title: Validation of reference genes for ovarian tissue from capuchin monkeys (Cebus apella).
abstract: There is no tradition in studies reporting the effect of exposure to cryoprotectants or simply hypoxia and hypothermia on gene expression in the ovarian tissue and there has been only one study on reference or target genes quantification, and comparisons of normoxic with hypoxic, hypothermic and toxic conditions. Our aim in the present study was to investigate the stability of three reference genes in the ovarian tissue of capuchin monkeys (Cebus apella). To this end, fresh and cryoprotectant-exposed ovarian biopsies were used. Both fresh and exposed ovarian tissues were subjected to total RNA extraction and synthesis of cDNA. cDNA was amplified by real-time polymerase chain reaction (PCR), and GeNorm, BestKeeper and NormFinder software were used to evaluate the stability of glyceraldehyde-2-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyltransferase 1 (HPRT1) and TATA-binding protein (TBP). Results demonstrated that, in the ovarian tissue from capuchin monkeys, HPRT1 and TBP were the most suitable reference genes and thus could be used as parameters to normalize data in future studies. In contrast, GAPDH appeared as the least stable gene among the tested reference genes. In conclusion, HPRT1 and TBP were the most stable reference genes in fresh and cryoprotectant-exposed ovarian tissue from capuchin monkeys.
authors.affiliation: Laboratory of Wild Animal Biology and Medicine, Universidade Federal do Pará, Brazil.
authors.forename: A B
authors.initials: AB
authors.lastname: Brito
authors.forename: J S
authors.initials: JS
authors.lastname: Lima
authors.forename: D C
authors.initials: DC
authors.lastname: Brito
authors.forename: L N
authors.initials: LN
authors.lastname: Santana
authors.forename: N N
authors.initials: NN
authors.lastname: Costa
authors.forename: M S
authors.initials: MS
authors.lastname: Miranda
authors.forename: O M
authors.initials: OM
authors.lastname: Ohashi
authors.forename: R R
authors.initials: RR
authors.lastname: Santos
authors.forename: S F S
authors.initials: SF
authors.lastname: Domingues
mesh_terms: D000818:Animals; D002428:Cebus; D003451:Cryoprotective Agents / Q000494:pharmacology; D005260:Female; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D007035:Hypothermia; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D010053:Ovary / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010100:Oxygen / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction; D035181:TATA-Box Binding Protein / Q000235:genetics*
chemical_list: D003451:Cryoprotective Agents; D012333:RNA, Messenger; D035181:TATA-Box Binding Protein; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D007041:Hypoxanthine Phosphoribosyltransferase; D010100:Oxygen