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Characterization of transgenic mice with overexpression of spermidine synthase

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Abstract

A composite cytomegalovirus-immediate early gene enhancer/chicken β-actin promoter (CAG) was utilized to generate transgenic mice that overexpress human spermidine synthase (SpdS) to determine the impact of elevated spermidine synthase activity on murine development and physiology. CAG-SpdS mice were viable and fertile and tissue SpdS activity was increased up to ninefold. This increased SpdS activity did not result in a dramatic elevation of spermidine or spermine levels but did lead to a 1.5- to 2-fold reduction in tissue spermine:spermidine ratio in heart, muscle and liver tissues with the highest levels of SpdS activity. This new mouse model enabled simultaneous overexpression of SpdS and other polyamine biosynthetic enzymes by combining transgenic animals. The combined overexpression of both SpdS and spermine synthase (SpmS) in CAG-SpdS/CAG-SpmS bitransgenic mice did not impair viability or lead to overt developmental abnormalities but instead normalized the elevated tissue spermine:spermidine ratios of CAG-SpmS mice. The CAG-SpdS mice were bred to MHC-AdoMetDC mice with a >100-fold increase in cardiac S-adenosylmethionine decarboxylase (AdoMetDC) activity to determine if elevated dcAdoMet would facilitate greater spermidine accumulation in mice with SpdS overexpression. CAG-SpdS/MHC-AdoMetDC bitransgenic animals were produced at the expected frequency and exhibited cardiac polyamine levels comparable to MHC-AdoMetDC littermates. Taken together these results indicate that SpdS levels are not rate limiting in vivo for polyamine biosynthesis and are unlikely to exert significant regulatory effects on cellular polyamine content and function.

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Abbreviations

AdoMet:

S-Adenosylmethionine

AdoMetDC:

S-Adenosylmethionine decarboxylase

AZ:

Antizyme

CAG:

Composite cytomegalovirus immediate early gene enhancer-chicken β-actin promoter

dcAdoMet:

Decarboxylated S-adenosylmethionine

DFMO:

α-Difluoromethylornithine

Gy:

Gyro

HA:

Hemagglutinin

MHC:

α-Myosin heavy chain

ODC:

Ornithine decarboxylase

PCR:

Polymerase chain reaction

SpdS:

Spermidine synthase

SpmS:

Spermine synthase

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Acknowledgments

The authors wish to thank Tom Salada of the Penn State University Transgenic Mouse Facility for microinjection of the CAG-SpdS construct, the Penn State University College of Medicine Macromolecular Synthesis and DNA Sequencing Cores, and the technicians of the Penn State University College of Medicine Department of Comparative Medicine for expert animal care. We thank Dr. J.A. Sawicki for providing the pCX-EGFP plasmid and Dr. M. Okabe for permission to use this construct in our experiments. Supported by National Institutes of Health grants CA-018138 (DJF) and GM-26290 (AEP).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, PO Box 850, 500 University Drive, Hershey, PA, 17033, USA

    Chenxu Shi, Patricia A. Welsh, Suzanne Sass-Kuhn, Xiaojing Wang, Anthony E. Pegg & David J. Feith

  2. Department of Pharmacology, Pennsylvania State University College of Medicine, PO Box 850, 500 University Drive, Hershey, PA, 17033, USA

    Diane E. McCloskey & Anthony E. Pegg

  3. Department of Cellular and Molecular Physiology (H166), Pennsylvania State University College of Medicine, Room C4740A, 500 University Drive, Hershey, PA, 17033, USA

    David J. Feith

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  1. Chenxu Shi
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  2. Patricia A. Welsh
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  3. Suzanne Sass-Kuhn
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  4. Xiaojing Wang
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  5. Diane E. McCloskey
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  6. Anthony E. Pegg
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  7. David J. Feith
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Correspondence to David J. Feith.

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Shi, C., Welsh, P.A., Sass-Kuhn, S. et al. Characterization of transgenic mice with overexpression of spermidine synthase. Amino Acids 42, 495–505 (2012). https://doi.org/10.1007/s00726-011-1028-6

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  • DOI: https://doi.org/10.1007/s00726-011-1028-6

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