Myofibrillogenesis and formation of cell contacts mediate the localization of N-RAP in cultured chick cardiomyocytes

Carroll, Stefanie; Horowits, Robert

doi:10.1002/1097-0169(200009)47:1<63::aid-cm6>3.0.co;2-m

Cited by 30 publications

(41 citation statements)

References 23 publications

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“…Mouse embryonic cardiomyocytes were cultured as described previously with minor modifications [22,26]. Briefly, embryonic day 17.5 hearts were isolated and cultured by successive digestion with 0.25% trypsin (Gibco) and 0.2% collagenase type II (Sigma).…”

Section: Primary Culture and Transfection Of Embryonic Mouse Cardiomymentioning

confidence: 99%

“…Our previous studies on cardiomyocytes isolated from chick or mouse hearts at embryonic day 17 demonstrated that preexisting myofibrils are largely disassembled when the cells are dissociated and plated, and that new myofibrils assemble as the cardiomyocytes spread in culture [22,25,26]. However, on subsequent days the cultured cardiomyocytes are heterogeneous with respect to cell size, shape, and myofibril organization.…”

Section: Krp1 Promotes Lateral Fusion Of Myofibril Assembly Intermedimentioning

confidence: 99%

“…These include N-RAP, a LIM domain and nebulin repeat protein found in the premyofibril regions of cardiomyocytes and at the Zlines and M-lines of newly formed myofibrils [14,22,23], and Krp1, which binds N-RAP and is found between laterally fusing myofibrils in spreading chick cardiomyocytes [14]. During myofibril assembly, N-RAP organizes actin and α-actinin into I-Z-I structures (symmetrical actin filaments with their barbed ends anchored at a central Z-body or Z-line containing α-actinin) [24,25], while Krp1 has been hypothesized to promote lateral fusion of premyofibril structures [14].…”

Section: Introductionmentioning

confidence: 99%

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Krp1 (Sarcosin) promotes lateral fusion of myofibril assembly intermediates in cultured mouse cardiomyocytes

Greenberg

Connelly

Daniels

et al. 2008

Experimental Cell Research

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Krp1, also called sarcosin, is a cardiac and skeletal muscle kelch-repeat protein hypothesized to promote the assembly of myofibrils, the contractile organelles of striated muscles, through interaction with N-RAP and actin. To elucidate its role, endogenous Krp1 was studied in primary embryonic mouse cardiomyocytes. While immunofluorescence showed punctate Krp1 distribution throughout the cell, detergent extraction revealed a significant pool of Krp1 associated with cytoskeletal elements. Reduction of Krp1 expression with siRNA resulted in specific inhibition of myofibril accumulation with no effect on cell spreading. Immunostaining analysis and electron microscopy revealed that cardiomyocytes lacking Krp1 contained sarcomeric proteins with longitudinal periodicities similar to mature myofibrils, but fibrils remained thin and separated. These thin myofibrils were degraded by a scission mechanism distinct from the myofibril disassembly pathway observed during cell division in the developing heart. The data are consistent with a model in which Krp1 promotes lateral fusion of adjacent thin fibrils into mature, wide myofibrils and contribute insight into mechanisms of myofibrillogenesis and disassembly.

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Section: Primary Culture and Transfection Of Embryonic Mouse Cardiomymentioning

confidence: 99%

Section: Krp1 Promotes Lateral Fusion Of Myofibril Assembly Intermedimentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Krp1 (Sarcosin) promotes lateral fusion of myofibril assembly intermediates in cultured mouse cardiomyocytes

Greenberg

Connelly

Daniels

et al. 2008

Experimental Cell Research

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“…Although subcellular localization and biochemical copurification from adult muscles suggested that N-RAP may link the ends of myofibrils to membrane-associated protein complexes, studies in cultured embryonic cardiomyocytes have implicated N-RAP in myofibril assembly (Carroll and Horowits, 2000;Carroll et al, 2001Carroll et al, , 2004Lu et al, 2003). These studies showed that N-RAP localizes with the earliest myofibril precursors that originate near the membrane (Carroll and Horowits, 2000;Lu et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

“…These studies showed that N-RAP localizes with the earliest myofibril precursors that originate near the membrane (Carroll and Horowits, 2000;Lu et al, 2003). These precursors contain punctate ␣-actinin Z-bodies, ␣-actin, and muscle tropomyosin (Dlugosz et al, 1984;Wang et al, 1988;Schultheiss et al, 1990;Handel et al, 1991;Rhee et al, 1994;Dabiri et al, 1997), as well as nonmuscle myosin IIb (Rhee et al, 1994).…”

Section: Introductionmentioning

confidence: 99%

N-RAP expression during mouse heart development

Borst

Horowits

2005

Dev. Dyn.

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N-RAP gene expression and N-RAP localization were studied during mouse heart development using semiquantitative reverse transcriptase-polymerase chain reaction and immunofluorescence. N-RAP mRNA was detected at embryonic day (E) 10.5, significantly increased from E10.5 to E16.5, and remained essentially constant from E16.5 until 21 days after birth. In E9.5-10.5 heart tissue, N-RAP protein was primarily associated with developing premyofibril structures containing ␣-actinin,

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ALP and MLP distribution during myofibrillogenesis in cultured cardiomyocytes

Henderson

Pomiès

Auffray

et al. 2003

Cell Motil. Cytoskeleton

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The Z-line is a multifunctional macromolecular complex that anchors sarcomeric actin filaments, mediates interactions with intermediate filaments and costameres, and recruits signaling molecules. Antiparallel alpha-actinin homodimers, present at Z-lines, cross-link overlapping actin filaments and also bind other cytoskeletal and signaling elements. Two LIM domain containing proteins, alpha-actinin associated LIM protein (ALP) and muscle LIM protein (MLP), interact with alpha-actinin, distribute in vivo to Z-lines or costameres, respectively, and, when absent, are associated with heart disease. Here we describe the behavior of ALP and MLP during myofibrillogenesis in cultured embryonic chick cardiomyocytes. As myofibrils develop, ALP and MLP are observed in distinct distribution patterns in the cell. ALP is coincident with alpha-actinin from the first stage of myofibrillogenesis and co-distributes with alpha-actinin to Z-lines and intercalated discs in mature myofibrils. Interestingly, we also demonstrate using ALP-GFP transfection experiments and an in vitro binding assay that the ALP-alpha-actinin binding interaction is not required to target ALP to the Z-line. In contrast, MLP localization is not co-incident with that of alpha-actinin until late stages of myofibrillogenesis; however, it is present in premyofibrils and nascent myofibrils prior to the incorporation of other costameric components such as vinculin, vimentin, or desmin. Our observations support the view that ALP function is required specifically at actin anchorage sites. The subcellular distribution pattern of MLP during myofibrillogenesis suggests that it functions during differentiation prior to the establishment of costameres.

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Myofibrillogenesis and formation of cell contacts mediate the localization of N-RAP in cultured chick cardiomyocytes

Cited by 30 publications

References 23 publications

Krp1 (Sarcosin) promotes lateral fusion of myofibril assembly intermediates in cultured mouse cardiomyocytes

Krp1 (Sarcosin) promotes lateral fusion of myofibril assembly intermediates in cultured mouse cardiomyocytes

N-RAP expression during mouse heart development

ALP and MLP distribution during myofibrillogenesis in cultured cardiomyocytes

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