RESUMO
To clarify the role of phospholipase C (PLC) in insect olfactory transduction, we have undertaken its molecular identification in the moth Spodoptera littoralis. From the analysis of a male antennal expressed sequence tag library, we succeeded in cloning a full-length cDNA encoding a PLC that belongs to the cluster of PLC-beta subtypes. In adult males, the PLC-beta transcript was located predominantly in brain and antennae where its presence was detected in the olfactory sensilla trichodea. Moreover, PLC-beta was expressed in antennae at the beginning of the pupal stage, then reached a maximum at the end of this stage and was maintained at this level during the adult period. Taken together, these results provided molecular evidence for the putative participation of a PLC-beta in signaling pathways responsible for the establishment and the functioning of insect olfactory system.
Assuntos
Encéfalo/metabolismo , Condutos Olfatórios/metabolismo , Neurônios Receptores Olfatórios/fisiologia , Fosfolipase C beta/fisiologia , Spodoptera/enzimologia , Animais , DNA Complementar/genética , Masculino , Mariposas , Fosfolipase C beta/genética , Proteínas Recombinantes/farmacologia , Spodoptera/genéticaRESUMO
Signal inactivation is a crucial step in the dynamic of olfactory process and involves various Odorant-Degrading Enzymes. In the silkworm Bombyx mori, one of the best models for studying olfaction in insects, the involvement of an antennal-specific aldehyde oxidase in the degradation of the sex pheromone component bombykal has been demonstrated over the three past decades by biochemical studies. However, the corresponding enzyme has never been characterized at the molecular level. Bioinformatic screening of B. mori genome and molecular approaches have been used to isolate several candidate sequences of aldehyde oxidases. Two interesting antennal-expressed genes have been further characterized and their putative functions are discussed in regard to their respective expression pattern and to our knowledge on aldehyde oxidase properties. Interestingly, one gene appeared as specifically expressed in the antennae of B. mori and associated in males with the bombykal-sensitive sensilla, strongly suggesting that it could encode for the previously biochemically characterized enzyme.
Assuntos
Aldeído Oxidase/genética , Bombyx/enzimologia , Genes de Insetos , Feromônios/metabolismo , Olfato/genética , Aldeído Oxidase/química , Aldeído Oxidase/classificação , Sequência de Aminoácidos , Animais , Bombyx/genética , Feminino , Genoma de Inseto/genética , Masculino , Dados de Sequência Molecular , FilogeniaRESUMO
The non-steroidal ecdysone agonist, RH-5992, exhibits ecdysteroid activities in vivo as well as in vitro more effectively than 20-hydroxyecdysone (20E). Using the IAL-PID2 cells derived from imaginal wing discs of last larval instar of Plodia interpunctella, we investigated the action of RH-5992 in the control of cell growth. Its effects on the proliferative activity of IAL-PID2 cells, the induction level in G2/M arrest and on the expression rate of Plodia B cyclin (PcycB), ecdysone B1-isoform (PIEcR-B1) and Ultraspiracle-2 isoform (PIUSP-2) were examined. From these cellular and molecular assays, our results brought evidence that RH-5992, like 20E, induced an inhibition on cell proliferation by blocking IAL-PID2 cells in G2/M phase. Moreover, this G2/M arrest was preceded by a decrease in the expression level of PcycB and a high induction of PIEcR-B1, PIUSP-2 mRNAs. Dose-response experiments revealed that RH-5992 was even more potent than 20E. On these parameters, we therefore suggest that the differential observed in the expression level of USP and EcR by RH-5992 and 20E could contribute to the difference observed for the biological potency of these two compounds.