Instrumentos de Medición

Overall, we demonstrate that Shp2 plays a crucial role in zebrafish development. Genetic duplication of ptpn11 allowed us to study gene dosage effects and the role of Shp2a and Shp2b at various stages of development (Chapter 4). Moreover, we show that both NS and LS Shp2 cause hypo- and hyperphosphorylation of Fer and PZR, respectively compared to WT. Genetic interaction studies indicated a role for reduced signaling by Fer in the etiology of NS and LS (Chapter 5). Functional studies showed indications for enhanced signaling complex formation of NS and LS Shp2 with PZR and Src (Chapter 6). Changes in protein-protein interactions are an important contributor to NS and LS. Both NS and LS Shp2 can lead to GOF phenotypes, showing enhanced Erk phosphorylation at the end of gastrulation in zebrafish. Both NS and LS lead to early heart defects in a similar way as well (Chapter 7). In addition, we identified mutations in an extracellular protein, A2ml1, to be associated with Noonan-like phenotypes (Chapter 8). Taken together, our findings show that NS and LS Shp2 share many mechanistic and developmental features, and that these disease associated proteins should not only be viewed as “activating” or “inactivating”. Using a versatile in vivo model, we show that these proteins should rather be viewed in a network of interacting proteins that gives rise to similar developmental defects. Overall, we used zebrafish embryos as a model to study the etiology of NS and LS at the genetic, molecular, proteomic and organismal level.

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Appendix