Analysis of 14-3-3 family member function in Xenopus embryos by microinjection of antisense morpholino oligos.

Abstract

The 14-3-3 intracellular phosphoserine/threonine-binding proteins are adapter molecules that regulate signal transduction, cell cycle, nutrient sensing, apoptotic, and cytoskeletal pathways. There are seven 14-3-3 family members, encoded by separate genes, in vertebrate organisms. To evaluate the role of individual 14-3-3 proteins in vertebrate embryonic development, we utilized an antisense morpholino oligo microinjection technique in Xenopus laevis embryos. By use of this method, we showed that embryos lacking specific 14-3-3 proteins displayed unique phenotypic abnormalities. Specifically, embryos lacking 14-3-3 tau exhibited gastrulation and axial patterning defects, but embryos lacking 14-3-3 gamma exhibited eye defects without other abnormalities, and embryos lacking 14-3-3 zeta appeared completely normal. These and other results demonstrate the power and specificity of the morpholino antisense oligo microinjection technique.