrillar damage in contractile dysfunction of long-term cultured adult cardiomyocytes exposed to doxorubicin

Ultrastructural morphometric analysis of cultured neonatal and adult rat ventricular cardiac muscle cells

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