Huc-Brandt Sylvaine, Hieu Nelson, Imberdis Thibaut, Cubedo Nicolas, Silhol Michelle, Leighton Patricia L A, Domaschke Thomas, Allison W Ted, Perrier Véronique, Rossel Mireille
Univ. Montpellier 2, Montpellier, F-34095 France; Inserm, U710, Montpellier, F-34095 France; EPHE, Paris, F-75007 France.
Boston University School of Medicine, Department of Biochemistry, Boston, MA 02118, United States of America.
PLoS One. 2014 Dec 1;9(12):e113331. doi: 10.1371/journal.pone.0113331. eCollection 2014.
Prion protein is involved in severe neurodegenerative disorders but its physiological role is still in debate due to an absence of major developmental defects in knockout mice. Previous reports in zebrafish indicate that the two prion genes, PrP1 and PrP2, are both involved in several steps of embryonic development thus providing a unique route to discover prion protein function. Here we investigate the role of PrP2 during development of a mechano-sensory system, the posterior lateral line, using morpholino knockdown and PrP2 targeted inactivation. We confirm the efficiency of the translation blocking morpholino at the protein level. Development of the posterior lateral line is altered in PrP2 morphants, including nerve axonal outgrowth and primordium migration defects. Reduced neuromast deposition was observed in PrP2 morphants as well as in PrP2-/- mutants. Rosette formation defects were observed in PrP2 morphants, strongly suggesting an abnormal primordium organization and reflecting loss of cell cohesion during migration of the primordium. In addition, the adherens junction proteins, E-cadherin and ß-catenin, were mis-localized after reduction of PrP2 expression and thus contribute to the primordium disorganization. Consequently, hair cell differentiation and number were affected and this resulted in reduced functional neuromasts. At later developmental stages, myelination of the posterior lateral line nerve was altered. Altogether, our study reports an essential role of PrP2 in collective migration process of the primordium and in neuromast formation, further implicating a role for prion protein in cell adhesion.
朊病毒蛋白与严重的神经退行性疾病有关,但其生理作用仍存在争议,因为敲除小鼠没有明显的发育缺陷。先前在斑马鱼中的报道表明,两个朊病毒基因PrP1和PrP2都参与胚胎发育的几个步骤,从而为发现朊病毒蛋白的功能提供了一条独特的途径。在这里,我们使用吗啉代敲低和PrP2靶向失活来研究PrP2在机械感觉系统——后侧线发育过程中的作用。我们在蛋白质水平上证实了翻译阻断吗啉代的有效性。PrP2 morphants中后侧线的发育发生改变,包括神经轴突生长和原基迁移缺陷。在PrP2 morphants以及PrP2 - / -突变体中观察到神经丘沉积减少。在PrP2 morphants中观察到玫瑰花结形成缺陷,强烈表明原基组织异常,并反映了原基迁移过程中细胞凝聚力的丧失。此外,在PrP2表达降低后,粘附连接蛋白E - 钙粘蛋白和β - 连环蛋白的定位错误,从而导致原基紊乱。因此,毛细胞的分化和数量受到影响,这导致功能性神经丘减少。在发育后期,后侧线神经的髓鞘形成发生改变。总之,我们的研究报告了PrP2在原基的集体迁移过程和神经丘形成中的重要作用,进一步暗示了朊病毒蛋白在细胞粘附中的作用。