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

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dc.contributor.authorValderrama, Ximena
dc.contributor.authorUlloa Leal, Cesar
dc.contributor.authorSilva Jiménez, Mauricio
dc.contributor.authorGoicochea, Jose
dc.contributor.authorApichela, Silvana
dc.contributor.authorArganaraz, Martin
dc.contributor.authorSari, Luciana
dc.contributor.authorPaiva, Luis
dc.contributor.authorRatto, Vicente Francisco
dc.contributor.authorRatto, Marcelo Hector
dc.date2020
dc.date.accessioned2021-04-30T17:05:04Z
dc.date.available2021-04-30T17:05:04Z
dc.description.abstractThe beta-nerve growth factor (beta-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 beta-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 beta-NGF, beta-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 beta-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 3 beta-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. beta-NGF also enhanced progesterone synthesis, which was prevented by the prior application of the MAPK inhibitor U0126. Finally, western blot analysis confirmed that beta-NGF activates the ERK1/2 signaling pathway. In conclusion, our results indicate that beta-NGF exerts direct luteotropic effects on llama ovarian tissue via the ERK 1/2 pathway.
dc.identifier.citationFRONTIERS IN VETERINARY SCIENCE,Vol.7,,2020
dc.identifier.doi10.3389/fvets.2020.586265
dc.identifier.urihttp://repositoriodigital.uct.cl/handle/10925/3920
dc.language.isoen
dc.publisherFRONTIERS MEDIA SA
dc.sourceFRONTIERS IN VETERINARY SCIENCE
dc.subject.englishMAPK
dc.subject.englishU0126
dc.subject.englishcorpus luteum
dc.subject.englishfollicle
dc.subject.englishSTAR
dc.titlebeta-NGF Stimulates Steroidogenic Enzyme and VEGFA Gene Expression, and Progesterone Secretion via ERK 1/2 Pathway in Primary Culture of Llama Granulosa Cells
dc.typeArticle
uct.catalogadorWOS
uct.indizacionSCI
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