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MRI detection of transcriptional regulation of gene expression in transgenic mice

Nature Medicine volume 13, pages 498–503 (2007)Cite this article

Abstract

Ferritin, the iron storage protein, was recently suggested to be a candidate reporter for the detection of gene expression by magnetic resonance imaging (MRI). Here we report the generation of TET:EGFP-HAferritin (tet-hfer) transgenic mice, in which tissue-specific inducible transcriptional regulation of expression of the heavy chain of ferritin could be detected in vivo by MRI. We show organ specificity by mating the tet-hfer mice with transgenic mice expressing tetracycline transactivator (tTA) in liver hepatocytes and in vascular endothelial cells. Tetracycline-regulated overexpression of ferritin resulted in specific alterations of the transverse relaxation rate (R2) of water. Transgene-dependent changes in R2 were detectable by MRI in adult mice, and we also found fetal developmental induction of transgene expression in utero. Thus, the tet-hfer MRI reporter mice provide a new transgenic mouse platform for in vivo molecular imaging of reporter gene expression by MRI during both embryonic and adult life.

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Figure 1: Generation of tet-hfer MRI reporter mice.
Figure 2: MRI detection of endothelium-selective overexpression of h-ferritin in the adult mouse brain.
Figure 3: In utero MRI detection of VE-cadherin mediated expression in E13.5 fetal liver and heart.
Figure 4: MRI detection of h-ferritin overexpression by liver hepatocytes.
Figure 5: Liver overexpression of h-ferritin in the liver-hfer mice.

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Acknowledgements

We would like to acknowledge helpful discussions with A. Koretsky, P. Bendel, E. Keshet and Y. Dor. We would like to thank G. Damari and E. Ino for their help in generation and breeding of the transgenic mice. This work was supported by the Israel Science Foundation 391/02 and by the Minerva Foundation (to M.N.).

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Author notes

  1. Batya Cohen and Keren Ziv: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biological Regulation, Weizmann Institute, Rehovot, 76100, Israel

    Batya Cohen, Keren Ziv, Vicki Plaks, Tomer Israely & Michal Neeman

  2. Department of Veterinary Resources, Weizmann Institute, Rehovot, 76100, Israel

    Vyacheslav Kalchenko & Alon Harmelin

  3. Department of Pathology, Beth Israel Deaconess Medical Center and Harvard Medical School, 330 Brookline Ave., Boston, 02215, Massachusetts, USA

    Laura E Benjamin

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Contributions

B.C. generated the construct and derived the transgenic mice; K.Z., V.P. and M.N. performed the MRI experiments and conducted the data analyses; K.Z., V.P., B.C., T.I., V.K. and A.H. contributed to the ex vivo fluorescence and histological analysis, L.E.B. provided the VE-cadherin mice; M.N., B.C., K.Z. and V.P. wrote the manuscript and M.N. supervised the project.

Corresponding author

Correspondence to Michal Neeman.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Histological analysis of multiple organs in male VE cadherin h-fer mice. (PDF 136 kb)

Supplementary Fig. 2

Ferritin expression during embryonic development. (PDF 301 kb)

Supplementary Fig. 3

Prussian Blue staining of ferritin iron. (PDF 147 kb)

Supplementary Video 1

Contrast enhanced 3D gradient echo MRI of E13.5 pregnant mouse. The dataset was acquired at the end of the spin echo measurement of R2 relaxation, after intravenous administration of biotin-BSA-GdDTPA. Hyperintense areas show the maternal vasculature including maternal circulation in the placenta (same mouse as Figure 3). (AVI 885 kb)

Supplementary Video 2

Maximal intensity projection revealing the placenta of E13.5 pregnancy. Movie was derived from the 3D contrast enhanced gradient echo dataset of movie S3. (AVI 652 kb)

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Cohen, B., Ziv, K., Plaks, V. et al. MRI detection of transcriptional regulation of gene expression in transgenic mice. Nat Med 13, 498–503 (2007). https://doi.org/10.1038/nm1497

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