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Synthesis, purification and characterization of the gonadotropins of pirarucu (Arapaima gigas): ag-FSH and ag-LH

Abstract

Arapaima gigas lives in the Amazon, São Francisco and North Eastern basins of Brazil and is an important species for the alimentation and commercial breeding of the region. It is considered one of overexploited fishes by the Brazilian Institute of Environment (IBAMA), in danger of extinction and in the red list as "Data deficient". We must consider, moreover, that fishes grown in captivity frequently present difficulty in ovulation and spermatogenesis and fertility problems that are related to the hormones of the hypothalamus-hypophysis axis. With the aim of investigating A. gigas physiology, controlling and improving its reproduction especially in captivity, the goal of the present project is to synthesize, purify and extensively characterize the gonadotrophic hormones of this species: ag-FSH and ag-LH.Our research group, having already cloned and characterized the cDNA of the common ±-subunit, with consequent publication of the data obtained in the journal Fish Physiology and Biochemistry, carried out also a FAPESP Project (Proc. N. 2012/10779-2) whose goal, besides the cloning of the two cDNAs of ag-FSH-² and ag-LH-² chains, was also the expression and characterization of the two hormones. This synthesis, unfortunately, was not attained, as described in the last progress report, but its preliminary expression in HEK293 cells and characterization was then obtained, only for ag-FSH, after the conclusion of the Project as reported in a recent publication in PLOS ONE. The goal of the present Project is, therefore, the synthesis, purification and characterization of the two hormones, including all physical chemical and biological testing that are necessary for their utilization in the areas of endocrinology and reproduction of the same A. gigas, the biggest sweet water fish of the world, which is very important for food and economy of the region. The possibility of the application of these two hormones in Arapaima gigas is already under study in collaboration with the team of "Embrapa Pesca e Aquicultura" of Palmas (Tocantins, Brazil) that is specialized in the study of the reproduction conditions of this fish.A Ph.D. fellowship request (just for the last two years) was also sent to FAPESP for the M.S. Thais Sevilhano, who will be involved with most of these activities. (AU)

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