PUBLICATION

Molecular Cloning and Functional Characterization of a Zebrafish Nuclear Progesterone Receptor

Authors
Chen, S.X., Bogerd, J., García-López, A., de Jonge, H., de Waal, P.P., Hong, W.S., and Schulz, R.W.
ID
ZDB-PUB-090914-45
Date
2010
Source
Biology of reproduction   82(1): 171-181 (Journal)
Registered Authors
Bogerd, Jan
Keywords
nuclear progesterone receptor, steroid release, gonad development, testis
MeSH Terms
  • Animals
  • Cloning, Molecular
  • Hormone Antagonists
  • Hydroxyprogesterones/metabolism*
  • Male
  • Mifepristone
  • Open Reading Frames
  • RNA, Messenger/metabolism
  • Receptors, Progesterone/metabolism*
  • Sequence Analysis, DNA
  • Spermatogenesis*
  • Testis/metabolism*
  • Testosterone/analogs & derivatives
  • Transcriptional Activation
  • Zebrafish/embryology
  • Zebrafish/metabolism*
(all 16)
PubMed
19741208 Full text @ Biol. Reprod.
Abstract
Progestagenic sex steroid hormones play critical roles in reproduction across vertebrates, including teleost fish. To further our understanding of how progesterone modulates testis functions in fish, we set out to clone a progesterone receptor (pgr) cDNA exhibiting nuclear hormone receptor features from zebrafish testis. The open-reading frame of pgr consists of 1854 bp, coding for a 617 amino acids long protein showing highest similarity with other piscine Pgr proteins. Functional characterization of the receptor expressed in mammalian cells revealed that zebrafish Pgr exhibited progesterone-specific, dose-dependent induction of reporter gene expression, with 17alpha,20beta-dihydroxy- 4-pregnen-3-one (DHP), a typical piscine progesterone, showing the highest potency. Expression of pgr mRNA: (i) appeared in embryos at 8 h post fertilization; (ii) was significantly higher in developing ovary than in early transforming testis at 4 wk of age but vice versa in young adults at 12 wk of age and thus resembling the expression pattern of the germ cell marker piwil1; and, (iii) was restricted to Leydig and Sertoli cells in adult testis. Zebrafish testicular explants released DHP concentration-dependently in response to high concentrations of recombinant zebrafish gonadotropins. In addition, DHP stimulated 11-ketotestosterone release from zebrafish testicular explants, but only in the presence of its immediate precursor 11beta-hydroxytestosterone. This stimulatory activity was blocked by a Pgr antagonist (RU486), suggesting that 11beta-hydroxysteroid dehydrogenase activity was stimulated by DHP via Pgr. Our data suggest that DHP contributes to the regulation of Leydig cell steroidogenesis and potentially - via Sertoli cells - also of germ cell differentiation in zebrafish testis.
Genes / Markers
Marker Marker Type Name
ddx4GENEDEAD (Asp-Glu-Ala-Asp) box polypeptide 4
pgrGENEprogesterone receptor
piwil1GENEpiwi-like RNA-mediated gene silencing 1
sycp3GENEsynaptonemal complex protein 3
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Expression
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
hu2479
    Point Mutation
    zf45TgTransgenic Insertion
      1 - 2 of 2
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      Human Disease / Model
      No data available
      Sequence Targeting Reagents
      No data available
      Fish
      Antibodies
      Orthology
      Engineered Foreign Genes
      Marker Marker Type Name
      EGFPEFGEGFP
      1 - 1 of 1
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      Mapping