Effect of anti-apoptotic drug Z-VAD-FMK on in vitro viability of dog follicles

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dc.contributor.authorCosta Pereira, Leda Maria [UNESP]
dc.contributor.authorThongkittidilok, Chommanart
dc.contributor.authorLopes, Maria Denise [UNESP]
dc.contributor.authorSongsasen, Nucharin
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionCenter for Species Survival
dc.date.accessioned2019-10-06T15:57:16Z
dc.date.available2019-10-06T15:57:16Z
dc.date.issued2018-12-01
dc.description.abstractIt is recognized that ovarian follicular atresia is associated with apoptosis, and the most important effector of cell death is caspase-3. The aim of this study was to investigate the influence of anti-apoptotic drug Z-VAD-FMK on in vitro follicle growth in the domestic dog. Ovaries were obtained from peri-pubertal and adult domestic dogs, and cortical fragments recovered and incubated on 1.5% (w/v) agarose gel blocks within a 24-well culture plate containing Minimum Essential Medium Eagle-Alpha Modification (αMEM) supplemented with 4.2 μg/mL insulin, 3.8 μg/mL transferrin, 5 ng/mL selenium, 2 mM L-glutamine, 100 μg/mL of penicillin G sodium, 100 μg/mL of streptomycin sulfate, 0.05 mM ascorbic acid, 10 ng/mL of FSH and 0.1% (w/v) polyvinyl alcohol in humidified atmosphere of 5% CO2 and 5% O2. The cortices were randomly allocated in six treatments: 1) 10 ng/mL EGF (EGF V0); 2) 10 ng/mL of EGF plus 1 mM Z-VAD-FMK (EGF V1); 3) 10 ng/mL of EGF and 10 mM Z-VAD-FMK (EGF V10); 4) 1 mM Z-VAD-FMK; 5) 10 mM Z-VAD-FMK and (6) no EGF and Z-VAD-FMK supplementation (Control). The cortices were processed for histology and assessed for viability (based on morphology), density of structurally normal follicles, and diameter immediately after collection (non-culture Control) or after 3 or 7 days of in vitro incubation. Evaluation of mRNA expression of Cas3 in fresh cortices and those incubated for 3 days was performed using real-time PCR. Histological analysis revealed that in vitro incubation decreased (P < 0.05) follicle viability and density compared to the fresh, non-culture control. Addition of 10 μM of Z-VAD-FMK alone to the culture medium sustained follicle viability at Day 3, but did not impact follicle diameter when compared to the other treatment groups (p < 0.001); however, the beneficial benefit of this anti-apoptotic drug diminished after 7 days of incubation. Furthermore, Z-VAD-FMK supplementation did not impact Cas3 expression. The findings demonstrated that dog ovarian tissues are highly susceptible to in vitro incubation and Z-VAD-FMK supported short-term survival of dog follicles enclosed within the ovarian cortex.en
dc.description.affiliationDepartment of Animal Reproduction and Veterinary Radiology Faculty of Veterinary Medicine FMVZ São Paulo State University - UNESP
dc.description.affiliationCenter for Species Survival, Smithsonian's National Zoological Park, Front Royal
dc.description.affiliationUnespDepartment of Animal Reproduction and Veterinary Radiology Faculty of Veterinary Medicine FMVZ São Paulo State University - UNESP
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipSmithsonian Institution
dc.description.sponsorshipIdFAPESP: 2013/21667-3
dc.description.sponsorshipIdFAPESP: 2014/19776-1
dc.description.sponsorshipIdFAPESP: 2015/17412-5
dc.format.extent124-129
dc.identifierhttp://dx.doi.org/10.1016/j.theriogenology.2018.09.012
dc.identifier.citationTheriogenology, v. 122, p. 124-129.
dc.identifier.doi10.1016/j.theriogenology.2018.09.012
dc.identifier.issn0093-691X
dc.identifier.lattes6666129914663018
dc.identifier.scopus2-s2.0-85053736377
dc.identifier.urihttp://hdl.handle.net/11449/188100
dc.language.isoeng
dc.relation.ispartofTheriogenology
dc.rights.accessRightsAcesso aberto
dc.sourceScopus
dc.subjectAnti-apoptotic drug
dc.subjectCaspase
dc.subjectDogs
dc.subjectFollicles
dc.titleEffect of anti-apoptotic drug Z-VAD-FMK on in vitro viability of dog folliclesen
dc.typeArtigo
unesp.author.lattes6666129914663018

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Artigos - Reprodução Animal e Radiologia Veterinária - FMVZ