| Texto completo | |
| Autor(es): |
Lima, Eliana R.
;
Sevilhano, Thais C. A.
;
Feitosa, Thais C.
;
Oliveira, Joao E.
;
Suzuki, Miriam F.
;
Torati, Lucas S.
;
Bartolini, Paolo
;
Peroni, Cibele N.
Número total de Autores: 8
|
| Tipo de documento: | Artigo Científico |
| Fonte: | FISHES; v. 10, n. 12, p. 12-pg., 2025-11-27. |
| Resumo | |
The cDNA sequences of the common gonadotrophic hormone alpha-subunit (ag-Gth alpha) and of the specific follicle-stimulating hormone beta-subunit (ag-Fsh beta) 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. (AU) | |
| Processo FAPESP: | 17/26369-1 - Síntese, purificação e caracterização das gonadotrofinas de pirarucu (Arapaima gigas): ag-FSH e ag-LH |
| Beneficiário: | João Ezequiel de Oliveira |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |
| Processo FAPESP: | 20/16549-5 - Estudos preliminares relativos à aplicação da terapia gênica de hormônio de crescimento em Arapaima gigas |
| Beneficiário: | Paolo Bartolini |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |