title: Cyclic AMP Affects Oocyte Maturation and Embryo Development in Prepubertal and Adult Cattle.
abstract: High cAMP levels during in vitro maturation (IVM) have been related to improved blastocyst yields. Here, we employed the cAMP/cGMP modulators, forskolin, IBMX, and cilostamide, during IVM to unravel the role of high cAMP in early embryonic development produced from prepubertal and adult bovine oocytes. Oocytes were collected via transvaginal aspiration and randomly assigned to three experimental groups: TCM24 (24 h IVM/control), cAMP30 (2 h pre-IVM (forskolin-IBMX), 30 h IVM-cilostamide), and DMSO30 (Dimethyl Sulfoxide/vehicle control). After IVM, oocytes were fertilized in vitro and zygotes were cultured in vitro to blastocysts. Meiotic progression, cAMP levels, mRNA abundance of selected genes and DNA methylation were evaluated in oocytes. Blastocysts were used for gene expression or DNA methylation analyses. Blastocysts from the cAMP30 groups were transferred to recipients. The cAMP elevation delayed meiotic progression, but developmental rates were not increased. In immature oocytes, mRNA abundance of PRKACA was higher for cAMP30 protocol and no differences were found for PDE3A, SMAD2, ZAR1, PRDX1 and SLC2A8. EGR1 gene was up-regulated in prepubertal cAMP30 immature oocytes and down-regulated in blastocysts from all in vitro treatments. A similar gene expression profile was observed for DNMT3b, BCL2L1, PRDX1 and SLC2A8 in blastocysts. Satellite DNA methylation profiles were different between prepubertal and adult oocytes and blastocysts derived from the TCM24 and DMSO30 groups. Blastocysts obtained from prepubertal and adult oocytes in the cAMP30 treatment displayed normal methylation profiles and produced offspring. These data indicate that cAMP regulates IVM in prepubertal and adult oocytes in a similar manner, with impact on the establishment of epigenetic marks and acquisition of full developmental competency.
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institut, Mariensee, Germany.|Facultad de Ciencias Agropecuarias, Universidad de Ciencias Aplicadas y Ambientales, Bogotá, Colombia.
authors.forename: Sandra Milena
authors.initials: SM
authors.lastname: Bernal-Ulloa
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institut, Mariensee, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Heinzmann
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institut, Mariensee, Germany.
authors.forename: Doris
authors.initials: D
authors.lastname: Herrmann
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institut, Mariensee, Germany.
authors.forename: Klaus-Gerd
authors.initials: KG
authors.lastname: Hadeler
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institut, Mariensee, Germany.
authors.forename: Patrick
authors.initials: P
authors.lastname: Aldag
authors.affiliation: DNA Sequencing Facility, Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.
authors.forename: Sylke
authors.initials: S
authors.lastname: Winkler
authors.affiliation: DNA Sequencing Facility, Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.
authors.forename: Dorit
authors.initials: D
authors.lastname: Pache
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institut, Mariensee, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Baulain
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institut, Mariensee, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Lucas-Hahn
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institut, Mariensee, Germany.
authors.forename: Heiner
authors.initials: H
authors.lastname: Niemann
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D002417:Cattle; D002452:Cell Count; D018899:CpG Islands / Q000235:genetics; D000242:Cyclic AMP / Q000378:metabolism*; D019175:DNA Methylation; D047108:Embryonic Development; D005260:Female; D018507:Gene Expression Regulation, Developmental; D008540:Meiosis; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism; D011627:Puberty; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D000242:Cyclic AMP