DECIPHERING CNBP CELLULAR AND MOLECULAR FUNCTION DURING CRANIAL NC DEVELOPMENT WITH A NOVEL MORPHOLINO TECHNIQUE

WEINER, AM; KELSH, RN; CALCATERRA, N.B.

Santa Cruz

Congreso; 5th International Meeting of the Latin American Society of Developmental Biology (LASDB).; 2010

LASDB

DECIPHERING CNBP CELLULAR AND MOLECULAR FUNCTION DURING CRANIAL NC DEVELOPMENT WITH A NOVEL MORPHOLINO TECHNIQUE Andrea M.J. Weiner1, Robert N. Kelsh2, and Nora B. Calcaterra1. 1Instituto de Biología Molecular y Celular de Rosario, Consejo Nacional de Investigaciones Científicas y Técnicas - Área Biología, Dpto. de Ciencias Biológicas, Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina. 2Centre for Regenerative Medicine, Developmental Biology Programme, Department of Biology and Biochemistry, University of Bath, Bath, UK. weiner@ibr.gov.ar Several issues need to be well thought-out when functionally studying maternally inherited genes during development. Without considering the use of mutant or transgenic animals, some loss-of-function methods properly knockdown an mRNA expression while others block protein function by the expression of dominant negative isoforms. However, it is known that those methods do not affect the maternally inherited protein pool and also the dominant negative isoforms are degraded or titled as development advances, being unable to reduce newly synthesized proteins. In this work, we report that the use of a morpholino that specifically impairs splicing of cellular nucleic acid binding protein (CNBP) pre-mRNA intron 2 leads to the synthesis of shorter protein isoforms. Furthermore, these appear to act as dominant negatives, enabling us to elucidate CNBP function during NC development. Our studies demonstrate that CNBP loss-of-function adversely affects formation and survival of a subpopulation of the cranial NC cells, leading to reduction/loss of selected pharyngeal and craniofacial cartilaginous structures in the developing zebrafish. These defects result from disruption of CNBP roles in both NC cell survival and specification. The strategy used in this work may be an interesting experimental tool for studying the role of other maternally inherited proteins during embryonic development.