Project description:Previous works in the framework of EU ARRAINA Project evidenced a pro-inflammatory condition in gilthead sea bream (Sparus aurata) fed extremely low fish meal/fish oil diets, and this effect was mostly reversed by butyrate supplementation. The hypothesis of work is that these nutritionally-mediated changes can be extensive to intestinal mucus proteome and gut microbiota, which in turn could modify disease outcome.s If so, the prevalence and progression of the disease might be also modified by diet composition and feed additives. Gilthead sea bream fingerlings were fed with control and experimental diets formulated by BioMar until two year-old. FM was added at 25% in the control diet (D1) and at 5% in the other three diets (D2-D4). Added oil was either FO (D1 control diet) or a blend of vegetable oils, replacing the 58% (D2) and the 84% (D3-D4 diets) of FO. A commercial sodium butyrate preparation (NOREL, BP70) was added to the D4 diet at 0.4%. At month 20, 6 fish per each dietary treatment were sampled for iTRAQ profiling and fingerprinting of intestinal mucus proteome. Mucus collected from anterior and posterior intestine segments was trypsin digested, labelled with iTRAQ reagents, isoelectrofocused and resolved by LC-MS/MS. More than 1000 proteins were unequivocally annotated and principal component analysis clearly separated anterior and posterior segments. The diet effect with changes in the abundance of approximately 120 proteins was restricted to anterior section with a reversion of the pattern of the extreme diet (D3 fish) with dietary butyrate supplementation. Butyrate supplementation also reversed the decrease of microbiotay diversity associated with D3 feeding, and led to a improvement the disease outcomes in fish challenged with Photobacterium damselae and the intestinal parasite Enteromyxum leei.
Project description:Four and a half LIM domains protein 2 (FHL2) is a co-regulator of gene transcription relevant for many signalling pathways and physiological processes. In this work, the role of FHL2 in cell differentiation has been investigated during in vitro mineralization using a Sparus aurata pre-osteoblast cell line (VSa16, [1]). A global microarray analysis was then performed to characterize the molecular pathways affected by FHL2 overexpression. [1] Pombinho, A.R., Laizé, V., Molha, D.M., Marques, S.M.P. and Cancela, M.L. (2004). Development of two bone-derived cell lines from the marine teleost Sparus aurata; evidence for extracellular matrix mineralization and cell-type specific expression of matrix Gla protein and osteocalcin. Cell Tissue Res. 315, 393-406.
Project description:Four and a half LIM domains protein 2 (FHL2) is a co-regulator of gene transcription relevant for many signalling pathways and physiological processes. In this work, the role of FHL2 in cell differentiation has been investigated during in vitro mineralization using a Sparus aurata pre-osteoblast cell line (VSa16, [1]). A global microarray analysis was then performed to characterize the molecular pathways affected by FHL2 overexpression. [1] Pombinho, A.R., LaizM-CM-), V., Molha, D.M., Marques, S.M.P. and Cancela, M.L. (2004). Development of two bone-derived cell lines from the marine teleost Sparus aurata; evidence for extracellular matrix mineralization and cell-type specific expression of matrix Gla protein and osteocalcin. Cell Tissue Res. 315, 393-406. Sparus aurata oligo-DNA microarray (GPL6467) was used to compare gene expression profiles between Control (WT) and FHL2-transfected (F2) VSa16 cell lines. Gene expression profiles of both WT and F2 cells were assessed either at T0 (initial confluent cultures) or T4 (4 weeks of cell confluency). Four (4) pools of cells were taken from both WT and F2 groups at each time point and stored in RNA later at -20M-BM-0C until RNA extraction.
Project description:Deciphering the dietary immunomodulatory effects of a medicinal plant leaf extract (MPLE) obateined from sage (Salvia officinalis, Lamiaceae) and lemon verbena (Lippia citriodora, Verbenaceae) upon the gut-associated lymphoid tissue (GALT) of gilthead seabream (Sparus aurata).
Project description:Agilent custom-made 8x60k 60-mer oligonucleotide microarray slides were used, covering unique sequences (21,093 annotated sequences and 4,299 unannotated sequences) of Sparus aurata transcriptomic data obtained from assembling of reads deposited in the NCBI SRA database corresponding to BioProject ID PRJNA391557, together with controls (8 transcripts of Sparus aurata plus Agilent controls). Two probes per sequence were used (in sense orientation for annotated sequences).
Project description:A gilthead sea bream (Sparus aurata) microarray platform was developed to identify brain gene expression profiles in response to environmental concentrations of human pharmaceuticals.
