title: A functional Bucky ball-GFP transgene visualizes germ plasm in living zebrafish.
abstract: In many animals, the germline is specified by maternal RNA-granules termed germ plasm. The correct localization of germ plasm during embryogenesis is therefore crucial for the specification of germ cells. In zebrafish, we previously identified Bucky ball (Buc) as a key regulator of germ plasm formation. Here, we used a Buc antibody to describe its continuous germ plasm localization. Moreover, we generated a transgenic Buc-GFP line for live imaging, which visualizes germ plasm from its assembly during oogenesis up to the larval stages. Live imaging of Buc-GFP generated stunning movies, as they highlighted the dynamic details of germ plasm movements. Moreover, we discovered that Buc was still detected in primordial germ cells 2 days after fertilization. Interestingly, the transgene rescued buc mutants demonstrating genetically that the Buc-GFP fusion protein is functional. These results show that Buc-GFP exerts all biochemical interactions essential for germline development and highlight the potential of this line to analyze the molecular regulation of germ plasm formation.
authors.affiliation: Institut für Entwicklungsbiochemie, GZMB, Universitätsmedizin Göttingen, Georg-August-Universität, Göttingen, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Riemer
authors.affiliation: Laboratory of Metabolism, Department of Internal Medicine Specialties, Faculty of Medicine, University of Geneva, Switzerland.
authors.forename: Franck
authors.initials: F
authors.lastname: Bontems
authors.affiliation: Institut für Entwicklungsbiochemie, GZMB, Universitätsmedizin Göttingen, Georg-August-Universität, Göttingen, Germany.
authors.forename: Pritesh
authors.initials: P
authors.lastname: Krishnakumar
authors.affiliation: Institut für Entwicklungsbiochemie, GZMB, Universitätsmedizin Göttingen, Georg-August-Universität, Göttingen, Germany.
authors.forename: Jasmin
authors.initials: J
authors.lastname: Gömann
authors.affiliation: Institut für Entwicklungsbiochemie, GZMB, Universitätsmedizin Göttingen, Georg-August-Universität, Göttingen, Germany. Electronic address: roland.dosch@med.uni-goettingen.de.
authors.forename: Roland
authors.initials: R
authors.lastname: Dosch
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D000906:Antibodies / Q000378:metabolism; D005854:Germ Cells / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics*; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D009866:Oogenesis; D011993:Recombinant Fusion Proteins / Q000235:genetics; D015027:Zebrafish / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
chemical_list: D000906:Antibodies; D011993:Recombinant Fusion Proteins; D029961:Zebrafish Proteins; C541253:buc protein, zebrafish; D049452:Green Fluorescent Proteins
keywords: Balbiani body; Bucky ball; Germline development; Primordial germ cell; Transgene; Vasa