Modification of secondary head-forming activity of microinjected Δβ-catenin mRNA by co-injected spermine and spermidine in Xenopus early embryos

Takamichi Mishina, Kota Fuchimukai, Kazuei Igarashi, Kosuke Tashiro, Koichiro Shiokawa

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Polyamines are essential for cell growth and differentiation. In Xenopus early embryos, per embryo level of spermine is extremely low as compared with that of spermidine. To disclose the possible function of polyamines in Xenopus early embryos, we tested the effect of co-injection of spermine and spermidine on the functioning of exogenously microinjected in vitro-synthesized, Δβ-catenin mRNA which is known to induce a secondary head after being microinjected into a ventral vegetal blastomere in 8-celled Xenopus embryos. Microinjection of Δβ-catenin mRNA in fact induced a secondary axis with a secondary head, and co-injection of spermine or spermidine suppresses induction of the secondary head and secondary axis without drastic effects like induction of immediate cell death or execution of apoptosis at blastula stage. The inhibitory effects were dosage dependent, and at lower doses the inhibition was mainly on secondary head formation rather than on secondary axis formation. We performed similar experiments using GFP mRNA and confirmed that expression of GFP mRNA was also suppressed by co-injection of spermine. We mixed Δβ-catenin mRNA with different amounts of spermine and performed electrophoresis on agarose gels, with a finding that the prior mixing greatly suppressed mRNA migration. These results suggest that an excess amount of spermine as well as spermidine exerts inhibitory effects on mRNA translation, and that the inhibition may be due to direct binding of polyamines to mRNA and a reduction of negative charges on mRNA molecules that might also induce the formation of cross link-like networks among mRNAs.

Original languageEnglish
Pages (from-to)791-801
Number of pages11
JournalAmino Acids
Volume42
Issue number2-3
DOIs
Publication statusPublished - Feb 1 2012

Fingerprint

Catenins
Spermidine
Spermine
Xenopus
Embryonic Structures
Head
Messenger RNA
Polyamines
Injections
Blastula
Blastomeres
Agar Gel Electrophoresis
Microinjections
Protein Biosynthesis
Cell growth
Cell death
Electrophoresis
Cell Differentiation
Sepharose
Cell Death

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Clinical Biochemistry
  • Organic Chemistry

Cite this

Modification of secondary head-forming activity of microinjected Δβ-catenin mRNA by co-injected spermine and spermidine in Xenopus early embryos. / Mishina, Takamichi; Fuchimukai, Kota; Igarashi, Kazuei; Tashiro, Kosuke; Shiokawa, Koichiro.

In: Amino Acids, Vol. 42, No. 2-3, 01.02.2012, p. 791-801.

Research output: Contribution to journalArticle

Mishina, Takamichi ; Fuchimukai, Kota ; Igarashi, Kazuei ; Tashiro, Kosuke ; Shiokawa, Koichiro. / Modification of secondary head-forming activity of microinjected Δβ-catenin mRNA by co-injected spermine and spermidine in Xenopus early embryos. In: Amino Acids. 2012 ; Vol. 42, No. 2-3. pp. 791-801.
@article{62b875ee30ac417fb9e821b3daa80579,
title = "Modification of secondary head-forming activity of microinjected Δβ-catenin mRNA by co-injected spermine and spermidine in Xenopus early embryos",
abstract = "Polyamines are essential for cell growth and differentiation. In Xenopus early embryos, per embryo level of spermine is extremely low as compared with that of spermidine. To disclose the possible function of polyamines in Xenopus early embryos, we tested the effect of co-injection of spermine and spermidine on the functioning of exogenously microinjected in vitro-synthesized, Δβ-catenin mRNA which is known to induce a secondary head after being microinjected into a ventral vegetal blastomere in 8-celled Xenopus embryos. Microinjection of Δβ-catenin mRNA in fact induced a secondary axis with a secondary head, and co-injection of spermine or spermidine suppresses induction of the secondary head and secondary axis without drastic effects like induction of immediate cell death or execution of apoptosis at blastula stage. The inhibitory effects were dosage dependent, and at lower doses the inhibition was mainly on secondary head formation rather than on secondary axis formation. We performed similar experiments using GFP mRNA and confirmed that expression of GFP mRNA was also suppressed by co-injection of spermine. We mixed Δβ-catenin mRNA with different amounts of spermine and performed electrophoresis on agarose gels, with a finding that the prior mixing greatly suppressed mRNA migration. These results suggest that an excess amount of spermine as well as spermidine exerts inhibitory effects on mRNA translation, and that the inhibition may be due to direct binding of polyamines to mRNA and a reduction of negative charges on mRNA molecules that might also induce the formation of cross link-like networks among mRNAs.",
author = "Takamichi Mishina and Kota Fuchimukai and Kazuei Igarashi and Kosuke Tashiro and Koichiro Shiokawa",
year = "2012",
month = "2",
day = "1",
doi = "10.1007/s00726-011-0996-x",
language = "English",
volume = "42",
pages = "791--801",
journal = "Amino Acids",
issn = "0939-4451",
publisher = "Springer Wien",
number = "2-3",

}

TY - JOUR

T1 - Modification of secondary head-forming activity of microinjected Δβ-catenin mRNA by co-injected spermine and spermidine in Xenopus early embryos

AU - Mishina, Takamichi

AU - Fuchimukai, Kota

AU - Igarashi, Kazuei

AU - Tashiro, Kosuke

AU - Shiokawa, Koichiro

PY - 2012/2/1

Y1 - 2012/2/1

N2 - Polyamines are essential for cell growth and differentiation. In Xenopus early embryos, per embryo level of spermine is extremely low as compared with that of spermidine. To disclose the possible function of polyamines in Xenopus early embryos, we tested the effect of co-injection of spermine and spermidine on the functioning of exogenously microinjected in vitro-synthesized, Δβ-catenin mRNA which is known to induce a secondary head after being microinjected into a ventral vegetal blastomere in 8-celled Xenopus embryos. Microinjection of Δβ-catenin mRNA in fact induced a secondary axis with a secondary head, and co-injection of spermine or spermidine suppresses induction of the secondary head and secondary axis without drastic effects like induction of immediate cell death or execution of apoptosis at blastula stage. The inhibitory effects were dosage dependent, and at lower doses the inhibition was mainly on secondary head formation rather than on secondary axis formation. We performed similar experiments using GFP mRNA and confirmed that expression of GFP mRNA was also suppressed by co-injection of spermine. We mixed Δβ-catenin mRNA with different amounts of spermine and performed electrophoresis on agarose gels, with a finding that the prior mixing greatly suppressed mRNA migration. These results suggest that an excess amount of spermine as well as spermidine exerts inhibitory effects on mRNA translation, and that the inhibition may be due to direct binding of polyamines to mRNA and a reduction of negative charges on mRNA molecules that might also induce the formation of cross link-like networks among mRNAs.

AB - Polyamines are essential for cell growth and differentiation. In Xenopus early embryos, per embryo level of spermine is extremely low as compared with that of spermidine. To disclose the possible function of polyamines in Xenopus early embryos, we tested the effect of co-injection of spermine and spermidine on the functioning of exogenously microinjected in vitro-synthesized, Δβ-catenin mRNA which is known to induce a secondary head after being microinjected into a ventral vegetal blastomere in 8-celled Xenopus embryos. Microinjection of Δβ-catenin mRNA in fact induced a secondary axis with a secondary head, and co-injection of spermine or spermidine suppresses induction of the secondary head and secondary axis without drastic effects like induction of immediate cell death or execution of apoptosis at blastula stage. The inhibitory effects were dosage dependent, and at lower doses the inhibition was mainly on secondary head formation rather than on secondary axis formation. We performed similar experiments using GFP mRNA and confirmed that expression of GFP mRNA was also suppressed by co-injection of spermine. We mixed Δβ-catenin mRNA with different amounts of spermine and performed electrophoresis on agarose gels, with a finding that the prior mixing greatly suppressed mRNA migration. These results suggest that an excess amount of spermine as well as spermidine exerts inhibitory effects on mRNA translation, and that the inhibition may be due to direct binding of polyamines to mRNA and a reduction of negative charges on mRNA molecules that might also induce the formation of cross link-like networks among mRNAs.

UR - http://www.scopus.com/inward/record.url?scp=84861671115&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84861671115&partnerID=8YFLogxK

U2 - 10.1007/s00726-011-0996-x

DO - 10.1007/s00726-011-0996-x

M3 - Article

C2 - 21830116

AN - SCOPUS:84861671115

VL - 42

SP - 791

EP - 801

JO - Amino Acids

JF - Amino Acids

SN - 0939-4451

IS - 2-3

ER -