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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datacite.alternateIdentifier.citationFRONTIERS IN VETERINARY SCIENCE,Vol.7,,2020
datacite.alternateIdentifier.doi10.3389/fvets.2020.586265
datacite.creatorValderrama, Ximena
datacite.creatorUlloa Leal, Cesar
datacite.creatorSilva Jiménez, Mauricio
datacite.creatorGoicochea, Jose
datacite.creatorApichela, Silvana
datacite.creatorArganaraz, Martin
datacite.creatorSari, Luciana
datacite.creatorPaiva, Luis
datacite.creatorRatto, Vicente Francisco
datacite.creatorRatto, Marcelo Hector
datacite.date2020
datacite.subject.englishMAPK
datacite.subject.englishU0126
datacite.subject.englishcorpus luteum
datacite.subject.englishfollicle
datacite.subject.englishSTAR
datacite.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.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.urihttp://repositoriodigital.uct.cl/handle/10925/3920
dc.language.isoen
dc.publisherFRONTIERS MEDIA SA
dc.sourceFRONTIERS IN VETERINARY SCIENCE
oaire.resourceTypeArticle
uct.catalogadorWOS
uct.indizacionSCI
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