Bottom-up regulation of phytoplankton in the Guadiana estuary

Tese dout., Ciências do Mar (Ecologia Marinha), Faculdade de Ciências e Tecnologia, Univ. do Algarve, 2010

Effects of nutrient and light enrichment on phytoplankton growth

Alterations of freshwater flow regimes and increasing eutrophication lead to alterations in light availability and nutrient loading into adjacent estuaries and coastal areas. Phytoplankton community respond to these changes in many ways. Harmful phytoplankton blooms, for instance, may be a consequence of changes in nutrient supply, as well as the replacement of some phytoplankton species (like diatoms, that contribute for the development of large fish and shellfish populations) by ohers (like cyanobacteria, that may be toxic and represent an undesirable food source for higher trophic levels). Nutrient and light enrichment experiments allow us to understand and predict the effects of eutrophication on the growth of phytoplankton. This is a fundamental tool in water management issues, since it enables the prediction of changes in the phytoplankton community that may be harmful to the whole ecosystem, and the design of mitigation strategies (Zalewski 2000).

Identification of novel molecular determinants of tissue mineralization in fish

The identification of genes involved in signaling and regulatory pathways, and matrix formation is paramount to the better understanding of the complex mechanisms of bone formation and mineralization, and critical to the successful development of therapies for human skeletal disorders. To achieve this objective, in vitro cell systems derived from skeletal tissues and able to mineralize their extracellular matrix have been used to identify genes differentially expressed during mineralization and possibly new markers of bone and cartilage homeostasis. Using cell systems of fish origin and techniques such as suppression subtractive hybridization and microarray hybridization, three genes never associated with mechanisms of calcification were identified: the calcium binding protein S100-like, the short-chain dehydrogenase/reductase sdr-like and the betaine homocysteine S-methyltransferase bhmt3. Analysis of the spatial-temporal expression of these 3 genes by qPCR and in situ hybridization revealed: (1) the up-regulation of sdr-like transcript during in vitro mineralization of gilthead seabream cell lines and its specificity for calcified tissues and differentiating osteoblasts; (2) the up-regulation of S100-like and the down-regulation of bhmt3 during in vitro mineralization and the central role of both genes in cartilaginous tissues undergoing endo/perichondral mineralization in juvenile fish. While expression of S100-like and bhmt3 was restricted to calcified tissues, sdr-like transcript was also detected in soft tissues, in particular in tissues of the gastrointestinal tract. Functional analysis of gene promoters revealed the transcriptional regulation of the 3 genes by known regulators of osteoblast and chondrocyte differentiation/mineralization: RUNX2 and RAR (sdr-like), ETS1 (s100-like; bhmt3), SP1 and MEF2c (bhmt3). The evolutionary relationship of the different orthologs and paralogs identified within the scope of this work was also inferred from taxonomic and phylogenetic analyses and revealed novel protein subfamilies (S100-like and Sdr-like) and the explosive diversity of Bhmt family in particular fish groups (Neoteleostei). Altogether our results contribute with new data on SDR, S100 and BHMT proteins, evidencing for the first time the role for these three proteins in mechanisms of mineralization in fish and emphasized their potential as markers of mineralizing cartilage and bone in developing fish.

Identification of novel molecular determinants of tissue mineralization in fish

Tese de doutoramento, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2015