Phospholipase D is involved in myogenic differentiation through remodeling of actin cytoskeleton.
We investigated the role of phospholipase D (PLD) and its product phosphatidic acid (PA) in myogenic differentiation of cultured L6 rat skeletal myoblasts. Arginine-vasopressin (AVP), a differentiation inducer, rapidly activated PLD in a Rho-dependent way, as shown by almost total suppression of activation by C3 exotoxin pretreatment. Addition of 1-butanol, which selectively inhibits PA production by PLD, markedly decreased AVP-induced myogenesis. Conversely, myogenesis was potentiated by PLD1b isoform overexpression but not by PLD2 overexpression, establishing that PLD1 is involved in this process. The expression of the PLD isoforms was differentially regulated during differentiation. AVP stimulation of myoblasts induced the rapid formation of stress fiber-like actin structures (SFLSs). 1-Butanol selectively inhibited this response, whereas PLD1b overexpression induced SFLS formation, showing that it was PLD dependent. Endogenous PLD1 was located at the level of SFLSs, and by means of an intracellularly expressed fluorescent probe, PA was shown to be accumulated along these structures in response to AVP. In addition, AVP induced a PLD-dependent neosynthesis of phosphatidylinositol 4,5-bisphosphate (PIP2), which also was accumulated along actin fibers. These data support the hypothesis that PLD participates in myogenesis through PA- and PIP2-dependent actin fiber formation.
Komati
Hiba
H
Laboratoire de Physiopathologie des Lipides et Membranes, Institut National de la Santé et de la Recherche Médicale Unité 585, Institut National des Sciences Appliquées de Lyon, 69621 Villeurbanne, France.
Naro
Fabio
F
Mebarek
Saida
S
De Arcangelis
Vania
V
Adamo
Sergio
S
Lagarde
Michel
M
Prigent
Annie-France
AF
Némoz
Georges
G
D020001:1-Butanol / Q000737:chemistry; D000199:Actins / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D001127:Arginine Vasopressin / Q000737:chemistry; D002454:Cell Differentiation; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism*; D015536:Down-Regulation; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D018482:Muscle, Skeletal / Q000166:cytology; D009132:Muscles / Q000166:cytology*; D018008:Myogenin / Q000378:metabolism; D010590:Phalloidine / Q000494:pharmacology; D010641:Phenotype; D010712:Phosphatidic Acids / Q000737:chemistry; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000737:chemistry; D010739:Phospholipase D / Q000378:metabolism / Q000502:physiology*; D010957:Plasmids / Q000378:metabolism; D020033:Protein Isoforms; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014162:Transfection; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
D000199:Actins; C503803:Myog protein, rat; D018008:Myogenin; D010712:Phosphatidic Acids; D019269:Phosphatidylinositol 4,5-Diphosphate; D020033:Protein Isoforms; D001127:Arginine Vasopressin; D010590:Phalloidine; D020001:1-Butanol; C105606:phospholipase D2; D010739:Phospholipase D; C106801:phospholipase D1; D020742:rhoA GTP-Binding Protein