Valentina Chini

Genes expressed in Blue Fin Tuna (Thunnus thynnus) liver and gonads.

Blue Fin Tuna (BFT), Thunnus thynnus, has been seriously endangered by global massive overfishing and by the pollution of marine environment. Feeding and fattening of caught tuna in marine cages is a recent resource, but the development of a self-sustained aquaculture activity, being independent from the supply of wild fish, is required from both industrial and conservation perspectives. At this scope, several technical problems have to be solved and the control of reproduction is the cardinal one. Beside the technological developments of farming facilities and protocols, a molecular approach seems promising for the studies of appropriate nutritional strategies, reproduction physiology and animal welfare, as well as lifestyle and response to endocrine disruptor pollutants. In this context, we have started an EST project on this species sequencing 2743, 2907, and 3014 clones from expression libraries of ovary, testis and liver, respectively, and 1499 clones from an ovary normalized library. Thanks to this project, we have identified several sequences with known function in other organisms, but not previously described in this species. Among the new genes, 712 were found only in the expression library of the ovary, 613 in that of the testis and 318 in that of the liver, while 324 additional genes were shared by two or more expression libraries; other 127 genes not found in the expression libraries were obtained from the ovary normalized library. This represents a contribution to the knowledge of the molecular basis of BFT and a necessary step for facilitating further molecular studies on this species. Accession numbers: EC 091633 to EC 093160; EG 629962 to EG 631176; EC 917676 to EC 919417; EG 999340 to EG 999999; EH 000001 to EH 000505; EH 667253 to EH 668984; EL 610526 to EL 611807; EC 42144 to EC 422414; and EH 379568 to EH 380065.

Evaluation of SAT-1, SAT-2 and GalNAcT-1 mRNA in colon cancer by real-time PCR

By qualitative and quantitative PCR, we evaluated the expression of three messengers coding for SAT-1, SAT-2 and GalNAcT-1 in human samples of intestinal cancer and some cell lines (breast cancer and melanomas). Qualitative PCR demonstrated, in human tissues but not in the cell lines examined, the presence of an mRNA that lacks hexon 3; experiments performed on transfected SKMEL-28 excluded a regulative role of this noncanonical mRNA. Data from real-time PCR, statistically analysed by ANOVA indicated that the mRNA expression of all the considered glycosyltransferases (SAT-1, SAT-2 and GalNAcT-1) was significantly different in tumours versus their own control. The ganglioside patterns in the examined samples did not correlate with mRNA expression; this finding demonstrates that ganglioside expression is the result of a very complex balance between anabolic and catabolic enzyme activities.Although this study is still preliminary, it opens a new possibility for neoplastic prognosis finding potential molecular markers among the mRNAs that codify for glycosyltransferases.

EST projects in aquaculture: sea bass, red tuna and perch

Riassunto Progetto EST in acquacoltura: spigola, tonno rosso e persico. Queste tre specie stanno richiamando un sempre maggior interesse. Si è reso quindi necessario effettuare un programma che permetta di valutare le abitudini comportamentali dei pesci di allevamento e gli eventuali problemi nelle varie fasi del ciclo biologico indagando, per esempio, l’espressione di geni coinvolti nella maturazione dei gameti e nella fecondazione e/o quelli che hanno un ruolo nella risposta di tipo biologico agli stressori ambientali. A tal fine abbiamo dato il via a questo progetto preparando una library di cDNA di fegato di branzino e sequenziando alcune centinaia di cloni il 55% dei quali risultano essere sequenze singole, mentre i restanti vanno a formare dei contig. Contemporaneamente è in corso uno studio sulle gonadi di tonno rosso e sul fegato di persico. L’ obiettivo di questo progetto è quello di creare database di sequenze di cDNA di queste tre specie di pesci ad interesse commerciale. Di tali library verranno sequenziati circa 5.000 cloni; le sequenze ottenute, depositate in banca dati, saranno disponibili via Internet a tutta la comunità scientifica che le potrà utilizzare per le ricerche più disparate.