PROTEASOME INHIBITORS IMPROVE TREACHER COLLINS SYNDROME (TCS) IN A ZEBRAFISH (Danio rerio) MODEL.

LORENZATTI, AGUSTÍN; GIL ROSAS, MAUCO; CALCATERRA, NORA; COUX, GABRIELA

Buenos Aires

Congreso; I Reunion Conjunta de Sociedades de Biociencias 2017; 2017

TCS is a congenital disease characterized by defects in the craniofacial skeleton and absence of mental alterations. We modeled TCS in D. rerio embryos through the microinjection of Morpholino oligonucleotides blocking the translation of the ortholog of the causative gene (TCOF1). Cnbp, a protein required for proper craniofacial development, was detected in lower levels (without changes in its mRNA expression) in TCS-like embryos. As Cnbp degradation is carried out through the proteasomal pathway, we tested if proteasome inhibitors (MG132 and Bortezomib) were able to ameliorate cranial skeleton malformations in TCS. Two-cell embryos were injected with control Morpholino (C) or tcof1 translation blocking Morpholino (TC). At 6 hours post fertilization (hpf) embryos were exposed to MG-132 (5 µM), Bortezomib (0.5 µM) or vehicle for 18 hours. Embryo viability was not affected by any treatment. Control embryos did not show any effect on cranial cartilages induced by MG132, bortezomib or the vehicle, and were pooled together as control group. Cnbp protein level measured by western-blot in total extracts prepared from 24 hpf treated specimens recovered under both treatments, although not to control levels. Cranial cartilages measurements were performed in 4 days post-fertilization larvae stained with Alcian Blue by using the ImageJ software. In arbitrary units, Meckel length: C:100.0 ±0.7, TC: 89.3±3.1*, TC+Bort: 98.7266±2.0, TC+MG132: 91.5795±2.8, Ceratohyal angle: C: 99.4±1.3, TC: 157.3±14.9*, TC+Bort: 103.0±4.6, TC+MG132: 124.3±10.7, p