Embryonic Development and Gene Expression in Oocytes Cultured In Vitro in Supplemented Pre-Maturation and Maturation Media

This study aimed at assessing the effect of the addition of brain-derived neurotrophic factor (10 ng/ml BDNF) and/or cysteamine (100 μm CYS) during pre-maturation and BDNF, CYS or leptin (10 ng/ml LEP) during maturation culture in vitro on embryo development and oocyte gene expression in cattle. Ooc...

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Bibliographic Details
Published in:	Reproduction in domestic animals Vol. 46; no. 1; pp. e31 - e38
Main Authors:	Adona, PR, de Bem, THC, Mesquita, LG, Rochetti, RC, Leal, CLV
Format:	Journal Article
Language:	English
Published:	Oxford, UK Blackwell Publishing Ltd 01-02-2011
Subjects:	Animal reproduction Animals Brain-Derived Neurotrophic Factor - administration & dosage Cattle Cattle - embryology Cell culture Cells, Cultured Culture Media Cysteamine - administration & dosage Embryo Culture Techniques - veterinary Embryology Embryonic Development - drug effects Female Fertilization in Vitro - veterinary Gene expression Gene Expression - drug effects Leptin - administration & dosage Oocytes - drug effects Oocytes - growth & development Oocytes - metabolism Polymerase Chain Reaction Reproductive technologies
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Summary:	This study aimed at assessing the effect of the addition of brain-derived neurotrophic factor (10 ng/ml BDNF) and/or cysteamine (100 μm CYS) during pre-maturation and BDNF, CYS or leptin (10 ng/ml LEP) during maturation culture in vitro on embryo development and oocyte gene expression in cattle. Oocytes were obtained by the aspiration of 2- to 8-mm follicles from slaughtered cows. In Experiment 1, oocytes were pre-matured for 24 h with 10 μm butyrolactone I in the presence or not of BDNF and/or CYS followed by in vitro maturation (IVM), fertilization (IVF) and culture (IVC). In Experiment 2, oocytes were submitted to IVM with BDNF, CYS or LEP or no supplements followed by IVF and IVC. In Experiment 3, oocytes were pre-matured with BDNF and CYS followed by IVM or only in vitro matured with BDNF. Samples for quantitative PCR (qPCR) were collected after pre-maturation (BGV) and after IVM of pre-matured oocytes (BMII) or immediately after follicle aspiration (immature control = GV) and IVM (matured control = MII). Embryo production was not affected by the inclusion of the different factors either during pre-maturation or maturation culture (∼43% blastocysts, p > 0.05). Transcripts analysis showed that most genes (NLRP5, ZAR1, GPX1, KEAP1, SPHK2, HSP70 and PSMP1) were downregulated (p < 0.05) after IVM irrespective of being previously pre-matured. The relative abundance of BAX, BCL2, IGFBP3 and ARFRP1 transcripts was unaffected by pre-maturation or maturation (p > 0.05). In conclusion, supplementation of in vitro pre-maturation (BDNF and/or CYS) or maturation media (BDNF, CYS or LEP) did not improve embryo development. Gene expression was not affected by pre-maturation treatment, but some genes were downregulated after maturation, probably related to selective and differential degradation.
Bibliography:	http://dx.doi.org/10.1111/j.1439-0531.2010.01618.x ark:/67375/WNG-CB47PXS7-B istex:D6BD9FD7B68D280899EF8675654065F234A84C15 ArticleID:RDA1618 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0936-6768 1439-0531
DOI:	10.1111/j.1439-0531.2010.01618.x