Previous studies have also indicated that myosin-Va is found in synaptic vesicle preparations and forms stable complexes between synaptic vesicle membrane proteins (Mani et al., 1994 and Prekeris and Terrian, 1997). In the vertebrate brain, 5–15% of the total zinc is concentrated in synaptic vesicles

(Frederickson, 1989 and Frederickson and Moncrieff, 1994), which has been studied using the Neo-Timm method (Babb et al., 1991). Moreover, zinc serves as an endogenous neuromodulator of several important receptors, including N-methyl-d-aspartate (NMDA) ( Smart et al., 1994). Functional studies of honey bee myosin-Va have not been carried out until now. In this study, we addressed the effects of intracerebral injections of melittin PI3K inhibitor and NMDA on the honey bee. Melittin is a polypeptide present in bee venom (Habermann, 1972) and a potent calmodulin

antagonist (Steiner et al., 1986). Calmodulin is the most extensively studied member of the intracellular calcium-binding proteins, which includes myosin-Va. Additionally, NMDA is a glutamate-gated ion channel agonist present in both mammals and insects (Paoletti and Neyton, 2007). The selleck screening library NMDA receptor is involved in delayed neuronal death (Choi, 1988) and excitatory synaptic transmission in the central nervous system, which results in learning and memory (Albensi, 2007). A critical role of the NMDA receptor was recently demonstrated in olfactory learning and memory in Drosophila melanogaster ( Xia et al.,

2005) and A. mellifera ( Locatelli et al., 2005 and Si et al., 2004). The aims of this study were to elucidate some of the biochemical properties and the distribution of myosin-Va and to describe the expression patterns of molecular motors and SNARE proteins in the honey bee (A. mellifera L.) brain. Moreover, we evaluated the alterations in myosin-Va expression after intracerebral injections of melittin and NMDA. Rabbit affinity-purified polyclonal antibodies were used in this study. Anti-chicken brain myosin-Va (α-myosin-Va) head domain recombinant protein (Espreafico et al., 1992 and Suter et al., 2000), anti-pig myosin-VI (α-myosin-VI) tail fusion protein (Hasson and Mooseker, 1994) and anti-myosin-IXb Glutathione peroxidase heavy chain tail domain recombinant protein (Post et al., 1998) were all from the Mooseker Laboratory (Yale University, New Haven, CT, USA). Anti-rabbit myosin-IIb (α-myosin-IIb) was produced in the Larsons Laboratory (USP, Ribeirão Preto, SP, Brazil). The dynein light chain (α-DYNLL1/LC8) antibody was generated against the Chlamydomonas LC8 recombinant protein ( King et al., 1996). Mouse monoclonal antibodies used included anti-cytoplasmic dynein intermediate chain IC74 (α-DIC; Chemicon International Inc.