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EVOLUTION OF PROSTAGLANDINS: HAVE TELEOST FISH AN ACTIVE LIPOCALIN-TYPE PROSTAGLANDIN D SYNTHASE?
Autores:
VICTORIA GÓMEZ ABELLÁN
,
VICTORIANO MULERO MÉNDEZ
,
MªPILAR SEPULCRE CORTÉS
,
Grupos de investigación:
[GI/IMIB/C060/2011] Inmunidad, inflamación y cáncer
Comunicación:
Antecedentes:
Mammalian lipocalin-type Prostaglandin D synthase (L-PGDS) is a bifunctional protein with the ability of carrying lipophilic molecules and synthesizing PGD2. However studies about the activity of non mammalian L-PGDS revealed few or no activity for this molecule. The aim of this study is the identification of the different zebrafish L-PGDS isoforms and the analysis of their expression in larvae and adult zebrafish under control and infected conditions and to determine their enzymatic activity.
Métodos:
Sequence homology analysis was performed using BLAST of the NCBI nucleotide sequence database using the structure of human L-PGDS. Gene expression was analyzed with RT-PCR in zebrafish embryos during the first stages of development (0, 2, 7, 9, 24, 48 and 72 hpf). Zebrafish larvae were infected or not with Salmonella at 2 dpf, and the expression of the different L-PGDS isoforms was analyzed with RT-qPCR at 3, 6, 24 and 48 hpi. Adult zebrafish were infected or not with Vibrio anguillarum and 24 and 48 hpi the expression of the different L-PGDS isoforms were analyzed by RT-qPCR in head kidney.
Resultados:
In this study we have characterized three L-PGDS isoforms in the zebrafish gene database, (L-PGDSa, L-PGDSb, L-PGDSlip15) and have identified a new one in the chromosome 5 termed as L-PGDSnew5. All isoforms contain a lipocalin domain and the conserved folding patterns and three structurally conserved regions (SCRs) but, surprisingly, lacked the essential cysteine residue required for PGDS activity found in mammalian L-PGDS. The expression analysis in zebrafish embryos during the first stages of development showed that the transcripts of the four L-PGDS isoforms are maternally transferred since they were detected from the fertilization time. Furthermore, larvae infection with Salmonella result in increased mRNA levels of L-PGDSlip15 at all time tested. Similarly, adult zebrafish infection with Vibrio anguillarum was able to induce the transcript levels of L-PGDSlip15 in head kidney. In contrast bacterial infection decreased the mRNA levels of L-PGDSa, L-PGDSb, and L-PGDSnew5.
Conclusiones:
Our data suggests a key role of zebrafish L-PGDSlip15 in the response to bacterial infection. Further studies aimed at determining its enzymatic activity will through light into to the role of this enzyme in fish immunity.
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