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1.
Theriogenology ; 71(6): 939-46, 2009 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-19150733

RESUMO

In vitro maturation (IVM) of mammalian oocytes does not support the same rates of embryo development or pregnancy when compared to oocytes that have matured in vivo. Therefore, environment has a significant influence on the oocyte's ability to complete maturation and acquire the mRNA and proteins required for successful fertilization and normal embryonic development. The aim of this study was to analyze the MII oocyte transcriptome between in vivo and in vitro conditions. Total RNA was extracted, processed and hybridized to the Affymetrix GeneChip Bovine Genome Array. Following normalization of the microarray data, analysis revealed 10 differentially expressed genes after IVM compared to in vivo matured controls, including Aqp3, Sept7, Abhd4 and Siah2 (P<0.05). K-means cluster analysis coupled with associated gene ontology, identified several biological processes affected by IVM, including metabolism, energy pathways, cell organization and biogenesis, and cell growth and maintenance. Quantitative real-time PCR validated the microarray data and also revealed altered expression levels after IVM of specific putatively imprinted genes, Igf2r, Peg3 and Snrpn (P<0.05). Distinct IVM transcription patterns reflected the oocyte's response to its surrounding environment. Monitoring transcription levels of key oocyte maturation genes may subsequently assist in improving IVM success.


Assuntos
Bovinos , Perfilação da Expressão Gênica/veterinária , Oócitos/química , Oócitos/crescimento & desenvolvimento , Animais , Metabolismo dos Carboidratos , Metabolismo Energético , Feminino , Técnicas In Vitro , Análise de Sequência com Séries de Oligonucleotídeos/veterinária , Oócitos/metabolismo , Reação em Cadeia da Polimerase/veterinária
2.
Theriogenology ; 61(7-8): 1215-23, 2004 May.
Artigo em Inglês | MEDLINE | ID: mdl-15036956

RESUMO

Transportation of equine ovaries would allow shipment of oocytes for research purposes or transfer after the death of a valuable mare. The objective of this study was to compare two temperatures for maintaining ovaries during a transport interval of 18-24 h. The goal was to obtain pregnancies after transport of ovaries, maturation of oocytes in vitro, and transfer of oocytes. Each shipment was composed of ovaries four to seven mares collected from an abattoir. From each mare, one ovary was packaged at approximately 12 degrees C, and the other was packaged at approximately 22 degrees C. Upon arrival at our laboratory, oocytes were collected and cultured for 24 h. For each transfer, between 9 and 15 oocytes from each group were placed into the oviducts of estrous mares through standing flank laparotomies. Recipients received human chorionic gonadotropin (hCG; 2000 IU, i.v.) 30-36 h before transfer (to synchronize ovulation). Recipients were inseminated 18-20 h before transfers with 2 x 10(9) progressively motile sperm. Uteri of recipients were examined with ultrasound to determine the number of developing embryos. On Day 16 ( ovulation = day 0), developing embryos were recovered by uterine lavage. Parentage verification was performed on recovered vesicles. Pregnancy rates were analyzed by Chi-square. The percentage of oocytes that developed into embryonic vesicles on Day 16 was not different between transport temperatures (22 degrees C, 13/73, 18% versus 12 degrees C, 11/73, 15%). In conclusion, pregnancies were obtained from in vitro matured oocytes that were recovered from ovaries transported for 18-24h at 12 or 22 degrees C.


Assuntos
Cavalos , Oócitos/fisiologia , Ovário/fisiologia , Manejo de Espécimes/veterinária , Temperatura , Animais , Feminino , Doação de Oócitos/veterinária , Preservação de Órgãos/veterinária , Ovário/citologia , Gravidez , Manejo de Espécimes/métodos , Fatores de Tempo
3.
Theriogenology ; 61(4): 705-13, 2004 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-14698059

RESUMO

The objectives were to compare embryo development rates after oocyte transfer with: (1) intrauterine or intraoviductal inseminations of fresh semen versus intraoviductal insemination of frozen semen; (2) intraoviductal versus intrauterine inseminations of cooled semen. In Experiment I, oocytes were transferred into the oviduct, and recipients were inseminated into the uterus with 1 x 10(9) fresh spermatozoa, or into the oviduct with 2 x 10(5) fresh or frozen-thawed spermatozoa. In Experiment II, semen was cooled to 5 degrees C before intrauterine insemination with 2 x 10(9) spermatozoa or intraoviductal inseminations of 2 x 10(5) spermatozoa (deposited with the oocytes). In Experiment I, embryo development rates were similar (P>0.05) for intrauterine versus intraoviductal inseminations when fresh semen was used (8/14, 57% and 9/11, 82%, respectively). However, embryo development rates were lower (P<0.05) when frozen spermatozoa were placed within the oviduct (1/12, 8%). In Experiment II, embryo development rates were higher (P<0.05) when cooled semen was used for intrauterine (19/23, 83%) versus intraoviductal (4/16, 25%) inseminations. We concluded that intraoviductal insemination can be successfully performed using fresh spermatozoa. However, the use of cooled and frozen spermatozoa for intraoviductal inseminations was less successful, and needs further investigation.


Assuntos
Cavalos , Inseminação Artificial/veterinária , Doação de Oócitos/veterinária , Preservação do Sêmen/veterinária , Animais , Temperatura Baixa , Criopreservação , Desenvolvimento Embrionário e Fetal , Tubas Uterinas , Feminino , Inseminação Artificial/métodos , Masculino , Doação de Oócitos/métodos , Gravidez , Preservação do Sêmen/métodos , Útero
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