title: Fine selection of up-regulated genes during ovulation byin vivoinduction of oocyte maturation and ovulation in zebrafish.
abstract: BACKGROUND
  Two essential processes, oocyte maturation and ovulation, are independently induced, but proceed cooperatively as the final step in oogenesis before oocytes become fertilizable. Although these two processes are induced by the same maturation-inducing steroid, 17α, 20β-dihydroxy-4-pregnen-3-one (17, 20β-DHP), in zebrafish, it has been suggested that the receptor, and thus the signal transduction pathway is different for each process. Although much progress has been made in understanding the molecular mechanisms underlying the induction of oocyte maturation, the mechanisms for inducing ovulation remain under investigation. In the present study,in vivoinduction techniques that permit the induction of oocyte maturation and ovulation in living zebrafish (in vivoassays) were used to select highly up-regulated genes (genes associated with ovulation). Using anin vivoassay, ovarian tissues that induced only oocyte maturation could be obtained. This made it possible for the first time to distinguish maturation-inducing genes from ovulation-inducing genes. Using a genome-wide microarray of zebrafish sequences, the gene expression levels were compared among an ethanol (EtOH)-treated group (non-activated group), a diethylstilbestrol (DES)- or testosterone (Tes)-treated group (maturation-induced group), and a 17, 20β-DHP-treated group (maturation- and ovulation-induced group). Ovulation-specific up-regulated genes were selected. The mRNA expression levels of the selected genes were measured by quantitative polymerase chain reaction (qPCR).
  
  
  RESULTS
  Among 34 genes identified, three that showed ovulation-specific increases were selected as candidates potentially associated with ovulation. The ovulation-specific up-regulation of three candidates, slc37a4a, zgc:65811 and zgc:92184 was confirmed by qPCR.
  
  
  CONCLUSIONS
  Ourin vivoassay provides a new approach to precisely select genes associated with ovulation.
authors.affiliation: Integrated Bioscience Section, Graduate School of Science and Technology, National University Corporation Shizuoka University, Ohya 836, Suruga-ku, Shizuoka 422-8529 Japan.
authors.forename: Wanlada
authors.initials: W
authors.lastname: Klangnurak
authors.affiliation: Integrated Bioscience Section, Graduate School of Science and Technology, National University Corporation Shizuoka University, Ohya 836, Suruga-ku, Shizuoka 422-8529 Japan.|Department of Biology, Faculty of Science, National University Corporation Shizuoka University, Shizuoka, 422-8529 Japan.
authors.forename: Toshinobu
authors.initials: T
authors.lastname: Tokumoto
keywords: Oocyte maturation; Ovulation; Steroids; Zebrafish