Myoblast fusion requires fibronectin degradation by exteriorized m-calpain

N Dourdin, J J Brustis, Denis Pierre Balcerzak, N Elamrani, S Poussard, P Cottin, A Ducastaing

    Research output: Contribution to journalArticle

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    Abstract

    We recently reported that when myoblasts fuse, m-calpain could be exteriorized. Indeed, at present a number of works support this hypothesis because this enzyme was localized intercellularly and more particularly associated to extracellular matrix components. Knowing that the cell surface of the fusing myoblast is supposed to undergo many changes, we addressed the question whether m-calpain could be involved in the phenomenon of fusion via fibronectin cleavage or degradation. Using different digestion experiments, we demonstrated that soluble purified fibronectin and highly insoluble fibronectin fibrils represent very good substrates for this proteinase; moreover, at the burst of fusion, fibronectin proteolytic fragments could be identified. On the other hand, we have conducted biological assays on cultured myoblasts using a defined medium supplemented by exogenous factors capable of stimulating or inhibiting m-calpain activity. The effects of such factors on rat myoblast fusion and concomitantly on the targeted glycoprotein were analyzed and quantified. When m-calpain activity and the phenomenon of fusion were reduced (defined medium without insulin), the amount of the 220-kDa fibronectin band was increased by 43%. When m-calpain activity and myoblast fusion were prevented by addition of antibodies to m-calpain or calpain inhibitor II, the fibronectin concentration was higher since it was increased by similar to 67 and similar to 71%, respectively. In addition, when observed at the ultrastructural level, m-calpain seems to be localized at the potential fusion site of myoblasts and more particularly associated to the extracellular matrix when muscle cells were initially treated by anti-m-calpain IgG. Taken together, these results support the hypothesis that exteriorized m-calpain could be, in part, involved in myoblast fusion via fibronectin alteration or degradation. (C) 1997 Academic Press.

    Original languageEnglish
    Pages (from-to)385-394
    Number of pages10
    JournalExperimental Cell Research
    Volume235
    Issue number2
    Publication statusPublished - 15 Sep 1997

    Keywords

    • CYSTEINE PROTEINASE CALPAIN
    • CALCIUM-ACTIVATED PROTEASE
    • ARTHRITIC SYNOVIAL JOINTS
    • RABBIT SKELETAL-MUSCLE
    • MYOGENIC DIFFERENTIATION
    • BIOCHEMICAL DEMONSTRATION
    • EXTRACELLULAR-MATRIX
    • CELLS
    • CLEAVAGE
    • SURFACE

    Cite this

    Dourdin, N., Brustis, J. J., Balcerzak, D. P., Elamrani, N., Poussard, S., Cottin, P., & Ducastaing, A. (1997). Myoblast fusion requires fibronectin degradation by exteriorized m-calpain. Experimental Cell Research, 235(2), 385-394.

    Myoblast fusion requires fibronectin degradation by exteriorized m-calpain. / Dourdin, N ; Brustis, J J ; Balcerzak, Denis Pierre; Elamrani, N ; Poussard, S ; Cottin, P ; Ducastaing, A .

    In: Experimental Cell Research, Vol. 235, No. 2, 15.09.1997, p. 385-394.

    Research output: Contribution to journalArticle

    Dourdin, N, Brustis, JJ, Balcerzak, DP, Elamrani, N, Poussard, S, Cottin, P & Ducastaing, A 1997, 'Myoblast fusion requires fibronectin degradation by exteriorized m-calpain' Experimental Cell Research, vol. 235, no. 2, pp. 385-394.
    Dourdin N, Brustis JJ, Balcerzak DP, Elamrani N, Poussard S, Cottin P et al. Myoblast fusion requires fibronectin degradation by exteriorized m-calpain. Experimental Cell Research. 1997 Sep 15;235(2):385-394.
    Dourdin, N ; Brustis, J J ; Balcerzak, Denis Pierre ; Elamrani, N ; Poussard, S ; Cottin, P ; Ducastaing, A . / Myoblast fusion requires fibronectin degradation by exteriorized m-calpain. In: Experimental Cell Research. 1997 ; Vol. 235, No. 2. pp. 385-394.
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    abstract = "We recently reported that when myoblasts fuse, m-calpain could be exteriorized. Indeed, at present a number of works support this hypothesis because this enzyme was localized intercellularly and more particularly associated to extracellular matrix components. Knowing that the cell surface of the fusing myoblast is supposed to undergo many changes, we addressed the question whether m-calpain could be involved in the phenomenon of fusion via fibronectin cleavage or degradation. Using different digestion experiments, we demonstrated that soluble purified fibronectin and highly insoluble fibronectin fibrils represent very good substrates for this proteinase; moreover, at the burst of fusion, fibronectin proteolytic fragments could be identified. On the other hand, we have conducted biological assays on cultured myoblasts using a defined medium supplemented by exogenous factors capable of stimulating or inhibiting m-calpain activity. The effects of such factors on rat myoblast fusion and concomitantly on the targeted glycoprotein were analyzed and quantified. When m-calpain activity and the phenomenon of fusion were reduced (defined medium without insulin), the amount of the 220-kDa fibronectin band was increased by 43{\%}. When m-calpain activity and myoblast fusion were prevented by addition of antibodies to m-calpain or calpain inhibitor II, the fibronectin concentration was higher since it was increased by similar to 67 and similar to 71{\%}, respectively. In addition, when observed at the ultrastructural level, m-calpain seems to be localized at the potential fusion site of myoblasts and more particularly associated to the extracellular matrix when muscle cells were initially treated by anti-m-calpain IgG. Taken together, these results support the hypothesis that exteriorized m-calpain could be, in part, involved in myoblast fusion via fibronectin alteration or degradation. (C) 1997 Academic Press.",
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    AU - Dourdin, N

    AU - Brustis, J J

    AU - Balcerzak, Denis Pierre

    AU - Elamrani, N

    AU - Poussard, S

    AU - Cottin, P

    AU - Ducastaing, A

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    N2 - We recently reported that when myoblasts fuse, m-calpain could be exteriorized. Indeed, at present a number of works support this hypothesis because this enzyme was localized intercellularly and more particularly associated to extracellular matrix components. Knowing that the cell surface of the fusing myoblast is supposed to undergo many changes, we addressed the question whether m-calpain could be involved in the phenomenon of fusion via fibronectin cleavage or degradation. Using different digestion experiments, we demonstrated that soluble purified fibronectin and highly insoluble fibronectin fibrils represent very good substrates for this proteinase; moreover, at the burst of fusion, fibronectin proteolytic fragments could be identified. On the other hand, we have conducted biological assays on cultured myoblasts using a defined medium supplemented by exogenous factors capable of stimulating or inhibiting m-calpain activity. The effects of such factors on rat myoblast fusion and concomitantly on the targeted glycoprotein were analyzed and quantified. When m-calpain activity and the phenomenon of fusion were reduced (defined medium without insulin), the amount of the 220-kDa fibronectin band was increased by 43%. When m-calpain activity and myoblast fusion were prevented by addition of antibodies to m-calpain or calpain inhibitor II, the fibronectin concentration was higher since it was increased by similar to 67 and similar to 71%, respectively. In addition, when observed at the ultrastructural level, m-calpain seems to be localized at the potential fusion site of myoblasts and more particularly associated to the extracellular matrix when muscle cells were initially treated by anti-m-calpain IgG. Taken together, these results support the hypothesis that exteriorized m-calpain could be, in part, involved in myoblast fusion via fibronectin alteration or degradation. (C) 1997 Academic Press.

    AB - We recently reported that when myoblasts fuse, m-calpain could be exteriorized. Indeed, at present a number of works support this hypothesis because this enzyme was localized intercellularly and more particularly associated to extracellular matrix components. Knowing that the cell surface of the fusing myoblast is supposed to undergo many changes, we addressed the question whether m-calpain could be involved in the phenomenon of fusion via fibronectin cleavage or degradation. Using different digestion experiments, we demonstrated that soluble purified fibronectin and highly insoluble fibronectin fibrils represent very good substrates for this proteinase; moreover, at the burst of fusion, fibronectin proteolytic fragments could be identified. On the other hand, we have conducted biological assays on cultured myoblasts using a defined medium supplemented by exogenous factors capable of stimulating or inhibiting m-calpain activity. The effects of such factors on rat myoblast fusion and concomitantly on the targeted glycoprotein were analyzed and quantified. When m-calpain activity and the phenomenon of fusion were reduced (defined medium without insulin), the amount of the 220-kDa fibronectin band was increased by 43%. When m-calpain activity and myoblast fusion were prevented by addition of antibodies to m-calpain or calpain inhibitor II, the fibronectin concentration was higher since it was increased by similar to 67 and similar to 71%, respectively. In addition, when observed at the ultrastructural level, m-calpain seems to be localized at the potential fusion site of myoblasts and more particularly associated to the extracellular matrix when muscle cells were initially treated by anti-m-calpain IgG. Taken together, these results support the hypothesis that exteriorized m-calpain could be, in part, involved in myoblast fusion via fibronectin alteration or degradation. (C) 1997 Academic Press.

    KW - CYSTEINE PROTEINASE CALPAIN

    KW - CALCIUM-ACTIVATED PROTEASE

    KW - ARTHRITIC SYNOVIAL JOINTS

    KW - RABBIT SKELETAL-MUSCLE

    KW - MYOGENIC DIFFERENTIATION

    KW - BIOCHEMICAL DEMONSTRATION

    KW - EXTRACELLULAR-MATRIX

    KW - CELLS

    KW - CLEAVAGE

    KW - SURFACE

    M3 - Article

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    SP - 385

    EP - 394

    JO - Experimental Cell Research

    JF - Experimental Cell Research

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    ER -