Cloning, expression and characterization of Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) of pirarucu (Arapaima gigas)

• Grant number: 12/10779-2
• Support Opportunities: Regular Research Grants
• Start date: May 01, 2013
• End date: April 30, 2016
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Paolo Bartolini
• Grantee: Paolo Bartolini
• Host Institution: Instituto de Pesquisas Energéticas e Nucleares (IPEN). Secretaria de Desenvolvimento Econômico (São Paulo - Estado). São Paulo , SP, Brazil
• Associated researchers: Carlos Roberto Jorge Soares ; Cibele Nunes Peroni ; João Ezequiel de Oliveira ; Kayo Okazaki ; Maria Teresa de Carvalho Pinto Ribela ; Miriam Fussae Suzuki

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, being considered one of overexploited fishes by the Brazilian Institute of Environment (IBAMA) and being also included in the list of endangered fishes according to the Convention on International Trade of Endangered Species of Wild Fauna and Flora (CITES, Appendix II). We must consider, moreover, that fishes grown in captivity frequently present difficulty in ovulation and spermatogenesis. These fertility problems are related to the hormones of the hypothalamus-hypophysis axis that include the two gonadotropins (GTH):follicle-stimulating (FSH) and luteinizing (LH) hormone. We emphasize that different species of fishes already had their FSH and LH cloned and sequenced, no one, however, belonging to the order of osteoglossiformes or even to the superorder of osteoglossomorpha, to which the genus Arapaima belongs. With the goal of investigating the physiology of reproduction of A. gigas, being thus able to control and improve its reproduction, we decided in the present Project to clone and express FSH and LH of this species. Considering that the sequence of the alpha subunit of A. gigas gonadotropins was already obtained in our laboratory, the cDNAs coding for the two beta subunits (FSH and LH) of these hormones will be now isolated and sequenced, for the first time from A. gigas pituitaries, being then inserted into vectors that are adequate for their expression in Chinese hamster ovary (CHO) cells. This way, we will synthesize, purify and characterize recombinant FSH and LH (r-agFSH and r-agLH)of A. gigas, the largest freshwater fish of the world, which is very important for food and economy of the region. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

SEVILHANO, THAIS; DE CARVALHO, ROBERTO FEITOSA; DE JESUS OLIVEIRA, NELIO ALESSANDRO; OLIVEIRA, JOAO EZEQUIEL; MALTAROLLO, VINICIUS GONCALVES; TROSSINI, GUSTAVO; GARCEZ, RIVIANE; BARTOLINI, PAOLO. Molecular cloning and characterization of pirarucu (Arapaima gigas) follicle-stimulating hormone and luteinizing hormone beta-subunit cDNAs. PLoS One, v. 12, n. 8, AUG 28 2017. (12/10779-2)

RIBELA, MARIA TERESA C. P.; DAMIANI, RENATA; SILVA, FELIPE D.; LIMA, ELIANA R.; OLIVEIRA, JOAO E.; PERONI, CIBELE N.; TORJESEN, PETER A.; SOARES, CARLOS R.; BARTOLINI, PAOLO. N-Glycoprofiling Analysis for Carbohydrate Composition and Site-Occupancy Determination in a Poly-Glycosylated Protein: Human Thyrotropin of Different Origins. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 18, n. 2, FEB 2017. (15/26058-0, 11/07289-0, 12/10779-2)