Expression and function of cGMP-dependent protein kinase type I during medaka fish embryogenesis

Takehiro Yamamoto, Norio Suzuki

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

We isolated and characterized cDNA clones (PKG Iα and PKG Iβ) for medaka fish cGMP-dependent protein kinase (PKG) Iα and Iβ, and demonstrated that both are expressed in the embryos after late gastrula stage. Whole-mount in situ hybridization using each isoform-specific probe revealed that the transcripts of the PKG Ia gene were present in the spinal cord and gill arch, whereas those of the PXG Iβ gene were only weakly expressed in these organs, but highly expressed in the otic vesicles. Injection of PKG Iα-specific morpholino antisense oligonucleotides (Iα-MO) into two-cell stage medaka fish embryos caused severe abnormalities in the developing embryos, such as the development of a hammer-like head, fusion of the developing eyes, and degeneration of cells around the eyes, whereas injection of PKG Iβ-specific morpholino antisense oligonucleotides (Iβ-MO) caused fewer abnormalities in the embryos, even when injected at higher concentrations than Iα-MO. The PKG I-overexpressing embryos exhibited smaller eyes and enlargement of the forebrain, a pheno-type similar to that observed in the cAMP-dependent protein kinase (PKA)-depressed embryos. In the PKG-deficient embryos, a sonic hedgehog (shh)-target gene, HNF-3β, was expressed weakly, and this phenotype was similar to that observed in the PKA-overexpressing embryos suggesting that the cGMP/PKG signaling pathway is involved in some steps of shh signaling. We also demonstrated that Gli proteins, shh-downstream molecules, are phosphorylated by the NO/cGMP signaling pathway, probably by PKG in NG108-15 neuroblastoma cells. These results imply that PKG and PKA share common substrates and work in an opposite manner during the early embryogenesis of medaka fish.

Original languageEnglish
Pages (from-to)16979-16986
Number of pages8
JournalJournal of Biological Chemistry
Volume280
Issue number17
DOIs
Publication statusPublished - 2005 Apr 29

Fingerprint

Cyclic GMP-Dependent Protein Kinase Type I
Oryzias
Morpholinos
Antisense Oligonucleotides
Fish
Embryonic Development
Fishes
Embryonic Structures
Genes
Protein Kinases
Hedgehog Proteins
Hedgehogs
Hammers
Arches
Cyclic AMP-Dependent Protein Kinases
Protein Isoforms
Fusion reactions
Complementary DNA
Molecules
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Expression and function of cGMP-dependent protein kinase type I during medaka fish embryogenesis. / Yamamoto, Takehiro; Suzuki, Norio.

In: Journal of Biological Chemistry, Vol. 280, No. 17, 29.04.2005, p. 16979-16986.

Research output: Contribution to journalArticle

@article{2bfdfbda464a48b09913c68bd909ac58,
title = "Expression and function of cGMP-dependent protein kinase type I during medaka fish embryogenesis",
abstract = "We isolated and characterized cDNA clones (PKG Iα and PKG Iβ) for medaka fish cGMP-dependent protein kinase (PKG) Iα and Iβ, and demonstrated that both are expressed in the embryos after late gastrula stage. Whole-mount in situ hybridization using each isoform-specific probe revealed that the transcripts of the PKG Ia gene were present in the spinal cord and gill arch, whereas those of the PXG Iβ gene were only weakly expressed in these organs, but highly expressed in the otic vesicles. Injection of PKG Iα-specific morpholino antisense oligonucleotides (Iα-MO) into two-cell stage medaka fish embryos caused severe abnormalities in the developing embryos, such as the development of a hammer-like head, fusion of the developing eyes, and degeneration of cells around the eyes, whereas injection of PKG Iβ-specific morpholino antisense oligonucleotides (Iβ-MO) caused fewer abnormalities in the embryos, even when injected at higher concentrations than Iα-MO. The PKG I-overexpressing embryos exhibited smaller eyes and enlargement of the forebrain, a pheno-type similar to that observed in the cAMP-dependent protein kinase (PKA)-depressed embryos. In the PKG-deficient embryos, a sonic hedgehog (shh)-target gene, HNF-3β, was expressed weakly, and this phenotype was similar to that observed in the PKA-overexpressing embryos suggesting that the cGMP/PKG signaling pathway is involved in some steps of shh signaling. We also demonstrated that Gli proteins, shh-downstream molecules, are phosphorylated by the NO/cGMP signaling pathway, probably by PKG in NG108-15 neuroblastoma cells. These results imply that PKG and PKA share common substrates and work in an opposite manner during the early embryogenesis of medaka fish.",
author = "Takehiro Yamamoto and Norio Suzuki",
year = "2005",
month = "4",
day = "29",
doi = "10.1074/jbc.M412433200",
language = "English",
volume = "280",
pages = "16979--16986",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "17",

}

TY - JOUR

T1 - Expression and function of cGMP-dependent protein kinase type I during medaka fish embryogenesis

AU - Yamamoto, Takehiro

AU - Suzuki, Norio

PY - 2005/4/29

Y1 - 2005/4/29

N2 - We isolated and characterized cDNA clones (PKG Iα and PKG Iβ) for medaka fish cGMP-dependent protein kinase (PKG) Iα and Iβ, and demonstrated that both are expressed in the embryos after late gastrula stage. Whole-mount in situ hybridization using each isoform-specific probe revealed that the transcripts of the PKG Ia gene were present in the spinal cord and gill arch, whereas those of the PXG Iβ gene were only weakly expressed in these organs, but highly expressed in the otic vesicles. Injection of PKG Iα-specific morpholino antisense oligonucleotides (Iα-MO) into two-cell stage medaka fish embryos caused severe abnormalities in the developing embryos, such as the development of a hammer-like head, fusion of the developing eyes, and degeneration of cells around the eyes, whereas injection of PKG Iβ-specific morpholino antisense oligonucleotides (Iβ-MO) caused fewer abnormalities in the embryos, even when injected at higher concentrations than Iα-MO. The PKG I-overexpressing embryos exhibited smaller eyes and enlargement of the forebrain, a pheno-type similar to that observed in the cAMP-dependent protein kinase (PKA)-depressed embryos. In the PKG-deficient embryos, a sonic hedgehog (shh)-target gene, HNF-3β, was expressed weakly, and this phenotype was similar to that observed in the PKA-overexpressing embryos suggesting that the cGMP/PKG signaling pathway is involved in some steps of shh signaling. We also demonstrated that Gli proteins, shh-downstream molecules, are phosphorylated by the NO/cGMP signaling pathway, probably by PKG in NG108-15 neuroblastoma cells. These results imply that PKG and PKA share common substrates and work in an opposite manner during the early embryogenesis of medaka fish.

AB - We isolated and characterized cDNA clones (PKG Iα and PKG Iβ) for medaka fish cGMP-dependent protein kinase (PKG) Iα and Iβ, and demonstrated that both are expressed in the embryos after late gastrula stage. Whole-mount in situ hybridization using each isoform-specific probe revealed that the transcripts of the PKG Ia gene were present in the spinal cord and gill arch, whereas those of the PXG Iβ gene were only weakly expressed in these organs, but highly expressed in the otic vesicles. Injection of PKG Iα-specific morpholino antisense oligonucleotides (Iα-MO) into two-cell stage medaka fish embryos caused severe abnormalities in the developing embryos, such as the development of a hammer-like head, fusion of the developing eyes, and degeneration of cells around the eyes, whereas injection of PKG Iβ-specific morpholino antisense oligonucleotides (Iβ-MO) caused fewer abnormalities in the embryos, even when injected at higher concentrations than Iα-MO. The PKG I-overexpressing embryos exhibited smaller eyes and enlargement of the forebrain, a pheno-type similar to that observed in the cAMP-dependent protein kinase (PKA)-depressed embryos. In the PKG-deficient embryos, a sonic hedgehog (shh)-target gene, HNF-3β, was expressed weakly, and this phenotype was similar to that observed in the PKA-overexpressing embryos suggesting that the cGMP/PKG signaling pathway is involved in some steps of shh signaling. We also demonstrated that Gli proteins, shh-downstream molecules, are phosphorylated by the NO/cGMP signaling pathway, probably by PKG in NG108-15 neuroblastoma cells. These results imply that PKG and PKA share common substrates and work in an opposite manner during the early embryogenesis of medaka fish.

UR - http://www.scopus.com/inward/record.url?scp=20444439930&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=20444439930&partnerID=8YFLogxK

U2 - 10.1074/jbc.M412433200

DO - 10.1074/jbc.M412433200

M3 - Article

C2 - 15710621

AN - SCOPUS:20444439930

VL - 280

SP - 16979

EP - 16986

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 17

ER -