β-NGF Stimulates Steroidogenic Enzyme and VEGFA Gene Expression, and Progesterone Secretion via ERK 1/2 Pathway in Primary Culture of Llama Granulosa Cells.

Autor:	Valderrama X; Instituto de Investigaciones Agropecuarias (INIA), Osorno, Chile., Ulloa-Leal C; Institute of Animal Science, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia, Chile., Silva ME; Department of Veterinary Sciences and Public Health, Faculty of Natural Resources, Universidad Catolica de Temuco, Temuco, Chile., Goicochea J; Department of Surgery and Reproductive Biotechnology, Faculty of Veterinary Medicine and Zootechnics, Universidad Nacional Hermilio Valdizán, Huánuco, Peru., Apichela S; Instituto Superior de Investigaciones Biológicas (INSIBIO), CONICET-UNT, Facultad de Bioquímica, Instituto de Biología 'Dr. Francisco D. Barbieri,' Química y Farmacia, UNT, San Miguel de Tucumán, Argentina., Argañaraz M; Instituto Superior de Investigaciones Biológicas (INSIBIO), CONICET-UNT, Facultad de Bioquímica, Instituto de Biología 'Dr. Francisco D. Barbieri,' Química y Farmacia, UNT, San Miguel de Tucumán, Argentina., Sari L; Instituto Superior de Investigaciones Biológicas (INSIBIO), CONICET-UNT, Facultad de Bioquímica, Instituto de Biología 'Dr. Francisco D. Barbieri,' Química y Farmacia, UNT, San Miguel de Tucumán, Argentina., Paiva L; Institute of Animal Science, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia, Chile., Ratto VF; Institute of Animal Science, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia, Chile., Ratto MH; Institute of Animal Science, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia, Chile.
Jazyk:	angličtina
Zdroj:	Frontiers in veterinary science [Front Vet Sci] 2020 Oct 23; Vol. 7, pp. 586265. Date of Electronic Publication: 2020 Oct 23 (Print Publication: 2020).
DOI:	10.3389/fvets.2020.586265
Abstrakt:	The beta-nerve growth factor (β-NGF) from llama seminal plasma exerts ovulatory and luteotrophic effects following intramuscular or intrauterine infusion in llamas and alpacas. In this study, we investigate the in vitro effect of llama β-NGF on the expression of genes involved in angiogenesis and progesterone synthesis as well as progesterone release in preovulatory llama granulosa cells; we also determine whether these changes are mediated via the ERK1/2 signaling pathway. From adult female llamas, we collected granulosa cells from preovulatory follicles by transvaginal ultrasound-guided follicle aspiration; these cells were pooled and incubated. After 80% confluence, the cultured granulosa cells were treated with β-NGF, β-NGF plus the MAPK inhibitor U0126, or luteinizing hormone, and the abundance of angiogenic and steroidogenic enzyme mRNA transcripts were quantified after 10 and 20 h by RT-qPCR. We also quantified the progesterone concentration in the media after 48 h by radioimmunoassay. We found that application of β-NGF increases the abundance of mRNA transcripts of the vascular endothelial growth factor ( VEGFA ) and the steroidogenic enzymes cytochrome P450 side-chain cleavage (P450scc/ CYP11A1 ), steroidogenic acute regulatory protein ( STAR ), and 3β-hydroxysteroid dehydrogenase ( HSD3B1 ) at 10 and 20 h of treatment. Application of the MAPK inhibitor U0126 resulted in downregulation of the genes encoding these enzymes. β-NGF also enhanced progesterone synthesis, which was prevented by the prior application of the MAPK inhibitor U0126. Finally, western blot analysis confirmed that β-NGF activates the ERK1/2 signaling pathway. In conclusion, our results indicate that β-NGF exerts direct luteotropic effects on llama ovarian tissue via the ERK 1/2 pathway. (Copyright © 2020 Valderrama, Ulloa-Leal, Silva, Goicochea, Apichela, Argañaraz, Sari, Paiva, Ratto and Ratto.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu