Title:	Extensive characterization of Arapaima gigas follicle-stimulating hormone (ag-Fsh) synthesized in HEK293 cells.
Authors:	LIMA, E. R. SEVILHANO, T. C. A. FEITOSA, T. C. OLIVEIRA, J. E. SUZUKI, M. F. TORATI, L. S. BARTOLINI, P. PERONI, C. N.
Affiliation:	ELIANA R. LIMA, UNIVERSIDADE DE SÃO PAULO; THAIS C. A. SEVILHANO, UNIVERSIDADE DE SÃO PAULO; THAIS C. FEITOSA, UNIVERSIDADE DE SÃO PAULO; JOÃO E. OLIVEIRA, UNIVERSIDADE DE SÃO PAULO; MIRIAM F. SUZUKI, UNIVERSIDADE DE SÃO PAULO; LUCAS SIMON TORATI, CNPASA; PAOLO BARTOLINI, UNIVERSIDADE DE SÃO PAULO; CIBELE N. PERONI, UNIVERSIDADE DE SÃO PAULO.
Date Issued:	2025
Citation:	Fishes, v. 10, n. 12, p. 607, 2025.
Description:	The cDNA sequences of the common gonadotrophic hormone α-subunit (ag-Gthα) and of the specific follicle-stimulating hormone β-subunit (ag-Fshβ) of the giant Amazonian fish Arapaima gigas have been previously isolated by our research group. A synthesis of ag-Fsh in HEK293 cells and its purification and preliminary characterization were also carried out. In the present work, ag-Fsh was obtained and purified from the same host cells, and an extensive physical chemical characterization was performed via RP-HPLC, HPSEC, and MALDI-TOF-MS. Ag-Fsh, when compared to human FSH (hFSH), showed a higher hydrophobicity by RP-HPLC and a higher molecular mass (MM) via HPSEC. The same higher MM was also confirmed via MALDI-TOF-MS: 35,353 Da for ag-FSH against 31,969 Da for hFSH. Its immunological activity was also confirmed via an hFSH ELISA, in comparison with the highly purified pituitary preparation of hFSH-AFP7298A, from the National Hormone and Pituitary Program (NHPP-USA), with a clear response that was, however, 1560-fold lower when compared to the purest preparation. Finally, an in vitro bioassay, based on the stimulated release of 11-ketotestosterone (11-KT) from immature A. gigas testis, quantified ag-Fsh biological activity in comparison with human chorionic gonadotropin (hCG) and with human pituitary FSH-AFP7298A, showing a potency clearly higher than that of hCG. This suggests that injections of ag-Fsh in A. gigas and ag-Fsh cDNA gene therapy applications could be used for improving the reproductive functions of this threatened species.
Thesagro:	Hormônio
NAL Thesaurus:	Arapaima gigas
Keywords:	Cetotestosterona
DOI:	https://doi.org/10.3390/fishes10120607
Type of Material:	Artigo de periódico
Access:	openAccess
Appears in Collections:	Artigo em periódico indexado (CNPASA)