RESUMEN
BACKGROUND: Chemoreception is vitally important for all animals, yet little is known about the genetics of chemoreception in aquatic organisms. The keystone species Daphnia pulex, a well known crustacean, is the first aquatic invertebrate to have its genome sequenced. This has allowed us the initial investigation of chemoreceptor genes in an aquatic invertebrate, and to begin the study of chemoreceptor evolution across the arthropod phylum. RESULTS: We describe 58 Grs (gustatory receptors), belonging to the insect chemoreceptor superfamily, which were identified bioinformatically in the draft genome of the crustacean waterflea Daphnia pulex. No genes encoding proteins similar to the insect odorant receptors (Ors) were identified. These 58 Grs form 3 distinctive subfamilies of 37, 12, and 5 genes, as well as a highly divergent singleton (Gr58). In addition, Grs55-57 share distinctive amino acid motifs and cluster with the sugar receptors of insects, and may illuminate the origin of this distinctive subfamily. ESTs, tiling array, and PCR amplification results support 34 predicted gene models, and preliminary expression data comparing the sexes indicates potential female-biased expression for some genes. CONCLUSION: This repertoire of 58 chemoreceptors presumably mediates the many chemoperception abilities of waterfleas. While it is always possible that the entire Or gene lineage was lost at some point in the history of Daphnia pulex, we think it more likely that the insect Or lineage is indeed a relatively recently expanded gene lineage concomitant with the evolution of terrestriality in the insects or their hexapod ancestors.
Asunto(s)
Daphnia/genética , Receptores de Superficie Celular/genética , Animales , Femenino , Regulación de la Expresión Génica , Variación Genética , Proteínas de Insectos/genética , Masculino , Filogenia , Receptores Odorantes/genéticaRESUMEN
Standing square-wave chronoamperometry (SSWCA) was applied to the analysis of the microfluid flow generated by the movement of the appendages of the Crustacea Daphnia. This novel approach provided for the first time real-time assessment and analysis of the breathing rate/fluid flow of individual organisms. An electrochemical tracer was delivered into the fluid inflow of the organism and a carbon fiber microelectrode placed in the fluid outflow's path. The variation of the net concentration/flux of the electroactive tracer, dopamine, at the electrode surface was measured with SSWCA. The observed chronoamperometric peaks (with fine structure) of the outflow are seen as a direct representation of appendage movement and, too, the workings and responses of the organism to its environment, e.g., external stimuli such as food or chemicals. It was concluded that SSWCA follows primarily the variation of the convective component of the Nernst-Plank equation for flux and, to lesser extent, diffusion and migration. In this work, SSWCA can clearly be used to monitor changes in the Daphnia-generated fluid outflow on a different time scale than was previously possible. This new application of SSWCA is faster and likely more accurate than using high-speed video.