| Title: | Identification and characterization of lipocalin-type prostaglandin D(2) synthase homologs in the urine of male rockfish |
| Author(s): | Yamaguchi Y; Namgung J; Nagata J; Kawasaki T; Hara A; Todo T; Hiramatsu N; |
| Address: | "Graduate School of Fisheries Sciences, Hokkaido University, 3-1-1 Minato, Hakodate, Hokkaido 041-8611, Japan. Education & Research Group for Future Strategy of Aquatic Life Industry, Jeju National University, Jeju 63243, Republic of Korea. Division of Marine Life Science, Faculty of Fisheries Sciences, Hokkaido University, 3-1-1 Minato, Hakodate, Hokkaido 041-8611, Japan. Mariculture Fisheries Research Institute, Fisheries Research Department, Hokkaido Research Organization, 1-156-3 Funami, Muroran, Hokkaido 051-0013, Japan. Division of Marine Life Science, Faculty of Fisheries Sciences, Hokkaido University, 3-1-1 Minato, Hakodate, Hokkaido 041-8611, Japan. Electronic address: naoshi@fish.hokudai.ac.jp" |
| DOI: | 10.1016/j.gene.2022.147093 |
| ISSN/ISBN: | 1879-0038 (Electronic) 0378-1119 (Linking) |
| Abstract: | "Black rockfish (Sebastes schlegelii) and its relatives are viviparous marine fish. Males produce urinary proteins during the copulation season; however, the identity of these proteins was unknown. In this study, we focused on high-molecular-weight urinary proteins (HMWups) in male black rockfish. The HMWups were identified by liquid chromatography and tandem mass spectrometry (LC-MS/MS) of urine. In silico analyses of RNA-seq data predicted the tissue distribution of candidate HMWup transcripts and their gene structures. Candidate cDNAs were cloned and a recombinant protein of a major candidate was prepared. Western blotting of urine using an antiserum against the recombinant protein was performed to reconfirm the LC-MS/MS results. Quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry were employed to validate the prediction by RNA-seq and identify the cells producing HMWups, respectively. LC-MS/MS, in conjunction with Western blotting and cDNA cloning, identified the HMWups as lipocalin-type prostaglandin D(2) synthase (l-PGDS) homologs. RNA-seq analyses and qRT-PCR revealed that the l-PGDS homolog transcripts were dominantly expressed in the testis and male kidney; Sertoli cells and epithelial cells in the renal tubules were immunoreactive. These results indicated that major protein components in the urine of male black rockfish are l-PGDS homologs, potentially produced by the renal tubules in the kidney. Male rockfish (genus Sebastes) are thought to release unknown pheromone substances during mating behavior. The knowledge and tools obtained in this study empower research into the role(s) of HMWups in pheromone systems underlying rockfish reproduction. No protein-type teleost pheromone has heretofore been discovered" |
| Keywords: | "Animals Male Chromatography, Liquid Tandem Mass Spectrometry *Perciformes/genetics *Bass Recombinant Proteins Lipocalins/genetics Prostaglandins Gene expression Immuno-biochemistry Lipocalin-type prostaglandin D(2) synthase Recombinant protein Rockfish Ur;" |
| Notes: | "MedlineYamaguchi, Yo Namgung, Jin Nagata, Jun Kawasaki, Takuma Hara, Akihiko Todo, Takashi Hiramatsu, Naoshi eng Netherlands 2022/12/09 Gene. 2023 Feb 20; 854:147093. doi: 10.1016/j.gene.2022.147093. Epub 2022 Dec 5" |
