Three‐dimensional analysis and visualization of myofibrillogenesis in adult cardiomyocytes by confocal microscopy

Messerli, J. M.; Perriard, Jean‐Claude

doi:10.1002/jemt.1070300609

Cited by 17 publications

(7 citation statements)

References 67 publications

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“…ARC in primary culture reexpress the pacemaker current I f (27), which presumably drives contractile activity after day 7 and consequently may contribute to restoration of Cbl signaling. Another possible, though speculative, explanation for inefficiency of the Akt pathway to induce translocation of GLUT4 is disarray of the actin-based cytoskeletal structure, which occurs in cultured ARC from day 2 to 4 (28). In adipocytes, GLUT4 translocation is dependent on actincontaining cytoskeletal components (29).…”

Section: Translocation Of Glut4 and Signaling Cascadesmentioning

confidence: 99%

Insulin resistance in adult cardiomyocytes undergoing dedifferentiation: role of GLUT4 expression and translocation

et al. 2004

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Myocardium undergoing remodeling in vivo exhibits insulin resistance that has been attributed to a shift from the insulin-sensitive glucose transporter GLUT4 to the fetal, less insulin-sensitive, isoform GLUT1. To elucidate the role of altered GLUT4 expression in myocardial insulin resistance, glucose uptake and the expression of the glucose transporter isoforms GLUT4 and GLUT1 were measured in adult rat cardiomyocytes (ARC). ARC in culture spontaneously undergo dedifferentiation, hypertrophy-like spreading, and return to a fetal-like gene expression pattern. Insulin stimulation of 2-deoxy-D-glucose uptake was completely abolished on day 2 and 3 of culture and recovered thereafter. Although GLUT4 protein level was reduced, the time-course of unresponsiveness to insulin did not correlate with altered expression of GLUT1 and GLUT4. However, translocation of GLUT4 to the sarcolemma in response to insulin was completely abolished during transient insulin resistance. Insulin-mediated phosphorylation of Akt was not reduced, indicating that activation of phosphatidylinositol 3-kinase (PI3K) was preserved. On the other hand, total and phosphorylated Cbl was reduced during insulin resistance, suggesting that activation of Cbl/CAP is essential for insulin-mediated GLUT4 translocation, in addition to activation of PI3K. Pharmacological inhibition of contraction in insulin-sensitive ARC reduced insulin sensitivity and lowered phosphorylated Cbl. The results suggest that transient insulin resistance in ARC is related to impairment of GLUT4 translocation. A defect in the PI3K-independent insulin signaling pathway involving Cbl seems to contribute to reduced insulin responsiveness and may be related to contractile arrest.

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Section: Translocation Of Glut4 and Signaling Cascadesmentioning

confidence: 99%

Insulin resistance in adult cardiomyocytes undergoing dedifferentiation: role of GLUT4 expression and translocation

et al. 2004

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“…The study of many transparent biological specimens and their organization is an intrinsically three-dimensional (3D) endeavor (Charrière et al, 2006a;Fauver et al, 2005;Ferko et al, 2006;Heise et al, 2005;Messerli and Perriard, 1995). In particular, the observation of transparent phase objects and the 3D refractive index (RI) distribution are subjects of long-standing interests in the imaging community.…”

Section: Introductionmentioning

confidence: 99%

Three‐dimensional refractive index reconstruction with quantitative phase tomography

Dragomir

Goh

Roberts

2007

Microscopy Res & Technique

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Optical tomography based on quantitative phase microscopy is used to determine nondestructively and with high spatial resolution the three-dimensional (3D) refractive index distributions within optical fiber devices. After obtaining a series of phase images of the fiber as it is rotated around its longitudinal axis at regularly-spaced angular positions, filtered backprojection is used to reconstruct a 3D map of the refractive index. The 3D refractive index distribution of the join region between two fusion spliced optical fibers is reconstructed with accuracy better than 10(-3).

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“…Technical developments of fluorescent probes including green-fluorescent protein [1]and its variants [2], and increased scanning speed of microscope systems have added significantly to studies of intracellular dynamics in living cells. So far, spatial distribution of cellular components [3], membrane dynamics [4], fusion [5]and transport of vesicles [6]have been visualized with time-lapse recording. Furthermore, fluorescence-tagged single particles can now be detected in time-lapse studies [7].…”

Section: Introductionmentioning

confidence: 99%

In vivo Confocal Laser Scanning Microscopy and Micropuncture in Intact Rat

Ohno¹,

Birn²,

Christensen³

2005

Nephron Exp Nephrol

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Background: Intravital microscopy theoretically provides the optimal conditions for studying specific organ functions. However, the application of microscopy in intact organs in vivo has been limited so far due to technical difficulties. The purpose of this study was to establish a method of in vivo confocal laser scanning microscopy (CLSM) for the study of endocytosis in proximal tubules of intact kidney. Methods: The left kidney of rats placed on a modified microscope stage was exposed and stabilized in a thermostatically controlled cup. The stage was then attached to an upright confocal microscope. Surface proximal tubules were microinfused with fluorescent albumin or transferrin. Single or time-series images of microinfused proximal tubules were recorded in reflection and/or fluorescence mode. Results: The stability of the kidney and the resolution of images were sufficient to visualize intracellular vesicles. Albumin and transferrin were initially observed at the brush border, then later internalized by proximal tubules and accumulated in lysosomes over a time period of 15 min. Furthermore, fusion of vesicles was observed in time-lapse images. Conclusion: These results show that in vivo CLSM in intact kidney may be an excellent method to evaluate proximal tubular endocytosis and ligand trafficking.

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Three‐dimensional analysis and visualization of myofibrillogenesis in adult cardiomyocytes by confocal microscopy

Cited by 17 publications

References 67 publications

Insulin resistance in adult cardiomyocytes undergoing dedifferentiation: role of GLUT4 expression and translocation

Insulin resistance in adult cardiomyocytes undergoing dedifferentiation: role of GLUT4 expression and translocation

Three‐dimensional refractive index reconstruction with quantitative phase tomography

In vivo Confocal Laser Scanning Microscopy and Micropuncture in Intact Rat

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