Journal
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY
Volume 166, Issue 1, Pages 81-90Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.cbpb.2013.07.005
Keywords
Ovarian development; Salmonid; Vitellogenin; Vitellogenin receptor; Yolk formation
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Funding
- JSPS KAKENHI [24-1961, 23580243]
- MEXT KAKENHI [22380103]
- Japan Science and Technology Agency [AS231Z00518E]
- Grants-in-Aid for Scientific Research [22380103, 23580242, 23580243] Funding Source: KAKEN
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A cDNA encoding a vitellogenin receptor with 8 ligand binding repeats (vtgr) was cloned from ovaries of the cutthroat trout, Oncorhynchus clarki. In situ hybridization and quantitative PCR analyses revealed that the main site of vtgr mRNA expression was the oocytes. Expression was strongly detected in perinucleous stage oocytes, gradually decreased as oocytes grew, and became hardly detectable in vitellogenic oocytes. A rabbit antibody (a-Vtgr) was raised against a recombinant Vtgr protein in order to immunologically detect and localize Vtgr within the ovarian follicles. Western blotting using a-Vtgr detected a bold band with an apparent mass of similar to 95-105 kDa in an ovarian preparation that also bound Sakhalin taimen, Hucho perryi, vitellogenin in ligand blots. Immunohistochemistry using a-Vtgr revealed that the Vtgr was uniformly distributed throughout the ooplasm of perinucleolus stage oocytes, subsequently translocated to the periphery of lipid droplet stage oocytes, and became localized to the oolemma during vitellogenesis. We provide the first characterization of Vtgr at both the transcriptional and the translational levels in the cutthroat trout, and our results suggest that this receptor is involved in uptake of Vtg by oocytes of this species. (C) 2013 Elsevier Inc. All rights reserved.
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