title: In vivo oocyte IGF-1 priming increases inner cell mass proliferation of in vitro-formed bovine blastocysts.
abstract: Studies addressing the effects of supraphysiological levels of IGF-1 on oocyte developmental competence are relevant for unravelling conditions resulting in high bioavailability of IGF-1, such as the polycystic ovary syndrome (PCOS). This study investigated the effects of supraphysiological levels of IGF-1 during in vivo folliculogenesis on the morula-blastocyst transition in bovine embryos. Compacted morulae were non-surgically collected and frozen for subsequent mRNA expression analysis (IGF1R, IGBP3, TP53, AKT1, SLC2A1, SLC2A3, and SLC2A8), or underwent confocal microscopy analysis for protein localization (IGF1R and TP53), or were cultured in vitro for 24 h. In vitro-formed blastocysts were subjected to differential cell staining. The mRNA expression of SLC2A8 was higher in morulae collected from cows treated with IGF-1. Both IGF1R and TP53 protein were present in the plasma membrane and cytoplasm. IGF-1 treatment did not affect protein localization of both IGF1R and TP53. In vitro-formed blastocysts derived from morulae recovered from IGF-1-treated cows displayed a higher number of cells in the inner cell mass (ICM). Total cell number (TCN) of in vitro-formed blastocysts was not affected. A higher mean ICM/TCN proportion was observed in in vitro-formed blastocysts derived from morulae collected from cows treated with IGF-1. The percentage of in vitro-formed blastocysts displaying a low ICM/TCN proportion was decreased by IGF-1 treatment. In vitro-formed blastocysts with a high ICM/TCN proportion were only detected in IGF-1 treated cows. Results show that even a short in vivo exposure of oocytes to a supraphysiological IGF-1 microenvironment can increase ICM cell proliferation in vitro during the morula to blastocyst transition.
authors.affiliation: Institute of Farm Animal Genetics, Biotechnology, Friedrich-Loeffler-Institut, Höltystraße 10, Mariensee, 31535 Neustadt, Germany.
authors.forename: M A
authors.initials: MA
authors.lastname: Velazquez
authors.forename: K-G
authors.initials: KG
authors.lastname: Hadeler
authors.forename: D
authors.initials: D
authors.lastname: Herrmann
authors.forename: W A
authors.initials: WA
authors.lastname: Kues
authors.forename: B
authors.initials: B
authors.lastname: Rémy
authors.forename: J-F
authors.initials: JF
authors.lastname: Beckers
authors.forename: H
authors.initials: H
authors.lastname: Niemann
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000737:chemistry / Q000166:cytology*; D002417:Cattle / Q000196:embryology*; D049109:Cell Proliferation / Q000187:drug effects; D046149:Embryo Culture Techniques / Q000662:veterinary; D047108:Embryonic Development / Q000235:genetics; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D015870:Gene Expression / Q000187:drug effects; D051246:Glucose Transport Proteins, Facilitative / Q000235:genetics; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D009028:Morula / Q000737:chemistry / Q000166:cytology; D009865:Oocytes / Q000187:drug effects*; D010053:Ovary / Q000187:drug effects; D012333:RNA, Messenger / Q000032:analysis; D017526:Receptor, IGF Type 1 / Q000032:analysis / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000032:analysis / Q000235:genetics
chemical_list: D051246:Glucose Transport Proteins, Facilitative; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53; D007334:Insulin-Like Growth Factor I; D017526:Receptor, IGF Type 1