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1.
Cytokine ; 55(2): 280-7, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21570315

RESUMO

Muscle fibers are formed during embryonic development by the fusion of mononucleated myoblasts. The spatial structure and molecular composition of the sarcolemma are crucial for the myoblast recognition and fusion steps. Cyclodextrins are a group of substances that have the ability to solubilize lipids through the formation of molecular inclusion complexes. Previously, we have shown that methyl-ß-cyclodextrin (MbCD) enhances muscle differentiation. Here, we analyzed the effects of α-cyclodextrin (aCD) during myogenesis. Myogenic cultures treated with aCD showed an increase in myoblast fusion and in the expression of myogenin, sarcomeric tropomyosin and desmin. aCD-conditioned media accelerates myogenesis in a similar way as aCD does, and increased levels of IL-4 were found in aCD-conditioned media. aCD-induced effects on myogenesis were inhibited by an anti-IL4 antibody. These results show that α-cyclodextrin induces myogenic differentiation by the release of IL-4.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Fusão Celular , Interleucina-4/metabolismo , Mioblastos/efeitos dos fármacos , Mioblastos/metabolismo , Mioblastos/fisiologia , alfa-Ciclodextrinas/farmacologia , Animais , Células Cultivadas , Embrião de Galinha , Meios de Cultivo Condicionados/química , Meios de Cultivo Condicionados/metabolismo , Desenvolvimento Muscular/efeitos dos fármacos , Músculo Esquelético/citologia , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/embriologia , Mioblastos/citologia
2.
Cells Tissues Organs ; 192(3): 187-99, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20523034

RESUMO

Cholesterol is a sterol lipid that plays pleiotropic roles in the plasma membrane; it is involved in maintaining membrane fluidity and permeability and the structure of lipid microdomains. Despite its importance, the consequences of membrane cholesterol depletion during cardiac differentiation have not been described. Therefore, we investigated the cellular and molecular mechanisms associated with cholesterol depletion in cultures of chick cardiac cells. We used methyl-beta-cyclodextrin (MCD) to deplete membrane cholesterol and investigate its role in cardiac differentiation by following the expression of several markers including the transcriptional factor Nkx2.5, the myofibrillar protein tropomyosin, the cytoskeletal intermediate filament protein desmin, the caveolar protein caveolin-3, the cadherin/beta-catenin adhesion complex, and the junctional protein connexin 43. Confocal microscopy showed that desmin-positive cells were located more externally in the aggregates in relation to the more internally located caveolin-3-positive cells. Desmin and caveolin-3 were co-localized in filamentous structures in the subsarcolemmal region of well-spread cells outside the aggregates. beta-Catenin was concentrated in regions of cell-cell contact, and tropomyosin in sarcomeric structures. Western blot tests showed that immediately following cholesterol depletion, there was a slight decrease in the expression of caveolin-3 and desmin, and at the same time there was a sharp increase in the expression of cadherin, tropomyosin, Nkx2.5 and connexin 43. Further, we found an increase in the expression of cardiac beta-myosin heavy chain 7, a marker of the cardiac hypertrophic phenotype. These observations suggest that membrane cholesterol plays a significant role in regulating cardiomyocyte differentiation.


Assuntos
Antígenos de Diferenciação/metabolismo , Diferenciação Celular/fisiologia , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Colesterol/metabolismo , Miócitos Cardíacos/metabolismo , beta-Ciclodextrinas/farmacologia , Animais , Caderinas/metabolismo , Miosinas Cardíacas/metabolismo , Caveolina 3/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Embrião de Galinha , Conexina 43/metabolismo , Meios de Cultivo Condicionados/metabolismo , Citoplasma/metabolismo , Desmina/metabolismo , Miócitos Cardíacos/citologia , Miócitos Cardíacos/efeitos dos fármacos , Cadeias Pesadas de Miosina/metabolismo , Sarcômeros/metabolismo , Fatores de Transcrição/metabolismo , Tropomiosina/metabolismo , beta Catenina/metabolismo
3.
PLoS One ; 7(5): e38147, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22662278

RESUMO

Cardiac cells are organized in vivo in a complex tridimensional structural organization that is crucial for heart function. While in vitro studies can reveal details about cardiac cell biology, usually cells are grown on simplified two-dimensional (2D) environments. To address these differences, we established a cardiac cell culture composed of both 2D and three-dimensional (3D)-organized cells. Our results shows significant differences between the two culture contexts in relation to the overall morphology of the cells, contraction ability, proliferation rate, presence of intercellular adhesion structures, organization of myofibrils, mitochondria morphology, endoplasmic reticulum contents, cytoskeletal filaments and extracellular matrix distribution, and expression of markers of cardiac differentiation. Cardiac cells grown in 2D-context displayed a flattened and well spread shape, were mostly isolated and their cytoplasm was filled with a large network of microfilaments and microtubules. In contrast, 3D-cells were smaller in size, were always in close contact with each other with several cellular junctions, and displayed a less conspicuous cytoskeletal network. 3D-cells had more mitochondria and myofibrils and these cells contract spontaneously more often than 2D-cells. On the other hand, endoplasmic reticulum membranes were present in higher amounts in 2D-cells when compared to 3D-cells. The expression of desmin, cadherin and alpha-actinin was higher in 3D-aggregates compared to 2D-spread cells. These findings indicate that the tridimensional environment in which the cardiac cells are grown influence several aspects of cardiac differentiation, including cell adhesion, cell shape, myofibril assembly, mitochondria contents and protein expression. We suggest that the use of this cardiac culture model, with 2D and 3D-context cells, could be useful for studies on the effects of different drugs, or growth factors, giving valuable information on the biological response of cells grown in different spatial organizations.


Assuntos
Junções Aderentes/metabolismo , Miócitos Cardíacos/citologia , Miócitos Cardíacos/metabolismo , Cultura Primária de Células/métodos , Actinina/metabolismo , Animais , Caderinas/metabolismo , Proliferação de Células , Embrião de Galinha , Citoesqueleto/metabolismo , Desmina/metabolismo , Microtúbulos/metabolismo , Contração Miocárdica , Miócitos Cardíacos/ultraestrutura , Miofibrilas/metabolismo , Miofibrilas/ultraestrutura
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