Analysis of gelsolin expression pattern in developing chicken embryo reveals high GSN expression level in tissues of neural crest origin

Brain Structure and Function, Oct 2014

Gelsolin is one of the most intensively studied actin-binding proteins. However, in the literature comprehensive studies of GSN expression during development have not been performed yet in all model organisms. In zebrafish, gelsolin is a dorsalizing factor that modulates bone morphogenetic proteins signaling pathways, whereas knockout of the gelsolin coding gene, GSN is not lethal in murine model. To study the role of gelsolin in development of higher vertebrates, it is crucial to estimate GSN expression pattern during development. Here, we examined GSN expression in the developing chicken embryo. We applied numerous methods to track GSN expression in developing embryos at mRNA and protein level. We noted a characteristic GSN expression pattern. Although GSN transcripts were present in several cell types starting from early developmental stages, a relatively high GSN expression was observed in eye, brain vesicles, midbrain, neural tube, heart tube, and splanchnic mesoderm. In older embryos, we observed a high GSN expression in the cranial ganglia and dorsal root ganglia. A detailed analysis of 10-day-old chicken embryos revealed high amounts of gelsolin especially within the head region: in the olfactory and optic systems, meninges, nerves, muscles, presumptive pituitary gland, and pericytes, but not oligodendrocytes in the brain. Obtained results suggest that GSN is expressed at high levels in some tissues of ectodermal origin including all neural crest derivatives. Additionally, we describe that silencing of GSN expression in brain vesicles leads to altered morphology of the mesencephalon. This implies gelsolin is crucial for chicken brain development.

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Analysis of gelsolin expression pattern in developing chicken embryo reveals high GSN expression level in tissues of neural crest origin

Antonina Joanna Mazur 0 1 Gabriela Morosan-Puopolo 0 1 Aleksandra Makowiecka 0 1 Maria Malicka-Baszkiewicz 0 1 Dorota Nowak 0 1 Beate Brand-Saberi 0 1 0 G. Morosan-Puopolo B. Brand-Saberi Department of Anatomy and Molecular Embryology, Ruhr University of Bochum , Bochum, Germany 1 A. J. Mazur (&) A. Makowiecka M. Malicka-Baszkiewicz D. Nowak Department of Cell Pathology, Faculty of Biotechnology, University of Wroclaw , ul. Joliot-Curie 14a, 50-383 Wrocaw, Poland Gelsolin is one of the most intensively studied actin-binding proteins. However, in the literature comprehensive studies of GSN expression during development have not been performed yet in all model organisms. In zebrafish, gelsolin is a dorsalizing factor that modulates bone morphogenetic proteins signaling pathways, whereas knockout of the gelsolin coding gene, GSN is not lethal in murine model. To study the role of gelsolin in development of higher vertebrates, it is crucial to estimate GSN expression pattern during development. Here, we examined GSN expression in the developing chicken embryo. We applied numerous methods to track GSN expression in developing embryos at mRNA and protein level. We noted a characteristic GSN expression pattern. Although GSN transcripts were present in several cell types starting from early developmental stages, a relatively high GSN expression was observed in eye, brain vesicles, midbrain, neural tube, heart tube, and splanchnic mesoderm. In older embryos, we observed a high GSN expression in the cranial ganglia and dorsal root ganglia. A detailed analysis of 10-day-old chicken embryos revealed high amounts of gelsolin especially within the head region: in the olfactory and optic systems, meninges, nerves, muscles, presumptive pituitary gland, and pericytes, but not oligodendrocytes in the brain. Obtained results suggest that GSN is expressed at high levels in some tissues of ectodermal origin including all neural crest derivatives. Additionally, we describe that silencing of GSN expression in brain vesicles leads to altered morphology of the mesencephalon. This implies gelsolin is crucial for chicken brain development. - Actins, abundantly expressed in all animal cell types, are capable of forming polymers and take part in several cellular processes such as cell motility, chemoattractantcontrolled (or directed) migration, trafficking of cellular organelles and chromosomes, junction formation, mitosis, transcription, and muscle contraction (Perrin and Ervasti 2010). Consequently, it is understandable that actin dynamics have to be strictly controlled. There are more than 100 Actin-binding proteins (ABPs), which regulate actin polymerization/depolymerization and involvement in several cellular processes (Winder and Ayscough 2005). Although it was shown that actin is indispensable for mammalian embryonic development (Shawlot et al. 1998), not much is known about the role of ABPs in vertebrate embryo development. There are, however, some exceptions. For instance, it has been shown that thymosin beta4 plays a role in brain development of chicken embryos (Wirsching et al. 2012) and that it is expressed in the neural tube and brain vesicles, the heart, blood vessels, and feathers (Dathe and Brand-Saberi 2004). The presence of thymosin beta4 mRNA was strongly manifested in neural tissues like the neural tube and dorsal root ganglia. These results are similar to those obtained by researchers, who studied thymosin beta4 expression in mouse embryos (Gomez-Marquez et al. 1996). In contrast, another thymosin beta family member, thymosin beta15avian is asymmetrically expressed in Hensens node and could thus be involved in left/right axis formation; it has been described to have a function in myogenesis in chicken embryos (Chankiewitz et al. 2014). Other G-actin-binding proteins belong to the ADF/cofilin family. Actin depolymerizing factor (ADF) seems to be dispensable during mouse embryo development (Gurniak et al. 2005), but n-cofilin is crucial for migration of cells derived from the paraxial mesoderm. On the other hand, decreased expression of non-muscle cofilin (n-cofilin) in murine preimplantation embryos is important for compaction during blastocyst formation (Ma et al. 2009). Here, we focus on the gelsolin expression pattern in developing chicken embryos. Gelsolin is coded by one gene (GSN); however, its expression yields several protein isoforms. In humans, the existence of three isoforms: plasma (isoform a, 86 kDa), cytoplasmic (isoform b, 81 kDa), and brain (isoform c, 82 kDa) is well documented. Furthermore, the presence of some other isoforms can be predicted. Gelsolin, a Ca2?, phosphatidylinositol 4,5-biphosphate (PIP2) and pH-dependent six-domain (G1 G6) protein, severs actin filaments, caps the barbed ends of actin filaments and under certain conditions nucleates actin monomers (Mannherz et al. 2010; Li et al. 2012; Nag et al. 2013). However, gelsolin seems to have more functions than severing actin filame (...truncated)


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Antonina Joanna Mazur, Gabriela Morosan-Puopolo, Aleksandra Makowiecka, Maria Malicka-Błaszkiewicz, Dorota Nowak, Beate Brand-Saberi. Analysis of gelsolin expression pattern in developing chicken embryo reveals high GSN expression level in tissues of neural crest origin, Brain Structure and Function, 2016, pp. 515-534, Volume 221, Issue 1, DOI: 10.1007/s00429-014-0923-5