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miR-212 and miR-132 are required for epithelial stromal interactions necessary for mouse mammary gland development

Nature Genetics volume 42, pages 1101–1108 (2010)Cite this article

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Abstract

MicroRNAs are small noncoding RNAs that carry out post-transcriptional regulation of the expression of their target genes. However, their roles in mammalian organogenesis are only beginning to be understood. Here we show that the microRNA-212/132 family (which comprises miR-212 and miR-132) is indispensable during the development of the mammary glands in mice, particulary for the regulation of the outgrowth of the epithelial ducts. Mammary transplantation experiments revealed that the function of the miR-212/132 family is required in the stroma but not in the epithelia. Both miR-212 and miR-132 are expressed exclusively in mammary stroma and directly target the matrix metalloproteinase MMP-9. In glands that lack miR-212 and miR-132, MMP-9 expression increases and accumulates around the ducts. This may interfere with collagen deposition and lead to hyperactivation of the tumor growth factor-β signaling pathway, thereby impairing ductal outgrowth. Our results identify the miR-212/132 family as one of the main regulators of the epithelial-stromal interactions that are required for proper pubertal development of the mammary gland.

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Figure 1: Genetic deletion of miR-212/132 in mice results in mammary gland defects associated with impaired nourishment of pups.
Figure 2: miR-212/132−/− mammary glands have a defect in pubertal ductal outgrowth, but not in ductal side branching and lobuloalveolar differentiation.
Figure 3: miR-212/132−/− mammary epithelia have normal ductal architecture.
Figure 4: miR-212/132−/− mammary epithelia have normal capacity for ductal outgrowth.
Figure 5: The miR-212/132−/− stroma is not permissive for pubertal outgrowth of the epithelial ducts.
Figure 6: In the mammary gland, miR-212 and miR-132 are exclusively expressed in the stromal cells, but not in the epithelia.
Figure 7: Loss of miR-212 and miR-132 causes high MMP-9 levels and hyperactivity of TGF-β pathway in mutant mammary glands.

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Acknowledgements

We thank S. Geisendorf for technical assistance, S. Hille for DNA sequencing, S. Mahsur and U. Franke for technical help in generation of knockout mouse line, A. Kurth, A. Driehorst, K. Kiel and U. Teichman for animal care work, M. Smalley for discussion about mammary epithelial transplantation method, N. Ba Tiep for technical help with collecting blood samples, A. Yalcin for creative contribution to the initial phase of the work, and P. Gruss and G. Eichele for encouragement and support. Funded by the Max Planck Society. Additional funding was provided by the Deutsche Forschungsgemeinschaft (DFG, GR 536/9-2 and GR 536/11-1 to B.G. and TH 903/7-2 to T.T.).

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  1. Ahmet Ucar

    Present address: Present address: Cellular Senescence Group, German Cancer Research Center (DKFZ), Heidelberg, Germany.,

Authors and Affiliations

  1. Department of Molecular Cell Biology, Max Planck Institute of Biophysical Chemistry, Goettingen, Germany

    Ahmet Ucar & Kamal Chowdhury

  2. Georg-Speyer-Haus, Institute for Biomedical Research, Frankfurt am Main, Germany

    Vida Vafaizadeh, Petra A B Klemmt & Bernd Groner

  3. Department of Endocrinology, University of Goettingen, Goettingen, Germany

    Hubertus Jarry

  4. Institute for Molecular and Translational Therapeutic Strategies, Hannover Medical School, Germany

    Jan Fiedler & Thomas Thum

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Contributions

A.U. and K.C. developed the concept of this study. A.U. performed all experiments except the mammary epithelial transplantation analyses, which were performed by V.V. and P.A.B.K. in the laboratory of B.G., serum hormone level analyses, which were done by H.J., and luciferase assays, which were done by J.F. in the laboratory of T.T. The manuscript was written by A.U. and K.C. Important suggestions were made by B.G., V.V. and T.T. which improved the quality of the manuscript. All authors contributed to the discussion of the data and commented on the final version of the manuscript.

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Correspondence to Kamal Chowdhury.

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The authors declare no competing financial interests.

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Ucar, A., Vafaizadeh, V., Jarry, H. et al. miR-212 and miR-132 are required for epithelial stromal interactions necessary for mouse mammary gland development. Nat Genet 42, 1101–1108 (2010). https://doi.org/10.1038/ng.709

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