PUBLICATION

Molecular cloning, regulation, and functional analysis of two GHS-R genes in zebrafish

Authors
Eom, J., Hong, A., Kang, Y.H., Yoo, H.J., Chang, E.J., Kang, S.W., Yoon, S.Y., Kim, S.Y., Song, Y.
ID
ZDB-PUB-140615-7
Date
2014
Source
Experimental cell research   326(1): 10-21 (Journal)
Registered Authors
Keywords
Ca(2+) oscillation, GHS-R, Ghrelin, Zebrafish
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Blotting, Western
  • Calcium/metabolism*
  • Cloning, Molecular
  • Fluorescent Antibody Technique
  • Gene Expression Regulation/drug effects*
  • Ghrelin/pharmacology*
  • HEK293 Cells
  • Humans
  • Molecular Sequence Data
  • Protein Isoforms
  • RNA, Messenger/genetics
  • Rats
  • Real-Time Polymerase Chain Reaction
  • Receptors, Ghrelin/genetics
  • Receptors, Ghrelin/metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Homology, Amino Acid
  • Signal Transduction
  • Zebrafish
PubMed
24928276 Full text @ Exp. Cell Res.
Abstract
Mammalian ghrelin is derived from stomach and regulates growth hormone release and appetite by modulating GHS-R (Growth Hormone Secretagogue Receptor) activity. Zebrafish has been developed as a forward genetic screening model system and previous screening identified a number of genes involved multiple signaling pathways. In this system, ghrelin has been identified and its function and regulation has been shown to be highly conserved to that of mammals. Here, we identified three isoforms of zGHS-R1 and one zGHS-R2 (zGHS-R2a), and characterized their expression, regulation and function.. Three isoforms of zGHS-R1, which we named zGHS-R1a, zGHS-R1b, and zGHS-R1c, are generated by alternative splicing. The expression of zGHS-R1 is highly enriched in brain, intestine tissue, and skin tissues. Compared to zGHS-R1, the expression pattern of zGHS-R2a is rather evenly distributed. A 15 day fasting elevated expression of zGHS-R1 and zGHS-R2 transcripts in anterior intestine tissues, but not in brain. Whereas zGHS-R1a, zGHS-R1c, and zGHS-R2a appear to be presented on the plasma membrane, the localization of zGHS-R1b seems to be restricted in the intracellular region. Treatment of ghrelin agonist, L692,585 or goldfish ghrelin peptides but not rat ghrelin, elevated intracellular Ca(2+) level and phosphorylation of ERK in HEK-293 cells expressing zGHS-R1a., but not zGHS-R1b, zGHS-R1c, or zGHS-R2a. It appears that besides core ghrelin peptide sequence of GS/TSF additional amino acids are required for the activation of zGHS-R1a, as rat ghrelin induces neither intracellular Ca(2+) mobilization nor ERK phosphrylation. These results suggest that ghrelin system in zebrafish is highly conserved to that of mammals, and thus is an ideal model in vivo model for dissecting ghrelin system.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping