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Development of transgenic mice overexpressing mouse carbonyl reductase 1

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Abstract

Background

Carbonyl reductase 1 (CBR1) is a nicotinamide adenine dinucleotide phosphate (NADPH)-dependent reductase with broad substrate specificity. CBR1 catalyzes the reduction of numerous carbonyl compounds, including quinones, prostaglandins, menadione, and multiple xenobiotics, while also participating in various cellular processes, such as carcinogenesis, apoptosis, signal transduction, and drug resistance. In this study, we aimed to generate transgenic mice overexpressing mouse Cbr1 (mCbr1), characterize the mCbr1 expression in different organs, and identify changes in protein expression patterns.

Methods and Results

To facilitate a deeper understanding of the functions of CBR1, we generated transgenic mice overexpressing CBR1 throughout the body. These transgenic mice overexpress 3xFLAG-tagged mCbr1 (3xFLAG-mCbr1) under the CAG promoter. Two lines of transgenic mice were generated, one with 3xFLAG-mCbr1 expression in multiple tissues, and the other, with specific expression of 3xFLAG-mCbr1 in the heart. Pathway and network analysis using transgenic mouse hearts identified 73 proteins with levels of expression correlating with mCbr1 overexpression. The expression of voltage-gated anion channels, which may be directly related to calcium ion-related myocardial contraction, was also upregulated.

Conclusion

mCbr1 transgenic mice may be useful for further in vivo analyses of the molecular mechanisms regulated by Cbr1; such analyses will provide a better understanding of its effects on carcinogenesis and cardiotoxicity of certain cancer drugs.

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Data Availability

NGS analysis data are available online using the accession number “DRA012939” for DDBJ/EMBL/GenBank.The proteomic data are available online using the accession number “PXD027170” for ProteomeXchange [42], and the accession number “JPST001246” for the jPOST repository [43].

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Acknowledgements

The authors thank Hiroshi Kijima for his assistance with the immunohistochemistry analysis; Takeshi Shimizu, Shoko Nagata, and Hirotaka Fujita for their technical assistance; and Miyu Miyazaki for her technical support in the mass spectrometry analysis.

Author information

Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Graduate School of Medicine, Hirosaki University, 5 Zaifu-cho, 036-8562, Hirosaki, Aomori, Japan

    Minako Yokoyama, Yuka Kadonosawa & Yoshihito Yokoayama

  2. Department of Biochemistry and Genome Biology, Graduate School of Medicine, Hirosaki University, 5 Zaifu-cho, 036-8562, Hirosaki, Aomori, Japan

    Toshitsugu Fujita & Hodaka Fujii

  3. Department of Stress Response Science, Center for Advanced Medical Research, Graduate School of Medicine, Hirosaki University, 5 Zaifu-cho, 036-8562, Hirosaki, Aomori, Japan

    Yota Tatara

  4. Genome Information Research Center, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, 565-0871, Suita, Osaka, Japan

    Daisuke Motooka & Masahito Ikawa

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  1. Minako Yokoyama
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  2. Toshitsugu Fujita
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Contributions

M.Y., H.F., and Y.Y. conceived the project; T.F. and H.F. designed and supervised the project; Y.T. performed the LC-MS/MS analysis; D.M. performed the NGS analysis; M.I. generated the transgenic mice; M.Y. and Y.K. performed the other experiments; and M.Y., T.F., H.F., and Y.Y. wrote the manuscript. All authors have read and approved the final version of the manuscript.

Corresponding author

Correspondence to Yoshihito Yokoayama.

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Funding

None.

Competing Interests:

Y.Y. and M.Y. have filed a patent application for the transgenic mice with details as follows: Name: Transgenic non-human animals; Number: Japanese Patent Application No. 2022-009381; Status: Under review; Specific aspect of manuscript covered in the patent application: All the information reported in this study.

Ethics approval:

All animal experiments were approved by the Institutional Animal Care and Use Committee at the Research Institute for Microbial Diseases, Osaka University (4111) and Hirosaki University (M19024). This study complied with the ARRIVE guidelines. All methods were performed in accordance with the relevant guidelines and regulations.

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Yokoyama, M., Fujita, T., Kadonosawa, Y. et al. Development of transgenic mice overexpressing mouse carbonyl reductase 1. Mol Biol Rep 50, 531–540 (2023). https://doi.org/10.1007/s11033-022-07994-x

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