title: Effects of frutalin on early follicle morphology, ultrastructure and gene expression in cultured goat ovarian cortical tissue.
abstract: Frutalin is a galactose-binding lectin that has an irreversible cytotoxic effect on HeLa cervical cancer cells, by inducing apoptosis and inhibiting cell proliferation. It was previously shown that after in vitro incubation, frutalin is internalized into HeLa cells nucleus, which indicates that frutalin apoptosis-inducing activity might be linked with its nuclear localization. Considering that drugs commonly used for cancer treatment have a deleterious effect on germ cells, the aim of this study was to evaluate the effect of frutalin on the activation, survival, ultrastructure and gene expression in follicles cultured within ovarian tissue. Goat ovarian fragments were cultured for 6 days in α-MEM⁺ alone or supplemented with frutalin (1, 10, 50, 100 or 200 µg/ml). Non-culturad and cultured tissues were processed for histological and ultrastructural analysis and they were also stored to evaluate the expression of anti- and pro-apoptotic genes by quantitative polymerase chain reaction (qPCR). The results showed that the frutalin, at all concentrations tested, reduced follicular survival when compared with control medium. Higher concentrations of frutalin (50, 100 or 200 µg/ml) also reduced follicular survival when compared with those tissues cultured with 1 or 10 µg/ml of frutalin. The ultrastructural analysis showed that atretic cultured follicles had retracted oocytes and a large number of vacuoles spread throughout the cytoplasm. In addition, signs of damage of mitochondrial membranes and cristae were observed. Moreover, although a dose-response effect on gene expression has not been observed, when compared with tissues culture in control medium, the presence of frutalin increased in mRNA expression pro-apoptotic genes. In conclusion, frutalin reduces follicular survival at all concentrations tested, its effects being more pronounced when high concentrations of this lectin (50, 100 and 200 µg/ml) are used. Gene expression profile and ultrastrutural features of cultured follicles suggest that follicular death in goat ovarian tissue cultured in presence of frutalin occurs via necrosis.
authors.affiliation: Biotechnology Nucleus of Sobral (NUBIS), Federal University of Ceará, Sobral, Ceará, Brazil.
authors.forename: Maria A A
authors.initials: MAA
authors.lastname: Soares
authors.affiliation: Biotechnology Nucleus of Sobral (NUBIS), Federal University of Ceará, Sobral, Ceará, Brazil.
authors.forename: José J N
authors.initials: JJN
authors.lastname: Costa
authors.affiliation: Reproductive Physiology Laboratory, Department of Animal Science, Federal University of Lavras, Lavras, Minas Gerais, Brazil.
authors.forename: Gisvani L
authors.initials: GL
authors.lastname: Vasconcelos
authors.affiliation: Biotechnology Nucleus of Sobral (NUBIS), Federal University of Ceará, Sobral, Ceará, Brazil.
authors.forename: Regislane P
authors.initials: RP
authors.lastname: Ribeiro
authors.affiliation: Reproductive Physiology Laboratory, Department of Animal Science, Federal University of Lavras, Lavras, Minas Gerais, Brazil.
authors.forename: José C
authors.initials: JC
authors.lastname: Souza
authors.affiliation: Molecular Biotechnology Laboratory (LabBMol), Federal University of Ceará, Fortaleza, Ceará, Brazil.
authors.forename: André L C
authors.initials: ALC
authors.lastname: Silva
authors.affiliation: Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Robert
authors.initials: R
authors.lastname: Van den Hurk
authors.affiliation: Biotechnology Nucleus of Sobral (NUBIS), Federal University of Ceará, Sobral, Ceará, Brazil. jrvsilva@ufc.br, roberto_viana@yahoo.com.
authors.forename: José R V
authors.initials: JRV
authors.lastname: Silva
mesh_terms: D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000633:toxicity*; D016923:Cell Death / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005260:Female; D037161:Galectins / Q000633:toxicity*; D005786:Gene Expression Regulation; D006041:Goats; D009336:Necrosis; D006080:Ovarian Follicle / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D010053:Ovary / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D013997:Time Factors; D046509:Tissue Culture Techniques
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D037161:Galectins; C570811:frutalin