In Vivo analysis of 14-3-3 proteins

Abstract

The 14-3-3 proteins are small acidic cytosolic proteins abundant in the nervous system, but without a well established physiological function. Putative functions assigned to these proteins include regulation of Ca²⁺ exocytosis, and activity modulation of the kinases PKC, Raf and Bcr. The Drosophila 14-3-3[] gene (leonardo) is expressed preferentially in mushroom bodies, neural centers for insect learning and memory. Lesions in leonardo that significantly decrease the level of protein in the mushroom bodies precipitate deficits in negatively reinforced olfactory learning. The homozygous mutants in the leo gene are lethal. This is an in vivo study of 14-3-3[] function by conditional rescue of lethality of homozygous leo mutants by regulated transgene expression. This protocol can be utilized to study the role of genes involved in vital functions and in behavior. The rescue parameters and behavioral phenotype of rescued animals are reported herein. Furthermore, rescue of the learning phenotype achieved by heat shock induction of the transgene and examination of potential functional differences between leo I and leo II are reported.