Página 1 dos resultados de 8807 itens digitais encontrados em 0.012 segundos

Sequential analysis of global gene expression profiles in immature and in vitro matured bovine oocytes: potential molecular markers of oocyte maturation

MAMO, Solomon; CARTER, Fiona; LONERGAN, Patrick; LEAL, Claudia L. V.; NAIB, Abdullah Al; MCGETTIGAN, Paul; MEHTA, Jai P.; EVANS, Alexander C. O.; FAIR, Trudee
Fonte: BIOMED CENTRAL LTD Publicador: BIOMED CENTRAL LTD
Tipo: Artigo de Revista Científica
ENG
Relevância na Pesquisa
36.89%
Background: Without intensive selection, the majority of bovine oocytes submitted to in vitro embryo production (IVP) fail to develop to the blastocyst stage. This is attributed partly to their maturation status and competences. Using the Affymetrix GeneChip Bovine Genome Array, global mRNA expression analysis of immature (GV) and in vitro matured (IVM) bovine oocytes was carried out to characterize the transcriptome of bovine oocytes and then use a variety of approaches to determine whether the observed transcriptional changes during IVM was real or an artifact of the techniques used during analysis. Results: 8489 transcripts were detected across the two oocyte groups, of which similar to 25.0% (2117 transcripts) were differentially expressed (p < 0.001); corresponding to 589 over-expressed and 1528 under-expressed transcripts in the IVM oocytes compared to their immature counterparts. Over expression of transcripts by IVM oocytes is particularly interesting, therefore, a variety of approaches were employed to determine whether the observed transcriptional changes during IVM were real or an artifact of the techniques used during analysis, including the analysis of transcript abundance in oocytes in vitro matured in the presence of a-amanitin. Subsets of the differentially expressed genes were also validated by quantitative real-time PCR (qPCR) and the gene expression data was classified according to gene ontology and pathway enrichment. Numerous cell cycle linked (CDC2...

Effects of gonadotropin-exposed medium with high concentrations of progesterone and estradiol-17 beta on in vitro maturation of canine oocytes

VANNUCCHI, Camila Infantosi; FAUSTINO, Marcelo; MARQUES, Mariana Groke; NICHI, Marcilio; ASSUMPCAO, Mayra Elena Ortiz D`Avila; VISINTIN, Jose Antonio
Fonte: SPRINGER Publicador: SPRINGER
Tipo: Artigo de Revista Científica
ENG
Relevância na Pesquisa
36.91%
During the process of maturation in the oviduct, canine oocytes in the germinal vesicle stage are exposed to decreasing levels of estradiol-17 beta and increasing levels of progesterone. However, hormone concentrations in the microenvironments in which they act are higher than serum concentrations. Therefore, the aim of the present study was to compare the meiotic competence of canine oocytes harvested from anestrous bitches in culture medium containing high concentrations (20 mu g ml(-1)) of estradiol-17 beta and/or progesterone in association to gonadotropins (luteinizing hormone and follicle-stimulating hormone) using three different maturation periods (48, 72, and 96 h). Oocytes were cultured in tissue culture medium (TCM-199) and arranged in four experimental groups: group control, group E2 (estradiol-17 beta), group P4 (progesterone), and group E2 + P4. Regardless of the maturation period, groups P4 and E2 + P4 presented statistically higher rate of germinal vesicle breakdown oocytes compared to the group control and group E2. There were no significant differences among groups on germinal vesicle, metaphase I, metaphase II, and degenerated or unidentifiable oocytes rates. The mean percentage of metaphase II oocytes was higher at 96 h when compared to 72 h. Results of the present research indicate no influence of estradiol-17 beta supplementation...

Messenger RNA expression of Pabpnl1 and Mbd3l2 genes in oocytes and cleavage embryos

BIASE, Fernando Henrique; MARTELLI, Lucia; PUGA, Renato; GIULIATTI, Silvana; SANTOS-BIASE, Weruska Karyna Freitas; MERIGHE, Giovana Krempel Fonseca; MEIRELLES, Flavio Vieira
Fonte: ELSEVIER SCIENCE INC Publicador: ELSEVIER SCIENCE INC
Tipo: Artigo de Revista Científica
ENG
Relevância na Pesquisa
36.93%
Objective: To identify genes specifically expressed in mammalian oocytes using an in silico subtraction, and to characterize the mRNA patterns of selected genes in oocytes, embryos, and adult tissues. Design: Comparison between oocyte groups and between early embryo stages. Setting: Laboratories of embryo manipulation and molecular biology from Departamento de Genetica (FMRP) and Departamento de Ciencias Basicas (FZEA) - University of Sao Paulo. Sample(s): Oocytes were collected from slaughtered cows for measurements, in vitro fertilization, and in vitro embryo culture. Somatic tissue, excluding gonad and uterus tissue, was collected from male and female cattle. Main Outcome Measure(s): Messenger RNA levels of poly(A)-binding protein nuclear-like 1 (Pabpnl1) and methyl-CpG-binding domain protein 3-like 2 (Mbd3l2). Result(s): Pabpnl1 mRNA was found to be expressed in oocytes, and Mbd3l2 transcripts were present in embryos. Quantification of Pabpnl1 transcripts showed no difference in levels between good-and bad-quality oocytes before in vitro maturation (IVM) or between good-quality oocytes before and after IVM. However, Pabpnl1 transcripts were not detected in bad-quality oocytes after IVM. Transcripts of the Mbd3l2 gene were found in 4-cell...

Endothelial and inducible nitric oxide synthases in oocytes of cattle

PIRES, P. R. L.; SANTOS, N. P.; ADONA, P. R.; NATORI, M. M.; SCHWARZ, K. R. L.; BEM, T. H. C. de; LEAL, C. L. V.
Fonte: ELSEVIER SCIENCE BV Publicador: ELSEVIER SCIENCE BV
Tipo: Artigo de Revista Científica
ENG
Relevância na Pesquisa
36.95%
Nitric oxide (NO) is a chemical messenger generated by the activity of the nitric oxide synthases (NOS). The NOS/NO system appears to be involved in oocyte maturation, but there are few studies on gene expression and protein activity in oocytes of cattle. The present study aimed to investigate gene expression and protein activity of NOS in immature and in vitro matured oocytes of cattle. The influence of pre-maturation culture with butyrolactone I in NOS gene expression was also assessed. The following experiments were performed: (1) detection of the endothelial (eNOS) and inducible (iNOS) isoforms in the ovary by immunohistochemistry; (2) detection of eNOS and iNOS in the oocytes before and after in vitro maturation (W) by immunofluorescence; (3) eNOS and iNOS mRNA and protein in immature and in vitro matured oocytes, with or without pre-maturation, by real time PCR and Western blotting, respectively; and (4) NOS activity in immature and in vitro matured oocytes by NADPH-diaphorase. eNOS and iNOS were detected in oocytes within all follicle categories (primary, secondary and tertiary), and other compartments of the ovary and in the cytoplasm of immature and in vitro matured oocytes. Amount of mRNA for both isoforms decreased after IVM but was maintained after pre-maturation culture. The NOS protein was detected in immature (pre-mature or not) and was still detected in similar amount after pre-maturation and maturation for both isoforms. NOS activity was detected only in part of the immature oocytes. In conclusion...

Gene Expression in Bovine Oocytes and Cumulus Cells After Meiotic Inhibition with the Cyclin-Dependent Kinase Inhibitor Butyrolactone I

Leal, C. L. V.; Mamo, S.; Fair, T.; Lonergan, P.
Fonte: WILEY-BLACKWELL; HOBOKEN Publicador: WILEY-BLACKWELL; HOBOKEN
Tipo: Artigo de Revista Científica
ENG
Relevância na Pesquisa
36.95%
Contents The aim of this study was to determine the effect of temporary inhibition of meiosis using the cyclin-dependent kinase inhibitor butyrolactone I (BLI) on gene expression in bovine oocytes and cumulus cells. Immature bovine cumulusoocyte complexes (COCs) were assigned to groups: (i) Control COCs collected immediately after recovery from the ovary or (ii) after in vitro maturation (IVM) for 24 h, (iii) Inhibited COCs collected 24 h after incubation with 100 mu m BLI or (iv) after meiotic inhibition for 24 h followed by IVM for a further 22 h. For mRNA relative abundance analysis, pools of 10 denuded oocytes and respective cumulus cells were collected. Transcripts related to cell cycle regulation and oocyte competence were evaluated in oocytes and cumulus cells by quantitative real-time PCR (qPCR). Most of the examined transcripts were downregulated (p < 0.05) after IVM in control and inhibited oocytes (19 of 35). Nine transcripts remained stable (p > 0.05) after IVM in control oocytes; only INHBA did not show this pattern in inhibited oocytes. Seven genes were upregulated after IVM in control oocytes (p < 0.05), and only PLAT, RBP1 and INHBB were not upregulated in inhibited oocytes after IVM. In cumulus cells, six genes were upregulated (p < 0.05) after IVM and eight were downregulated (p < 0.05). Cells from inhibited oocytes showed the same pattern of expression regarding maturation profile...

Ativação partenogenética de oócitos bovinos jovens com ionomicina e 6-dimetilaminopurina associado ou não ao estrôncio; Parthenogenetic activation of young bovine oocytes with ionomycin and 6-dimethylaminopurine associated or not with strontium

Porciuncula, Patrícia Marafon
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: Dissertação de Mestrado Formato: application/pdf
Publicado em 05/10/2007 PT
Relevância na Pesquisa
36.89%
Tendo em vista a possibilidade da associação de agentes ativadores aliada à influência da idade do oócito no desenvolvimento embrionário, a proposta deste trabalho foi estudar a relação entre o envelhecimento do oócito e a ativação partenogenética. Para tal, oócitos bovinos foram maturados in vitro por um período de 22 h (jovens) e 28 h (envelhecidos). Em seguida, os dois grupos de oócitos foram ativados com os tratamentos: ID3: ionomicina (5 µM por 5 min) + 6DMAP (2 mM por 3 h de incubação); ID6: ionomicina (5 µM por 5 min) + 6DMAP (2 mM por 6 h de incubação); IDS: ionomicina + associação 6DMAP (2 mM) e estrôncio (20 mM; SrCl2) por 6 h e IDSS: ionomicina + (6DMAP+Sr) nas primeiras 3 h, seguido de lavagem e incubação com estrôncio (Sr2+) isoladamente por mais 3 h de incubação. O grupo controle foi cultivado na ausência de qualquer agente ativador (ativação espontânea). Após a ativação os oócitos foram avaliados quanto a: 1) taxa de ativação (formação de pronúcleo às 12 hpa); 2) atividade do fator promotor de maturação (MPF) e proteína cinase ativada por mitógeno (MAPK) nos intervalos de 5 min, 3 h, 6 h e 10 h; 3) desenvolvimento embrionário (taxa de clivagem às 48 h, dia 7 e 9 taxas de blastocisto e eclosão); 4) qualidade dos embriões (dia 9 pelo número total de células...

Effect of capacitation of stallion sperm with polyvinylalcohol or bovine serum albumin on penetration of bovine zona-free or partially zona-removed equine oocytes

Choi, Y. H.; Landim-Alvarenga, F. C.; Seidel, G. E.; Squires, E. L.
Fonte: Universidade Estadual Paulista Publicador: Universidade Estadual Paulista
Tipo: Artigo de Revista Científica Formato: 2080-2087
ENG
Relevância na Pesquisa
36.97%
Experiments were conducted to study effects of macromolecules on stallion sperm capacitation and fertilization as determined by penetration of bovine zona-free and equine partially zona-removed oocytes. Stallion sperm were capacitated in TYH medium (modified Krebs-Ringer bicarbonate) supplemented with either 1 mg/mL of polyvinylalcohol (PVA) or 4 mg/ mL of BSA. Capacitation was induced with 8 bromoadenosine cyclic monophosphate (8BrcAMP; 0.5 mM) alone or in combination with 0.1 μM of ionomycin. Intraspecies gametes were co-incubated in TYH/PVA or TYH/ BSA for 18 to 20 h. For zona-free bovine oocytes, penetration rate (35%) with the combination of 8BrcAMP and ionomycin in PVA-containing medium was higher (P < 0.05) than any treatment in BSA-containing medium (5 to 6%). A similar study was conducted using equine oocytes with partially removed zonae. Sperm capacitated and used for in vitro fertilization (IVF) in PVA-containing medium had higher penetration rates (P < 0.01) than sperm in BSA-containing medium (54 vs. 11%). The effect of equine preovulatory follicular fluid on bovine oocyte penetration was assessed. Bovine oocytes were matured in tissue culture medium-199 with 0, 20, 50, or 100% equine preovulatory follicular fluid, and 1 IU/mL of equine chorionic gonadotropin. Stallion sperm were treated with 8BrcAMP + ionomycin in PVA- or BSA-containing media. The penetration rates of bovine zona-free oocytes by stallion sperm were again higher with PVA (47%) than BSA (18%; P < 0.01). Penetration rates of oocytes matured in 100% follicular fluid were higher (P < 0.05) than for oocytes matured with 0% follicular fluid. The effects of equine follicular fluid and PVA/BSA during sperm capacitation on standard bovine IVF were examined. Culture of bovine oocytes with equine follicular fluid did not affect oocyte maturation or penetration rates after IVF. Bovine sperm capacitated with heparin in PVA-containing medium yielded lower (P < 0.05) fertilization rates than those capacitated in BSA-containing medium when incubated with both zona-intact and zona-free bovine oocytes. In summary...

Vitrification of immature feline oocytes with a commercial kit for bovine embryo vitrification

Apparicio, M.; Ruggeri, E.; Luvoni, G. C.
Fonte: Universidade Estadual Paulista Publicador: Universidade Estadual Paulista
Tipo: Artigo de Revista Científica Formato: 240-244
ENG
Relevância na Pesquisa
36.99%
The aim of this study was to evaluate the suitability of a commercial kit for bovine embryo vitrification for cryopreserving cat oocytes and to evaluate comparatively the effects of its use with slow freezing procedure on cryotolerance in terms of morphology and oocyte resumption of meiosis. Germinal vesicle stage oocytes isolated from cat ovaries were either vitrified (n=72) using a vitrification kit for bovine embryo or slow frozen (n=69) by exposing oocyte to ethylene glycol solution before being transferred to a programmable embryo freezer. After thawing and warming, oocytes were cultured for 48h and then were examined for meiosis resumption using bisbenzimide fluorescent staining (Hoechst 33342). Fresh immature oocytes (n=92) were used as the control group. The proportion of oocytes recovered in a morphologically normal state after thawing/warming was significantly higher in frozen oocytes (94.5%) than in the vitrified ones (75%, p<0.01). Morphological integrity after culture was similar in vitrified (73.6%) and slow frozen oocytes (76.8%); however, only 37.5% of the morphologically normal oocytes resumed meiosis after vitrification compared to 60.9% of those submitted to slow freezing procedure (p<0.01). Fresh oocytes showed higher morphological integrity (91.3%) and meiosis resumption rates (82.6%...

Maturation of pig oocytes in vitro in a medium with pyruvate

Gonzales-Figueroa,H.; Gonzales-Molfino,H.M.
Fonte: Associação Brasileira de Divulgação Científica Publicador: Associação Brasileira de Divulgação Científica
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/06/2005 EN
Relevância na Pesquisa
36.89%
The aim of in vitro maturation oocyte systems is to produce oocytes of comparable quality to those derived in vivo. The present study was designed to examine the surface morphological changes of the cumulus-oocyte complex (COC) and nuclear maturation in a culture system containing pyruvate. Ovaries were obtained from a slaughterhouseand transported to the laboratory within 2 h at 35-39ºC,and rinsed three times in 0.9% NaCl. The COCs were harvested from the ovaries and in vitro maturation was evaluated in San Marcos (SM) medium, a chemically defined culture system containing 22.3 mM sodium pyruvate. Oocytes were cultured in SM, SM + porcine follicular fluid (pFF) and in SM + pFF + gonadotropins (eCG and hCG) for 20-22 h and then without hormonal supplements for an additional 20-22 h. After culture, the degree of cumulus expansion and frequency of nuclear maturation were determined. Oocytes matured in SM (40.9%) and SM + pFF (42.9%) showed moderate cumulus expansion, whereas oocytes matured in SM + pFF + gonadotropins (54.6%) showed high cumulus expansion. The maturation rate of cultured oocytes, measured in function of the presence of the polar corpuscle, did not differ significantly between SM (40.9 ± 3.6%) and SM + pFF (42.9 ± 3.7%). These results indicate that pig oocytes can be successfully matured in a chemically definedmedium and suggest a possible bifunctional role of pyruvate as an energy substrate and as an antioxidant protecting oocytes against the stress of the in vitro environment.

Vacuum-cooled liquid nitrogen increases the developmental ability of vitrified-warmed bovine oocytes

Santos,Rodrigo Marques dos; Barreta,Marcos Henrique; Frajblat,Marcel; Cucco,Diego Córdova; Mezzalira,Joana Claudia; Bunn,Silvério; Cruz,Fabiano Buss; Vieira,Arnaldo Diniz; Mezzalira,Alceu
Fonte: Universidade Federal de Santa Maria Publicador: Universidade Federal de Santa Maria
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/10/2006 EN
Relevância na Pesquisa
36.95%
The objective of this study was to determine the effects of vacuum-cooled liquid nitrogen on the development of vitrified immature (germinal vesicle stage; GV) and mature (metaphase II; MII) bovine oocytes after re-warming. Liquid nitrogen was exposed to either atmospheric pressure or to a vacuum (300mm Hg for 45sec); the latter decreased the temperature of the liquid nitrogen to -200°C. Partially denuded oocytes were vitrified either just after selection (GV) or after 22 hours of in vitro maturation (MII) in TCM 199 medium + 10% of estrous mare serum. For vitrification, oocytes were firstly exposed to an intermediate solution (10% EG + 10% DMSO) for 30sec, followed by the vitrification solution (20% EG + 20% DMSO + 0.5M sucrose) for 20sec. Groups of three or four oocytes were loaded into an open-pulled-straw and directly plunged into liquid nitrogen. Oocytes were subsequently re-warmed by exposure to air (25°C) for 4sec, followed by 5 min exposure to decreasing concentrations (0.3 and 0.15M) of sucrose. Fertilization (Day 0) was done with 2 x 106 spermatozoa mL-1 (selected by a swim-up procedure) and incubated for 18 to 22 hours. Presumptive zygotes were cultured at 39°C in four-well dishes with SOFaaci medium, under 5% CO2 and saturated humidity. Cleavage (Day 2) and blastocyst rates (Day 8) were 33.9 and 4.2%...

Annexin II mRNA expression in bovine oocytes during follicular development

Costa,Luis Fabiano Santos da; Machado,Márcia Silveira Netto; Oliveira,João Francisco Coelho de; Zamberlan,Gustavo; Gonçalves,Paulo Bayard Dias
Fonte: Sociedade Brasileira de Genética Publicador: Sociedade Brasileira de Genética
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/01/2006 EN
Relevância na Pesquisa
36.95%
We investigated the expression of calcium-dependent phospholipid binding protein annexin-II (Ann-II) messenger RNA (mRNA) during preantral follicle development and in oocytes from antral follicles of different diameters (< 3 mm, 5 to 8 mm and > 8 mm). The action of retinol on Ann-II mRNA expression in mature oocytes was also examined. Only oocytes from secondary preantral follicles expressed Ann-II mRNA and at the germinal vesicle stage expression by oocytes from follicles larger than 8 mm was significantly higher (p < 0.05) compared with oocytes from follicles smaller than 3 mm or between 5 and 8 mm. Ann-II mRNA expression by metaphase II oocytes from follicles larger than 8 mm was significantly higher (p < 0.05) than that from oocytes from follicles smaller than 3 mm, with oocytes from both these size-classes showing similar levels of Ann-II mRNA expression as oocytes recovered from 5-8 mm follicles. In the presence of retinol, Ann-II mRNA expression was higher than when retinol was absent (p < 0.05). Our data indicate that Ann-II mRNA expression is highest in competent oocytes and that retinol increases Ann-II mRNA and may be involved in the regulation of oocyte competence by decreasing the translation and/or degradation of Ann-II mRNA.

Padrão de metilação dos genes IGF2 e XIST em ovócitos oriundos de folículos pré-antrais de vacas Nelore (Bos taurus indicus); Methylation pattern of the IGF2 and XIST genes in oocytes obtained of the preantral follicles from Nellore cows (Bos taurus indicus)

Gomes, Luís Fernando Soares
Fonte: Universidade Federal de Uberlândia Publicador: Universidade Federal de Uberlândia
Tipo: Dissertação
POR
Relevância na Pesquisa
36.91%
Com o intuito de aumentar a produtividade, visando o máximo de aproveitamento do material genético disponível, várias biotecnologias reprodutivas vêm sendo desenvolvidas e utilizadas. Apesar de ovócitos obtidos de folículos pré-antrais não terem ainda competência para produzir embrião, são uma fonte muito importante de gametas a ser utilizada na produção in vitro de embriões. Entender a reprogramação epigenética que acontece nestes ovócitos é necessário quando se vislumbra a possibilidade futura de uso de ovócitos obtidos de folículos pré-antrais para a produção de embriões. Neste trabalho objetivou-se avaliar o padrão de metilação numa DMR do último éxon do gene IGF2 em ovócitos de 65-90 μm e do éxon 1 do gene XIST em ovócitos ≤ 20 μm e 65-90 μm obtidos de folículos pré-antrais. Para o IGF2, os ovócitos de folículos secundários finais de 65-90 μm apresentaram 31,09 ± 31,01% de metilação, e para o XIST, os ovócitos de folículos primordiais ≤ 20 μm e os ovócitos de folículos secundários finais de 65-90 μm apresentaram 15,17 ± 32,82% e 4,6 ± 3,46%, respectivamente. O IGF2 apresentou-se hipometilado e analisando com os dados na literatura, observa-se que esta região do gene sofre um processo de reprogramação epigenética durante a ovogênese. O XIST apresentou um processo de desmetilação no decorrer do desenvolvimento do ovócito e com uma tendência de permanecer até a fecundação. Conclui-se que estas duas regiões genômicas estudadas sofrem um processo de reprogramação epigenética durante a gametogênese. __________________________________________________________________________________________ ABSTRACT; In order to increase productivity looking for maximum use of genetic material available...

Vitrifica????o de o??citos imaturos de eq??inos : caracter??sticas morfol??gicas ultra-estruturais e matura????o nuclear in vitro; Vitrification of immature equine oocytes: ultra structural morphologic characteristics and nuclear in vitro maturation

CURCIO, Bruna da Rosa
Fonte: Universidade Federal de Pelotas; Biotecnologia; Programa de P??s-Gradua????o em Biotecnologia; UFPel; BR Publicador: Universidade Federal de Pelotas; Biotecnologia; Programa de P??s-Gradua????o em Biotecnologia; UFPel; BR
Tipo: Tese de Doutorado Formato: application/pdf
POR
Relevância na Pesquisa
37.03%
The aim of the study was to investigate: 1) the ultrastructural morphologic characteristics in equine oocytes subjected to different times of exposure to cryoprotectant solutions and 2) the effect of initial cumulus morphology and cryoprotectants in the nuclear in vitro maturation (IVM) of the vitrified immature equine oocytes. The oocytes were obtained from ovaries from a slaughterhouse. In the first study 30 oocytes where divided in three groups: Control group (G1, n=10); Group 2 (G2, n=10), the oocytes were vitrified for exposure to VS-1 for 3min and VS-2 for 1min; Group 3 (G3, n=10), exposure to VS1 for 1.5min and VS-2 for 30sec. The oocytes were vitrified in open-pulledstraws (OPS). The ultrastructural characteristics where observed using a transmission electron microscope. The oocytes were classified as: I) oocytes morphologically normal; II) oocytes which presented intermediate damage but had completed organelles, and III) oocytes with severe morphological abnormalities. In the second study, compact (Ccp; n=248) and expanded (Cex; n=264) cumulus oocyte complexes were divided in three groups: Control, Treatment-1 (T1 - Ethylene glycol-EG + Dimetyl sulfoxide-DMSO + SIB) and Treatment-2 (T2 - Formamide + EG + DMSO + Polyvinylpyrrolidone + SIB). The control group was immediately matured in vitro...

Efeito dos m??todos de vitrifica????o, OPS e SSV, com adi????o de bloqueador sint??tico de gelo, sobre a viabilidade de o??citos de camundongos e bovinos; Effect of the vitrification methods OPS and SSV, with inclusion of a synthetic ice blocker, on the viability of mice and bovine oocytes

SANTOS, Elisa Caroline da Silva
Fonte: Universidade Federal de Pelotas; Biotecnologia; Programa de P??s-Gradua????o em Biotecnologia; UFPel; BR Publicador: Universidade Federal de Pelotas; Biotecnologia; Programa de P??s-Gradua????o em Biotecnologia; UFPel; BR
Tipo: Dissertação Formato: application/pdf
POR
Relevância na Pesquisa
36.89%
Oocyte vitrification is a valuable tool for preservation of genetic material. This study compared the effects of vitrification in OPS and SSV, with addition of SupercoolTM X- 1000 (copolymer), on the viability of mature murine oocytes and immature bovine oocytes. Oocytes were vitrified in OPS and SSV, with addition of 0.1%, 1.0% copolymer and without copolymer, besides a control group with no vitrification. Murine oocytes were evaluated for membrane viability, in the first experiment, and for cleavage rate, in the second experiment. Results were superior with the concentration of 0.1% copolymer, for both methods, in the first experiment. In the second experiment, the SSV method without copolymer presented lower cleavage rate (9.2%) than the control group (26.6%). In the third experiment, bovine oocytes were vitrified and evaluated for maturation and membrane viability, but the results were numerically inferior than those for the control group, for both methods. Those results indicate that both vitrification methods can be used with inclusion of 0.1% of copolymer, for mature murine oocytes, considering membrane viability, but the SSV method without copolymer should not be used due to its low cleavage rate. However, the procedures tested in this study are not recommended for cryopreservation of bovine oocytes.; A vitrifica????o de o??citos ?? uma metodologia valiosa para a conserva????o de material gen??tico. Este trabalho comparou o efeito dos m??todos de vitrifica????o OPS e SSV...

Developmental competence of in vivo and in vitro matured porcine oocytes after subzonal sperm injection

Nagashima, H.; Grupen, C.; Ashman, R.; Nottle, M.
Fonte: WILEY-LISS Publicador: WILEY-LISS
Tipo: Artigo de Revista Científica
Publicado em //1996 EN
Relevância na Pesquisa
36.98%
In vivo and in vitro matured porcine oocytes were fertilized by subzonal sperm injection (SUZI), and their subsequent development in vitro was examined to determine whether ooplasmic incompetence is the major cause of limited developmental ability of in vitro matured/ fertilized porcine oocytes (Experiment 1). There was no significant difference in rates of fertilization (61% vs. 70%), monospermy (37% vs. 45%), and male pronuclear formation (77% vs. 61%) between in vivo and in vitro matured oocytes. Blastocyst formation rate was significantly lower for in vitro matured oocytes (11% vs. 42%; P < 0.001). Forty-six percent of in vivo matured oocytes cleaved to the 2-4 cell stage by 24 hr in culture after SUZI, compared with 3% of in vitro matured oocytes (P < 0.01). In experiment 2, in vitro development of in vitro matured oocytes with evenly and unevenly granulated cytoplasm were compared after SUZI to examine whether developmentally competent in vitro matured oocytes can be identified on the basis of morphological appearance. Most of the blastocysts obtained developed from oocytes with unevenly granulated cytoplasm (7/56 vs. 1/45; P > 0.05). Experiment 3 revealed that the proportion of oocytes with evenly granulated cytoplasm was originally low (11%) in the population of oocytes used for in vitro maturation...

DNA double strand breaks but not interstrand crosslinks prevent progress through meiosis in fully grown mouse oocytes

Yuen, W.S.; Merriman, J.A.; O'Bryan, M.K.; Jones, K.T.
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
Publicado em //2012 EN
Relevância na Pesquisa
36.95%
There is some interest in how mammalian oocytes respond to different types of DNA damage because of the increasing expectation of fertility preservation in women undergoing chemotherapy. Double strand breaks (DSBs) induced by ionizing radiation and agents such as neocarzinostatin (NCS), and interstrand crosslinks (ICLs) induced by alkylating agents such as mitomycin C (MMC), are toxic DNA lesions that need to be repaired for cell survival. Here we examined the effects of NCS and MMC treatment on oocytes collected from antral follicles in mice, because potentially such oocytes are readily collected from ovaries and do not need to be in vitro grown to achieve meiotic competency. We found that oocytes were sensitive to NCS, such that this ionizing radiation mimetic blocked meiosis I and caused fragmented DNA. In contrast, MMC had no impact on the completion of either meiosis I or II, even at extremely high doses. However, oocytes treated with MMC did show γ-H2AX foci and following their in vitro maturation and parthenogenetic activation the development of the subsequent embryos was severely compromised. Addition of MMC to 1-cell embryos caused a similarly poor level of development, demonstrating oocytes have eventual sensitivity to this ICL-inducing agent but this does not occur during their meiotic division. In oocytes...

Effect of Hormonal Supplementation Periods and In Vitro Maturation Media on Developmental Competence of Pig Oocytes

LIMA, Alecssandra Sobreira de; MARQUES, Mariana Groke; NASCIMENTO, Anibal Ballarotti; ASSUMPCAO, Mayra Elena Ortiz D'Avila; VISITIN, Jose Antonio
Fonte: UNIV FED RIO GRANDE DO SUL Publicador: UNIV FED RIO GRANDE DO SUL
Tipo: Artigo de Revista Científica
ENG
Relevância na Pesquisa
37%
Background: The oocyte ability to undergo successful fertilization, cleavage and embryonic development depends on meiotic maturation and developmental competence acquisition. In vitro maturation (IVM) protocols currently use eCG, hCG or a combination of both, the effect of these gonadotrophins during IVM and subsequent embryonic development is still controversial. Several media have been used for IVM of porcine oocytes: TCM199, Whitten's and NCSU23 have also been shown to support pig oocyte IVM. This study was designed to determine the effect of hormonal supplementation period and maturation media during in vitro maturation of pig oocytes (1) and subsequent embryonic development (2). Materials, Methods & Results: Oocytes with intact cumulus oophurus layers and homogeneous cytoplasm were collected from prebubertal gilts. IVM was subjected in NCSU23, TCM199 or Whitten's media supplemented with 10 IU/mL eCG and 10 IU/mL hCG for the first 24 or 48 h of IVM. In each replicate the oocytes were fixed every 4 h from 32 to 48 h IVM or the past 48 h after IVM, oocytes were fertilized in vitro in mTBM medium for six hours and cultured in NCSU23 medium for nine days. Cleavage, blastocyst and hatching rates were evaluated at 48 h (day 2), 168 h (day 7) and 216 h (day 9)...

Anomalias meióticas de oócitos de pacientes com endometriose submetidas à estimulação ovariana; Meiotic abnormalities of oocytes from patients with endometriosis submitted to ovarian stimulation

BARCELOS, Ionara Diniz Evangelista Santos; VIEIRA, Rodolpho Cruz; FERREIRA, Elisa Melo; ARAÚJO, Maria Cristina Picinato Medeiros de; MARTINS, Wellington de Paula; FERRIANI, Rui Alberto; NAVARRO, Paula Andrea de Albuquerque Salles
Fonte: Federação Brasileira das Sociedades de Ginecologia e Obstetrícia Publicador: Federação Brasileira das Sociedades de Ginecologia e Obstetrícia
Tipo: Artigo de Revista Científica
POR
Relevância na Pesquisa
36.95%
OBJETIVO: avaliar o fuso meiótico e a distribuição cromossômica de oócitos maturados in vitro, obtidos de ciclos estimulados de mulheres inférteis com endometriose e fatores masculino e/ou tubário de infertilidade (Grupo Controle), comparando as taxas de maturação in vitro (MIV) entre os dois grupos avaliados. MÉTODOS: quatorze pacientes com endometriose e oito com fator tubário ou masculino, submetidas à estimulação ovariana para injeção intracitoplasmática de espermatozóide, foram selecionadas, prospectiva e consecutivamente, e constituíram os Grupos de Estudo e Controle, respectivamente. Oócitos imaturos (46 e 22, respectivamente, dos Grupos Endometriose e Controle) foram submetidos à MIV. Oócitos que apresentaram a extrusão do primeiro corpúsculo polar foram fixados e corados para avaliação dos microtúbulos e cromatina por técnica de imunofluorescência. A análise estatística foi realizada utilizando o teste exato de Fisher, com significância estatística quando p<0,05. RESULTADOS: não se observou diferença significativa nas taxas de MIV entre os dois grupos avaliados (45,6 e 54,5%, respectivamente, nos Grupos Endometriose e Controle). A organização cromossômica e do fuso meiótico foi observada em 18 e 11 oócitos dos Grupos Endometriose e Controle...

Análise citogenética de oócitos de jaguatirica (Leopardus pardalis) e gato-do-mato-pequeno (Leopardus tigrinus) coletados após estimulação ovariana; Cytogenetic analyses of ocelot (Leopardus pardalis) and tigrinus (Leopardus tigrinus) oocytes collected after ovarian stimulation

PAZ, Regina Celia Rodrigues da; ADANIA, Cristina Harumi; BARNABÉ, Valquíria Hyppólito; BARNABE, Renato Campanarut
Fonte: São Paulo Publicador: São Paulo
Tipo: Artigo de Revista Científica
POR
Relevância na Pesquisa
36.89%
Este estudo representa a primeira avaliação da maturação nuclear de oócitos por análise citogenética realizada em duas espécies de felídeos brasileiros ameaçados de extinção: L. pardalis (n=5) e L. tigrinus (n=4). Os animais foram submetidos à estimulação ovariana alternada com eCG-hCG e pFSH-pLH a cada quatro meses pelo período de dois anos, perfazendo um total de 6 intervenções. Os oócitos foram recuperados por vídeo-laparoscopia, caracterizados quanto à morfologia e utilizados para determinação dos estágios do ciclo meiótico por análise citogenética e maturação pela caracterização de metáfase II. Dos 33 oócitos de jaguatirica avaliados 12% (n=4) apresentaram cromossomos condensados em seu interior e dos 11 oócitos de gato-do-mato-pequeno avaliados 36% (n=4) apresentaram cromossomos condensados em seu interior, no entanto, nenhum oócito encontrava-se em metáfase II. Avaliação morfológica dos oócitos mostrou as mesmas características encontradas em outros mamíferos. Comparando os tratamentos, não houve diferença significativa (p>0,05) no número total de estruturas ovarianas (folículos e corpos lúteos recentes) observadas em estimulações alternadas sucessivas, nas duas espécies estudadas. Também não houve diferença significativa em relação ao total de estruturas ovarianas encontradas em cada tratamento (5...

Effect of Roscovitine on nuclear maturation, MPF and MAP kinase activity and embryo development of prepubertal goat oocytes

Jiménez y de Macedo, Ana Raquel; Izquierdo Tugas, Maria Dolors; Urdaneta Vargas, Aixa Efrailda; Anguita Bustamante, Begoña; Paramio Nieto, María Teresa
Fonte: Universidade Autônoma de Barcelona Publicador: Universidade Autônoma de Barcelona
Tipo: info:eu-repo/semantics/article; info:eu-repo/semantics/publishedVersion; info:eu-repo/semantics/acceptedVersion Formato: application/pdf
Publicado em //2006 ENG
Relevância na Pesquisa
37%
The low number of embryos obtained from IVM–IVF–IVC of prepubertal goat oocytes could be due to an incomplete cytoplasmic maturation. Roscovitine (ROS) inhibits MPF and MAP kinase activity and maintains the oocyte at Germinal Vesicle (GV) stage. The aim of this study was to determine if meiotic activity is arrested in prepubertal goat oocytes cultured with 0, 12.5, 25, 50 and 100 m M of ROS for 24 h. A group of oocytes from adult goats was cultured with 25 m M of ROS to compare the effect of ROS on prepubertal and adult goat oocytes. A sample of oocytes was stained to evaluate the nuclear stage at oocyte collection time and after ROS incubation. IVM-oocytes not exposed to ROS formed the control group. Prepubertal goat IVM-oocytes were inseminated and cultured for 8 days. The percentage of oocytes at GV stage, after exposition to ROS was significantly higher in adult goat oocytes (64.5%) than in prepubertal goat oocytes. No differences were found among 25, 50 and 100 m M ROS concentrations (29, 23 and 26%, oocytes at GV stage, respectively). After 8 days of culture, no differences in total embryos were observed between control oocytes and oocytes treated with 12.5 and 25 m M (45.2, 36.1 and 39.4%, respectively), however the percentage of lastocysts was higher in the control group. Western blot for the MAPK and p34 cdc2 showed that both enzymes were active in prepubertal goat oocytes after 24 h of ROS exposition. In conclusion...