Brain Research Bulletin 90 (2013) 118–126 Contents lists available at SciVerse ScienceDirect Brain Research Bulletin jo u rn al hom epa ge : www.el sevier.com/locate/brainresbull Research report The zebrafish homologue of Parkinson’s disease ATP13A2 is essential for embryonic survival Tomás Lopes da Fonseca a,d , Ana Correia a,b , Wiebren Hasselaar c , Herma C. van der Linde c , Rob Willemsen c,1 , Tiago Fleming Outeiro a,b,d,∗,1 a Cell and Molecular Neuroscience Unit, Instituto de Medicina Molecular, Lisboa, Portugal b Instituto de Fisiologia, Faculdade de Medicina da Universidade de Lisboa, Av. Professor Egas Moniz, 1649-028 Lisboa, Portugal c Dept of Clinical Genetics, Erasmus MC, Rotterdam, The Netherlands d Department of Neurodegeneration and Restorative Research, Center for Molecular Physiology of the Brain, University Medizin Göttingen, Waldweg 33, 37073 Göttingen, Germany a r t i c l e i n f o Article history: Received 4 July 2012 Received in revised form 16 September 2012 Accepted 30 September 2012 Available online 31 October 2012 Keywords: ATP13A2 Zebrafish Parkinson’s disease Development Kufor-Rakeb syndrome a b s t r a c t ATP13A2 is a lysosome-specific transmembrane ATPase protein of unknown function. This protein was initially linked to Kufor-Rakeb syndrome where it is absent or mutated. More recently, point mutations in ATP13A2 were linked to familial cases of Parkinson’s disease. Zebrafish is commonly used as a vertebrate model for the study of different neurodegenerative diseases and has homologues of several Parkinson’s disease associated proteins. Here, we describe for the first time the zebrafish homologue of human ATP13A2, demonstrating the homology between the protein sequences, which supports a conserved biological role. Furthermore, the spatial pattern of protein expression was studied and the lethality of the knockdown of ATP13A2 suggests it plays a crucial role during embryonic development. Our findings bring new insight into the biology of ATP13A2 and open novel opportunities for its study using zebrafish as a model organism. © 2012 Elsevier Inc. All rights reserved. 1. Introduction ATP13A2 is a lysosomal membrane protein that belongs to the P5 subfamily of P-type transport ATPases, a transporter family of 5 proteins (ATP13A1–5) with unknown substrates (Schultheis et al., 2004). ATP13A2 was first associated to Kufor-Rakeb syndrome (Najim al-Din et al., 1994), a disease characterized by sub-acute, juvenile-onset, levodopa-responsive parkinsonism with signs of dementia (Williams et al., 2005). Recently, ATP13A2 was associ- ated with Parkinson’s disease (PD), since it was found mutated in several early onset PD cases (Ramirez et al., 2006; Di Fonzo et al., 2007; Ning et al., 2008; Djarmati et al., 2009; Behrens et al., 2010; Crosiers et al., 2011; Santoro et al., 2011). The pathological hallmarks of PD are the degeneration of dopaminergic neurons and the accumulation of cytoplasmic pro- tein inclusions named Lewy bodies that can be found in the remaining surviving neurons (Spillantini et al., 1997). Neverthe- less, the molecular mechanisms underlying PD pathology are still ∗ Corresponding author at: Department of Neurodegeneration and Restorative Research, University Medizin Göttingen, Waldweg 33, 37073 Göttingen, Germany. Tel.: +49 5513913544; fax: +351 21 799 9436. E-mail address: touteiro@gmail.com (T.F. Outeiro). 1 These two authors contributed equally to this article. unclear. Lewy bodies are mainly constituted by alpha-synuclein, which was the first protein to be associated with familial cases of the disease (Mezey et al., 1998). ATP13A2 was described to be in the same network of alpha-synuclein, since it was able to overcome the toxicity induced by overexpression of the latter protein (Gitler et al., 2009). More recently, a role in mitochondrial regulation through macroautophagy was found for ATP13A2, where knockdown (kd) of the gene promoted an increase of mitochondria fragmentation and reactive oxygen species (ROS) production (Gusdon et al., 2012) with the mutation of the gene impairing mitochondria function (Grunewald et al., 2012). Moreover, it was shown that ATP13A2 expression is strongly regulated by hypoxia signalling (Xu et al., 2012). This data suggests a novel interaction between two crucial cellular systems: the degradation and the mitochondrial system, both strongly linked to PD pathology. Several model organisms have been used to investigate the underlying pathobiology of PD (Recchia et al., 2004; Melrose et al., 2006). Unfortunately, none has been able to fully recapitulate all central aspects of the disease, justifying the need for developing new models. Danio rerio, commonly designated as zebrafish, is a vertebrate organism that is widely used as a model to study the molecu- lar basis of several human disorders due to the high degree of 0361-9230/$ – see front matter © 2012 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.brainresbull.2012.09.017