title: Identification of stably expressed reference genes for RT-qPCR data normalization in defined localizations of cyclic bovine ovaries.
abstract: Ovaries are highly complex organs displaying morphological, molecular and functional differences between their cortical zona parenchymatosa and medullary zona vasculosa, and also between the different cyclic luteal stages. Objective of the present study was to validate expression stability of twelve putative reference genes (RGs) in bovine ovaries, considering the intrinsic heterogeneity of bovine ovarian tissue with regard to different luteal stages and intra-ovarian localizations. The focus was on identifying RGs, which are suitable to normalize RT-qPCR results of ovaries collected from clinical healthy cattle, irrespective of localization and the hormonal stage. Expression profiles of twelve potential reference genes (GAPDH, ACTB, YWHAZ, HPRT1, SDHA, UBA52, POLR2C, RPS9, ACTG2, H3F3B, RPS18 and RPL19) were analysed. Evaluation of gene expression differences was performed using genorm, normfinder, and bestkeeper software. The most stably expressed genes according to genorm, normfinder and bestkeeper approaches contained the candidates H3F3B, RPS9, YWHAZ, RPS18, POLR2C and UBA52. Of this group, the genes YWHAZ, H3F3B and RPS9 could be recommended as best-suited RGs for normalization purposes on healthy bovine ovaries irrespective of the luteal stage or intra-ovarian localization.
authors.affiliation: Department of Veterinary Medicine, Institute of Veterinary Anatomy, Freie Universität Berlin, Koserstraße 20, 14195, Berlin, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Schoen
authors.affiliation: Department of Veterinary Medicine, Institute of Veterinary Anatomy, Freie Universität Berlin, Koserstraße 20, 14195, Berlin, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Plendl
authors.affiliation: Department of Veterinary Medicine, Institute of Veterinary Biochemistry, Freie Universität Berlin, Oertzenweg 19b, 14163, Berlin, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Gabler
authors.affiliation: Department of Veterinary Medicine, Institute of Veterinary Anatomy, Freie Universität Berlin, Koserstraße 20, 14195, Berlin, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Kaessmeyer
mesh_terms: D000003:Abattoirs; D000465:Algorithms; D000818:Animals; D002417:Cattle / Q000033:anatomy & histology / Q000235:genetics*; D003338:Corpus Luteum / Q000502:physiology; D018076:DNA, Complementary / Q000378:metabolism; D030762:Estrous Cycle / Q000235:genetics; D005260:Female; D005786:Gene Expression Regulation / Q000235:genetics*; D010053:Ovary / Q000033:anatomy & histology / Q000502:physiology*; D011247:Pregnancy; D012313:RNA / Q000302:isolation & purification; D012015:Reference Standards; D012269:Ribosomal Proteins / Q000235:genetics*; D012984:Software
chemical_list: D018076:DNA, Complementary; D012269:Ribosomal Proteins; D012313:RNA