title: Early cleavages influence the molecular and the metabolic pattern of individually cultured bovine blastocysts.
abstract: Embryo morphokinetics suggests that the timing of the first embryonic cell divisions may predict the developmental potential of an embryo; however, correlations between embryonic morphokinetics and physiology are not clear. Here, we used RNA sequencing to determine the gene expression profile of in vitro-produced early- and late-dividing bovine embryos and their respective blastocysts, and compared these profiles to in vivo-produced blastocysts to identify differentially expressed genes (DEGs). Principal component analysis revealed that fast- and slow-dividing embryos possess similar transcript abundance over the first cleavages. By the blastocyst stage, however, more DEGs were observed between the fast- and slow-dividing embryo groups, whereas blastocysts from the slow-dividing group were more similar to in vivo-produced blastocysts. Gene ontology enrichment analysis showed that the slow-dividing and in vivo-produced blastocysts shared biological processes related to groups of up- or down-regulated genes when compared to the fast-dividing blastocysts. Based on these DEG results, we characterized the relationship between developmental kinetics and energy metabolism of in vitro-produced bovine embryos. Embryos from fast- and slow-dividing groups exhibited different pyruvate and lactate metabolism at 22 hr post-in vitro culture (hpc), glucose consumption at 96 hpc, and glutamate metabolism at 168 hpc. Glycogen storage was similar between cleavage-stage and morulae groups, but was higher in the blastocysts of the slow-dividing group. On the other hand, blastocysts of the fast-dividing group had a higher concentration of lipids. Taken together, these data identify transcriptomic and metabolic differences between embryos with different morphokinetics, suggesting that sorting embryos based on cleavage speed may select for different metabolic patterns. Mol. Reprod. Dev. 83: 324-336, 2016. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Center of Natural and Human Sciences, Universidade Federal do ABC, Santo Andre, Sao Paulo, Brazil.
authors.forename: Marcella Pecora
authors.initials: MP
authors.lastname: Milazzotto
authors.affiliation: Department of Animal Reproduction, University of São Paulo, Sao Paulo, Brazil.
authors.forename: Marcelo Demarchi
authors.initials: MD
authors.lastname: Goissis
authors.affiliation: Department of Animal Science, University of California, Davis, California.
authors.forename: James Lee
authors.initials: JL
authors.lastname: Chitwood
authors.affiliation: Center of Natural and Human Sciences, Universidade Federal do ABC, Santo Andre, Sao Paulo, Brazil.
authors.forename: Kelly
authors.initials: K
authors.lastname: Annes
authors.affiliation: Center of Natural and Human Sciences, Universidade Federal do ABC, Santo Andre, Sao Paulo, Brazil.
authors.forename: Carlos Alexandre
authors.initials: CA
authors.lastname: Soares
authors.affiliation: Center of Natural and Human Sciences, Universidade Federal do ABC, Santo Andre, Sao Paulo, Brazil.
authors.forename: Jéssica
authors.initials: J
authors.lastname: Ispada
authors.affiliation: Department of Animal Reproduction, University of São Paulo, Sao Paulo, Brazil.
authors.forename: Mayra Elena Ortiz Ávila
authors.initials: ME
authors.lastname: Assumpção
authors.affiliation: Department of Animal Science, University of California, Davis, California.
authors.forename: Pablo Juan
authors.initials: PJ
authors.lastname: Ross
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism*; D002417:Cattle; D002455:Cell Division; D002970:Cleavage Stage, Ovum; D003470:Culture Media / Q000378:metabolism; D048749:Cytokinesis; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D015870:Gene Expression; D011247:Pregnancy; D025341:Principal Component Analysis; D012333:RNA, Messenger; D013997:Time Factors; D046509:Tissue Culture Techniques; D059467:Transcriptome
chemical_list: D003470:Culture Media; D012333:RNA, Messenger