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Differential expression and localization of Ankrd2 isoforms in human skeletal and cardiac muscles

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Abstract

Four human Ankrd2 transcripts, reported in the Ensembl database, code for distinct protein isoforms (360, 333, 327 and 300 aa), and so far, their existence, specific expression and localization patterns have not been studied in detail. Ankrd2 is preferentially expressed in the slow fibers of skeletal muscle. It is found in both the nuclei and the cytoplasm of skeletal muscle cells, and its localization is prone to change during differentiation and upon stress. Ankrd2 has also been detected in the heart, in ventricular cardiomyocytes and in the intercalated disks (ICDs). The main objective of this study was to distinguish between the Ankrd2 isoforms and to determine the contribution of each one to the general profile of Ankrd2 expression in striated muscles. We demonstrated that the known expression and localization pattern of Ankrd2 in striated muscle can be attributed to the isoform of 333 aa which is dominant in both tissues, while the designated cardiac and canonical isoform of 360 aa was less expressed in both tissues. The 360 aa isoform has a distinct nuclear localization in human skeletal muscle, as well as in primary myoblasts and myotubes. In contrast to the isoform of 333 aa, it was not preferentially expressed in slow fibers and not localized to the ICDs of human cardiomyocytes. Regulation of the expression of both isoforms is achieved at the transcriptional level. Our results set the stage for investigation of the specific functions and interactions of the Ankrd2 isoforms in healthy and diseased human striated muscles.

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Acknowledgments

This study was supported by a grant from the Ministry of Education, Science and Technological Development of the Republic of Serbia (Project No. 173008 to DR), a bilateral project funded by the German Academic Exchange Service and the Ministry of Education, Science and Technological Development of the Republic of Serbia (Project-ID 50753561 – ANDIMA to SK and MCW) and a grant from the Fondazione Cariparo, Italy (Progetto Eccellenza 2010 CHROMUS to GV and GF). We thank the Muscle Tissue Culture Collection (MTCC) for providing the myoblast samples. The Muscle Tissue Culture Collection is part of the German network on muscular dystrophies (MD-NET) and the German network for mitochondrial disorders (mito-NET, project D2, 01GM1113A) funded by the German Ministry of Education and Research (BMBF, Bonn, Germany). The Muscle Tissue Culture Collection is a partner of the EuroBioBank (www.eurobiobank.org) and the TREAT-NMD (www.treat-nmd.eu). We are grateful to Natasa Tosic for the assistance in the qRT-PCR experiments.

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Authors and Affiliations

  1. Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444a, PO Box 23, Belgrade, 11010, Serbia

    Jovana Jasnic-Savovic, Ana Kojic, Vlado Kovcic, Srdjan Boskovic, Aleksandra Nestorovic, Ljiljana Rakicevic, Dragica Radojkovic & Snezana Kojic

  2. Department of Neurology, Friedrich Baur Institute, Ludwig Maximilians University, Ziemssenstr. 1a, 80336, Munich, Germany

    Sabine Krause, Olivia Schreiber-Katz, Johannes G. Vogel, Benedikt G. Schoser & Maggie C. Walter

  3. Institute of Forensic Medicine, School of Medicine, University of Belgrade, Deligradska 31a, Belgrade, 11000, Serbia

    Slobodan Savic

  4. CRIBI, University of Padova, Via Ugo Bassi 58b, 35121, Padua, Italy

    Giorgio Valle & Georgine Faulkner

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  1. Jovana Jasnic-Savovic
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  2. Sabine Krause
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  3. Slobodan Savic
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  4. Ana Kojic
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  8. Ljiljana Rakicevic
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Correspondence to Snezana Kojic.

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All procedures performed in this study involving human participants were in accordance with the ethical standards of the Faculty of Medicine, University of Belgrade, 1964 Declaration of Helsinki and the Law on Health Care of the Republic of Serbia. For this type of study, formal consent is not required. All applicable international and national guidelines for the care and use of animals were followed.

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Jasnic-Savovic, J., Krause, S., Savic, S. et al. Differential expression and localization of Ankrd2 isoforms in human skeletal and cardiac muscles. Histochem Cell Biol 146, 569–584 (2016). https://doi.org/10.1007/s00418-016-1465-0

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