Haplotype Analysis of the First A4V-SOD1 Spanish Family: Two Separate Founders or a Single Common Founder?

A4V; SOD1; Esclerosi lateral amiotròficaA4V; SOD1; Esclerosis lateral amiotróficaA4V; SOD1; Amyotrophic lateral sclerosisDespite the genetic heterogeneity reported in familial amyotrophic lateral sclerosis (ALS) (fALS), Cu/Zn superoxide-dismutase (SOD1) gene mutations are the second most common cause of the disease, accounting for around 20% of all families (ALS1) and isolated sporadic cases (sALS). At least 186 different mutations in the SOD1 gene have been reported to date. The possibility of a single founder and separate founders have been investigated for D90A (p.D91A) and A4V (p.A5V), the most common mutations worldwide. High-throughput single nucleotide polymorphism genotyping studies have suggested two founders for A4V (one for the Amerindian population and another for the European population) although the possibility that the two populations are descended from a single ancient founder cannot be ruled out. We used 15 genetic variants spanning the human chromosome 21 from the SOD1 gene to the SCAF4 gene, comparing them with the population reference panels, to demonstrate that the first A4V Spanish pedigree shared the genetic background reported in the European population.This study has been supported by Instituto de Salud Carlos III (grant numbers PIS-FEDER PI16/01673 and PI19/00593). JG and JV-T are the recipients of grant 2017SGR00939 from Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR) of the Generalitat de Catalunya

Regulación de la movilidad celular por Profilina 1

Mi proyecto de tesis se basaba en el estudio del papel de profilina 1 en la formación de lamelas, para ello generamos una proteína recombinante y transducible, con el objetivo de poder modificar los niveles endógenos de profilina. Objetivos: i-caracterización bioquímica los tres sitios de union conocidos de la proteína de transducción, el sitio de unión a fosfo-inocitoles (PIP), el de unión a actina (Ac) y el de unión a poli-prolinas (PLP). ii-estudio de la polimerización in-vitro de actina - PTD4-Profilina1 iii-estudio de las proteínas componentes de lamelas inducidas por PTD4-Profilina1. Plan de trabajo: i-Para comprobar la funcionalidad los 3 sitios de unión fueron necesarias las primeras 6 semanas, ya que en primer lugar había que expresar y purificar el peptido Srv2, necesario para el ensayo de PLP. En segundo lugar, se obtuvieron los datos de las concentraciones adecuadas de lípidos para el ensayo de fosfo-inocitoles y por ultimo, se purifico la actina necesaria para el ensayo de unión a actina. Una vez establecida la funcionalidad de la proteína, se procedió a: ii-el estudio de polimerización in-vitro, que llevo 2 semanas. Demostrando que in-vitro era capaz de inhibir la polimerización de una manera similar a la endógena. Una vez terminados estos ensayos, se procedio a: iii-la caracterización inmunohistoquímica de las proteínas componentes de la lamela que fue llevado a cabo en 4 semanas. Para ello se usaron anticuerpos contra: alfa-actinina, talina, vinculina, ENA/Vasp y paxillina. Conclusiones: i-las propiedades bioquímicas de la PTD4-Profilina1 son similares a las de la profilina endógena. ii-los estudios de polimerización indican que la polimerización se produce de manera similar a la endogena. iii-los ensayos de inmunohistoquímica sugieren que, talina esta ausente y que las demás están presentes aunque en menor concentración y con otra distribución comparadas con los controles.My thesis project is based in the study of roll of profilin lamella formation. To achieve it, we developed a recombinant and transducible protein called PTD4-Pfn1. With this protein, we are able to modified the intracellular protein levels in a fisiological range . During the time between the grant submission and the beginning of the stay we developed in the objectives, and for an explicit petition of the researcher director of the acceptor center, we changed the objectives in the following: Objetives i-biochemical characterization of the three known binding sites of the of the PTD4-Profilin1 protein transduction, the phosphor-inocitol, actin and poly-prolines binding sites. ii-in-vitro study of the actin polymerization in presence of PTD4-Profilin1. iii-study of the components proteins in the formation of the lamellas PTD4-Profilin induced. Planning i-To test the three binding sites 6 weeks were neccesary. In these weeks we purified the peptides necessary to test the poly-proline binding sites, the Srv2. We also tested the best lipids concentration to the phosphor-inocitol binding assay. Finally, we prepared the actin necessary to the actin binding assay. Once established the protein functionality, we proceed to the following experiments: ii- the in vivo actin polylerization assay was performed in 2 weeks. And: iii-the inmuno-hystochemical characterization of the proteins of the PTD4-Profilin induced was performed in 4 weeks. For this propouse ee used antibodies against alfa-actinin, talin, vinculin, ENA/Vasp y paxillin. Conlusions i-the biochemical properties of PTD4-Profilin1 are similars to the endogen profilin. ii-Lo studies suggest that the actin polymerization induced by PTD4-Profilin1 is similar to the promoted by the endogen profilin1 iii-the hystochemical assays suggest that talin is absent and the others, alfa-actinin, vinculin, ENA/Vasp and paxillin are present but il low concentration and in a different distribution