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Muscle and neuronal differentiation in primary cell culture of larval Mytilus trossulus (Mollusca: Bivalvia)

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Abstract

Molluscan in vitro technology allows the study of the differentiation of isolated cells undergoing experimental manipulations. We have used the immunofluorescence technique and laser scanning microscopy to investigate the organization of muscle proteins (actin, myosin, paramyosin, and twitchin) and the localization of neurotransmitters (serotonin and FMRFamide) in cultured mussel larval cells. Differentiation into muscle and neuron-like cells occurs during the cultivation of mussel cells from premyogenic and prenervous larval stages. Muscle proteins are colocalized in contractile cells through all stages of cultivation. The cultivation of mussel cells on various substrates and the application of integrin receptor blockers suggest that an integrin-dependent mechanism is involved in cell adhesion and differentiation. Dissociated mussel cells aggregate and become self-organized in culture. After 20 days of cultivation, they form colonies in which serotonin- and FMRFamide-immunoreactive cells are located centrally, whereas muscle cells form a contractile network at the periphery. The pattern of thick and thin filaments in cultivated mussel cells changes according to the scenario of muscle arrangement in vivo: initially, a striated pattern of muscle filaments forms but is then replaced by a smooth muscle pattern with a diffuse distribution of muscle proteins, typical of muscles of adult molluscs. Myogenesis in molluscs thus seems to be a highly dynamic and potentially variable process. Such a “flexible” developmental program can be regarded as a prerequisite for the evolution of the wide variety of striated and smooth muscles in larval and adult molluscs.

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Acknowledgements

The authors are grateful to Dr. Elena Voronezhskaya (Institute of Developmental Biology, Russian Academy of Sciences, Moscow, Russia) for help with the immunocytochemistry and for valuable discussions and criticisms during the preparation of the manuscript and to Mrs. Irina Barsegova for her help in editing the manuscript. The donation of thick filament proteins by Dr. N.S. Shelud’ko and Dr. O.S. Matusovsky (Zhirmunsky Institute of Marine Biology, Far Eastern Branch of the Russian Academy of Sciences, Vladivostok) is gratefully acknowledged. The authors are particularly grateful to the valuable comments of anonymous reviewers who helped to improve the manuscript.

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Authors and Affiliations

  1. Laboratory of Cytotechnology, A.V. Zhirmunsky Institute of Marine Biology, Far Eastern Branch of the Russian Academy of Sciences, Palchevsky Street 17, 690041, Vladivostok, Russia

    Nelly A. Odintsova & Vyacheslav A. Dyachuk

  2. Laboratory of Comparative Physiology, Institute of Developmental Biology, Russian Academy of Sciences, Vavilov Street 26, 119991, Moscow, Russia

    Leonid P. Nezlin

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  1. Nelly A. Odintsova
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  2. Vyacheslav A. Dyachuk
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  3. Leonid P. Nezlin
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Correspondence to Nelly A. Odintsova.

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This study was supported in part by the Far Eastern Branch of Russian Academy of Sciences (grants NT-08-016-04, 09-III-B-06-252, 09-II-SB-06-001, 09-I-P22-04), RFBR (grants 09-04-98529-r_vostok_а and 09-04-01326-а) and President Grant to V.A. Dyachuk (MK-2425.2010.4).

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Odintsova, N.A., Dyachuk, V.A. & Nezlin, L.P. Muscle and neuronal differentiation in primary cell culture of larval Mytilus trossulus (Mollusca: Bivalvia). Cell Tissue Res 339, 625–637 (2010). https://doi.org/10.1007/s00441-009-0918-3

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