Centromeres are an important part of chromosomes which direct chromosome segregation during cell division. Their modifications can therefore explain the unusual mitotic and meiotic behaviour of certain chromosomes, such as the germline-restricted chromosome (GRC) of songbirds. This chromosome is eliminated from somatic cells during early embryogenesis and later also from male germ cells during spermatogenesis. Although the mechanism of elimination is not yet known, it is possible that it involves a modification of the centromeric sequence on the GRC, resulting in problems with the attachment of this chromosome to the mitotic or meiotic spindle and its lagging during anaphase, which eventually leads to its elimination from the nucleus. However, the repetitive nature and rapid evolution of centromeres make their identification and comparative analysis across species and chromosomes challenging. Here, we used a combination of cytogenetic and genomic approaches to identify the centromeric sequences of two closely related songbird species, the common nightingale (Luscinia megarhynchos) and the thrush nightingale (L. luscinia). We found a 436-bp satellite repeat present in the centromeric regions of all regular chromosomes (i.e., autosomes and sex chromosomes), making it a strong candidate for the centromeric repeat. This centromeric repeat was highly similar between the two nightingale species. Interestingly, hybridization of the probe to this satellite repeat on meiotic spreads suggested that this repeat is missing on the GRC. Our results indicate that the change of the centromeric sequence may underlie the unusual inheritance and programmed DNA elimination of the GRC in songbirds.

Bonn Institute for Organismal Biology Animal Biodiversity University of Bonn Bonn Germany

Centre for Molecular Biodiversity Research Leibniz Institute for the Analysis of Biodiversity Change Bonn Germany

Department of Behavioural Ecology Institute of Environmental Biology Faculty of Biology Adam Mickiewicz University Poznan Poland

Department of Organismal Biology Systematic Biology Science for Life Laboratory Uppsala University Uppsala Sweden

Department of Zoology Faculty of Science Charles University Prague Czech Republic

Department of Zoology Faculty of Science Palacký University Olomouc Czech Republic

Genomics Core Facility European Molecular Biology Laboratory Heidelberg Germany

Institute for Environmental Studies Faculty of Science Charles University Prague Czech Republic

Institute of Vertebrate Biology Czech Academy of Sciences Studenec Czech Republic

Laboratory of Genomics and Bioinformatics Institute of Molecular Genetics Czech Academy of Sciences Prague Czech Republic

Laboratory of Non Mendelian Evolution Institute of Animal Physiology and Genetics Czech Academy of Sciences Liběchov Czech Republic

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