Molecular cloning and functional analysis of the zebrafish follicle-stimulating hormone (FSH)beta promoter

Comp Biochem Physiol B Biochem Mol Biol. 2010 Feb;155(2):155-63. doi: 10.1016/j.cbpb.2009.10.017.

Abstract

In the present study, we cloned and characterized a zebrafish follicle-stimulating hormone (zfFSH)beta promoter with deletion fragments transfected into a tilapia ovary (TO2) cell line, and demonstrated that the zfFSHbeta promoter responded to 6h of gonadotropin-releasing hormone (GnRH) treatment by activating calcium influx and protein kinase C (PKC), but after 24h, GnRH induction was generated by activation of extracellular-regulated kinase (ERK)1/2 and repression by PKC. Furthermore, to study the promoter-specific expression, we constructed a series of FSHbeta (4.0-, 3.0-, 2.0-, and 1.0-kb) promoter-driven green fluorescent protein (GFP) fragments encoding the GFP complementary DNA transgene which was microinjected into zebrafish embryos. Morphological studies of transgenic zebrafish indicated that the FSHbeta promoter-driven GFP transcripts appeared in the heart, skin, and vertebrae.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Bioreactors
  • Calcium / pharmacology
  • Cell Line
  • Cloning, Molecular
  • Embryo, Nonmammalian / metabolism
  • Enzyme Activation
  • Female
  • Fertilization
  • Follicle Stimulating Hormone, beta Subunit / genetics*
  • Follicle Stimulating Hormone, beta Subunit / metabolism
  • Gene Expression Regulation / drug effects
  • Gonadotropin-Releasing Hormone / metabolism
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Mitogen-Activated Protein Kinase 3 / metabolism
  • Ovary / cytology
  • Ovary / drug effects
  • Ovary / metabolism
  • Ovum / physiology
  • Pituitary Gland / metabolism
  • Promoter Regions, Genetic / genetics*
  • Protein Kinase C / metabolism
  • Signal Transduction
  • Time Factors
  • Zebrafish / embryology
  • Zebrafish / genetics*
  • Zebrafish / physiology

Substances

  • Follicle Stimulating Hormone, beta Subunit
  • Gonadotropin-Releasing Hormone
  • Protein Kinase C
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • Calcium