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

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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, ∆β-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 ∆β-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 ∆β-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.

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Abbreviations

MBT:

Midblastula transition

SAMDC:

S-adenosylmethionine decarboxylase

ODC:

Ornithine decarboxylase

SAM:

S-adenosylmethionine

GFP mRNA:

Green fluorescent protein mRNA

FITC-dextran:

Fluorescein isothiocyanate-dextran

PCR:

Polymerase chain reaction

MBS:

Modified Barth’s solution

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Acknowledgments

The authors thank Drs. Senji Takahashi and Masayuki Kajitani, Department of Biosciences, Teikyo University, for their supports in our research. Thanks are also due to Professor Akira Komatsu, Department of Judo Therapy, Teikyo University for trial to apply statistical treatment (t test) for the data obtained. The present work was supported by 2010 Okinaga Special Reseach Support Fund in Utsunomiya Campus in Teikyo University, Utsunomiya, Japan.

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Authors and Affiliations

  1. Department of Biosciences, School of Science and Engineering, Teikyo University, 1-1 Toyosatodai, Utsunomiya, 320-8551, Tochigi, Japan

    Takamichi Mishina, Kota Fuchimukai & Koichiro Shiokawa

  2. Department of Judo Therapy, Faculty of Medical Technology, Teikyo University, 1-1 Toyosatodai, Utsunomiya, 320-8551, Tochigi, Japan

    Koichiro Shiokawa

  3. Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana Chuo-ku, Chiba, 260-8675, Japan

    Kazuei Igarashi

  4. Molecular Gene Technics, Genetic Resources Technology, Kyushu University, Hakozaki 6-10-1, Fukuoka, 812-8581, Japan

    Kosuke Tashiro

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  1. Takamichi Mishina
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  2. Kota Fuchimukai
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  3. Kazuei Igarashi
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Correspondence to Koichiro Shiokawa.

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Mishina, T., Fuchimukai, K., Igarashi, K. et al. Modification of secondary head-forming activity of microinjected ∆β-catenin mRNA by co-injected spermine and spermidine in Xenopus early embryos. Amino Acids 42, 791–801 (2012). https://doi.org/10.1007/s00726-011-0996-x

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  • DOI: https://doi.org/10.1007/s00726-011-0996-x

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