Inhibitory guanine-nucleotide-binding-regulatory protein a subunits in medaka (Oryzias latipes) oocytes. cDNA cloning and decreased expression of proteins during oocyte maturation

Yuichi Oba, Michiyasu Yoshikuni, Minoru Tanaka, Masatoshi Mita, Yoshitaka Nagahama

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

We have previously shown that pertussis-toxin-sensitive inhibitory guanine-nucleotide-binding-regulatory proteins (G proteins) are involved in the signal transduction of steroidal maturation-inducing hormone (MIH) of rainbow trout (Oncorhynchus mykiss) oocytes, 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-DP) [Yoshikuni, M. and Nagahama, Y. (1994) Dev. Biol. 166, 615-622]. In this study, we obtained five different cDNA fragments of G protein α subunits from medaka (Oryzias latipes) intact ovarian follicles (three subtypes of G(iα), G(iαa), G(iαb) and G(iαc); two subtypes of G(sα), G(sαd), and G(sαe)). Using a newly developed extraction method for medaka oocyte RNA, we demonstrated that oocytes expressed both G(iαa) and G(iαc), but not G(iαb). Full-length cDNA clones for G(iαa) and G(iαc) were then isolated from a medaka ovarian follicle cDNA library. The predicted amino acid sequences of G(iαa) and G(iαc) exhibited significant similarity with G(iα1) and G(iα2) of other species, respectively. Both G(iαa) and G(iαc) possessed a specific Cys residue in the C-terminal region that was the site for ADP-ribosylation by pertussis toxin. G(oα) another G protein that is ADP-ribosylated by pertussis toxin, was not detected in oocytes, although it was expressed in brain tissue. Western blot analyses using a specific antibody against G(iα1) and G(iα2) subunit proteins revealed that in both medaka and rainbow trout G(iα) subunit protein (40 kDa) contents were abundant in plasma membranes of postvitellogenic immature oocytes, decreased in mature oocytes, and were absent in ovulated eggs. Furthermore, specific 17α,20β-DP binding to plasma membranes was higher in postvitellogenic immature oocytes than in ovulated eggs. Taken together, these results suggest that G(iαa) and/or G(iαc) may be involved in the transduction of the signal from 17α,20β-DP receptors during oocyte maturation of fish oocytes.

Original languageEnglish
Pages (from-to)846-853
Number of pages8
JournalEuropean Journal of Biochemistry
Volume249
Issue number3
DOIs
Publication statusPublished - Jan 1 1997

Fingerprint

Oryzias
Guanine Nucleotides
Cloning
Pertussis Toxin
Protein Subunits
GTP-Binding Proteins
Oocytes
Organism Cloning
Carrier Proteins
Complementary DNA
Cell membranes
Adenosine Diphosphate
Signal transduction
Proteins
Oncorhynchus mykiss
Gene Library
Fish
Brain
Ovarian Follicle
Hormones

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Inhibitory guanine-nucleotide-binding-regulatory protein a subunits in medaka (Oryzias latipes) oocytes. cDNA cloning and decreased expression of proteins during oocyte maturation. / Oba, Yuichi; Yoshikuni, Michiyasu; Tanaka, Minoru; Mita, Masatoshi; Nagahama, Yoshitaka.

In: European Journal of Biochemistry, Vol. 249, No. 3, 01.01.1997, p. 846-853.

Research output: Contribution to journalArticle

@article{2d5576f16fb14a78b14ab36bab345f57,
title = "Inhibitory guanine-nucleotide-binding-regulatory protein a subunits in medaka (Oryzias latipes) oocytes. cDNA cloning and decreased expression of proteins during oocyte maturation",
abstract = "We have previously shown that pertussis-toxin-sensitive inhibitory guanine-nucleotide-binding-regulatory proteins (G proteins) are involved in the signal transduction of steroidal maturation-inducing hormone (MIH) of rainbow trout (Oncorhynchus mykiss) oocytes, 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-DP) [Yoshikuni, M. and Nagahama, Y. (1994) Dev. Biol. 166, 615-622]. In this study, we obtained five different cDNA fragments of G protein α subunits from medaka (Oryzias latipes) intact ovarian follicles (three subtypes of G(iα), G(iαa), G(iαb) and G(iαc); two subtypes of G(sα), G(sαd), and G(sαe)). Using a newly developed extraction method for medaka oocyte RNA, we demonstrated that oocytes expressed both G(iαa) and G(iαc), but not G(iαb). Full-length cDNA clones for G(iαa) and G(iαc) were then isolated from a medaka ovarian follicle cDNA library. The predicted amino acid sequences of G(iαa) and G(iαc) exhibited significant similarity with G(iα1) and G(iα2) of other species, respectively. Both G(iαa) and G(iαc) possessed a specific Cys residue in the C-terminal region that was the site for ADP-ribosylation by pertussis toxin. G(oα) another G protein that is ADP-ribosylated by pertussis toxin, was not detected in oocytes, although it was expressed in brain tissue. Western blot analyses using a specific antibody against G(iα1) and G(iα2) subunit proteins revealed that in both medaka and rainbow trout G(iα) subunit protein (40 kDa) contents were abundant in plasma membranes of postvitellogenic immature oocytes, decreased in mature oocytes, and were absent in ovulated eggs. Furthermore, specific 17α,20β-DP binding to plasma membranes was higher in postvitellogenic immature oocytes than in ovulated eggs. Taken together, these results suggest that G(iαa) and/or G(iαc) may be involved in the transduction of the signal from 17α,20β-DP receptors during oocyte maturation of fish oocytes.",
author = "Yuichi Oba and Michiyasu Yoshikuni and Minoru Tanaka and Masatoshi Mita and Yoshitaka Nagahama",
year = "1997",
month = "1",
day = "1",
doi = "10.1111/j.1432-1033.1997.00846.x",
language = "English",
volume = "249",
pages = "846--853",
journal = "FEBS Journal",
issn = "1742-464X",
publisher = "Wiley-Blackwell",
number = "3",

}

TY - JOUR

T1 - Inhibitory guanine-nucleotide-binding-regulatory protein a subunits in medaka (Oryzias latipes) oocytes. cDNA cloning and decreased expression of proteins during oocyte maturation

AU - Oba, Yuichi

AU - Yoshikuni, Michiyasu

AU - Tanaka, Minoru

AU - Mita, Masatoshi

AU - Nagahama, Yoshitaka

PY - 1997/1/1

Y1 - 1997/1/1

N2 - We have previously shown that pertussis-toxin-sensitive inhibitory guanine-nucleotide-binding-regulatory proteins (G proteins) are involved in the signal transduction of steroidal maturation-inducing hormone (MIH) of rainbow trout (Oncorhynchus mykiss) oocytes, 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-DP) [Yoshikuni, M. and Nagahama, Y. (1994) Dev. Biol. 166, 615-622]. In this study, we obtained five different cDNA fragments of G protein α subunits from medaka (Oryzias latipes) intact ovarian follicles (three subtypes of G(iα), G(iαa), G(iαb) and G(iαc); two subtypes of G(sα), G(sαd), and G(sαe)). Using a newly developed extraction method for medaka oocyte RNA, we demonstrated that oocytes expressed both G(iαa) and G(iαc), but not G(iαb). Full-length cDNA clones for G(iαa) and G(iαc) were then isolated from a medaka ovarian follicle cDNA library. The predicted amino acid sequences of G(iαa) and G(iαc) exhibited significant similarity with G(iα1) and G(iα2) of other species, respectively. Both G(iαa) and G(iαc) possessed a specific Cys residue in the C-terminal region that was the site for ADP-ribosylation by pertussis toxin. G(oα) another G protein that is ADP-ribosylated by pertussis toxin, was not detected in oocytes, although it was expressed in brain tissue. Western blot analyses using a specific antibody against G(iα1) and G(iα2) subunit proteins revealed that in both medaka and rainbow trout G(iα) subunit protein (40 kDa) contents were abundant in plasma membranes of postvitellogenic immature oocytes, decreased in mature oocytes, and were absent in ovulated eggs. Furthermore, specific 17α,20β-DP binding to plasma membranes was higher in postvitellogenic immature oocytes than in ovulated eggs. Taken together, these results suggest that G(iαa) and/or G(iαc) may be involved in the transduction of the signal from 17α,20β-DP receptors during oocyte maturation of fish oocytes.

AB - We have previously shown that pertussis-toxin-sensitive inhibitory guanine-nucleotide-binding-regulatory proteins (G proteins) are involved in the signal transduction of steroidal maturation-inducing hormone (MIH) of rainbow trout (Oncorhynchus mykiss) oocytes, 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-DP) [Yoshikuni, M. and Nagahama, Y. (1994) Dev. Biol. 166, 615-622]. In this study, we obtained five different cDNA fragments of G protein α subunits from medaka (Oryzias latipes) intact ovarian follicles (three subtypes of G(iα), G(iαa), G(iαb) and G(iαc); two subtypes of G(sα), G(sαd), and G(sαe)). Using a newly developed extraction method for medaka oocyte RNA, we demonstrated that oocytes expressed both G(iαa) and G(iαc), but not G(iαb). Full-length cDNA clones for G(iαa) and G(iαc) were then isolated from a medaka ovarian follicle cDNA library. The predicted amino acid sequences of G(iαa) and G(iαc) exhibited significant similarity with G(iα1) and G(iα2) of other species, respectively. Both G(iαa) and G(iαc) possessed a specific Cys residue in the C-terminal region that was the site for ADP-ribosylation by pertussis toxin. G(oα) another G protein that is ADP-ribosylated by pertussis toxin, was not detected in oocytes, although it was expressed in brain tissue. Western blot analyses using a specific antibody against G(iα1) and G(iα2) subunit proteins revealed that in both medaka and rainbow trout G(iα) subunit protein (40 kDa) contents were abundant in plasma membranes of postvitellogenic immature oocytes, decreased in mature oocytes, and were absent in ovulated eggs. Furthermore, specific 17α,20β-DP binding to plasma membranes was higher in postvitellogenic immature oocytes than in ovulated eggs. Taken together, these results suggest that G(iαa) and/or G(iαc) may be involved in the transduction of the signal from 17α,20β-DP receptors during oocyte maturation of fish oocytes.

UR - http://www.scopus.com/inward/record.url?scp=0030664897&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030664897&partnerID=8YFLogxK

U2 - 10.1111/j.1432-1033.1997.00846.x

DO - 10.1111/j.1432-1033.1997.00846.x

M3 - Article

VL - 249

SP - 846

EP - 853

JO - FEBS Journal

JF - FEBS Journal

SN - 1742-464X

IS - 3

ER -