Advances in Eicosanoid Research
Eicosanoids may therefore play a pivotal role in daphnid reproduction.
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A wealth of synthetic and natural chemicals may affect invertebrate reproduction through endocrine disruption with one of the best known examples being imposex masculinisation of female molluscs caused by exposure to tributyl tin TBT [ 13 ]. It is therefore important to understand eicosanoid biosynthesis in Daphnia , and invertebrates in general, to fully recognize the potential mode of action of endocrine disrupters and how they may affect natural invertebrate populations.
Recently, the genome of D. In the meantime genes identified in D. Here we present an overview of the putative eicosanoid biosynthesis in Daphnia based on annotation of genes from the D. Putative genes related to eicosanoid biosynthesis were identified on the D. Annotation of these genes were verified through BLAST searches performed against Swissprot protein and non-redundant protein sequence databases via the D. Invertebrate and vertebrate COX protein sequences were retrieved from GenBank [ 19 ] and Ensembl [ 20 ] for phylogenetic analysis.
Since no arthropod COX protein sequences were available that could aid the phylogeny with respect to D. After translating the nucleotide sequences to putative amino acids, it appeared that one H. The three remaining sequences constituted two slightly deviating amino acid sequences; DV differed from EH and DV The four H. Furthermore, the inferred amino acid sequences of the two P.
Note that these additional crustacean entries were EST derived, single pass sequenced, and therefore not guaranteed to be free from sequencing errors. The sequence COX dataset was aligned using ClustalW2 [ 21 ], manually edited after inspection and subsequently analysed using the Gblocks web-server [ 22 ] to pinpoint and remove unreliably aligned regions. This analysis allowed for gaps in the final alignment. ModelGenerator [ 23 ] was applied to obtain a model of sequence evolution gamma distribution with four rate categories using the Akaike Information Criterion.
Phylogenetic analysis used bootstrap replicates. Finally, the obtained topology was visualised using TreeView [ 26 ]. Table 1 shows the putative genes related to eicosanoid biosynthesis that were identified through searching the D.