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Microarray Profiling of Diaphyseal Bone of Rats Suffering from Hypervitaminosis A

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Abstract

Vitamin A is the only known compound that produces spontaneous fractures in rats. In an effort to resolve the molecular mechanism behind this effect, we fed young male rats high doses of vitamin A and performed microarray analysis of diaphyseal bone with and without marrow after 1 week, i.e., just before the first fractures appeared. Of the differentially expressed genes in cortical bone, including marrow, 98% were upregulated. In contrast, hypervitaminotic cortical bone without marrow showed reduced expression of 37% of differentially expressed genes. Gene ontology (GO) analysis revealed that only samples containing bone marrow were associated with a GO term, which principally represented extracellular matrix. This is consistent with the histological findings of increased endosteal/marrow osteoblast number. Fourteen genes, including Cyp26b1, which is known to be upregulated by vitamin A, were selected and verified by real-time PCR. In addition, immunohistochemical staining of bone sections confirmed that the bone-specific molecule osteoadherin was upregulated. Further analysis of the major gene-expression changes revealed apparent augmented Wnt signaling in the sample containing bone marrow but reduced Wnt signaling in cortical bone. Moreover, induced expression of hypoxia-associated genes was found only in samples containing bone marrow. Together, these results highlight the importance of compartment-specific analysis of bone and corroborate previous observations of compartment-specific effects of vitamin A, with reduced activity in cortical bone but increased activity in the endosteal/marrow compartment. We specifically identify potential key osteoblast-, Wnt signaling-, and hypoxia-associated genes in the processes leading to spontaneous fractures.

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Acknowledgment

We thank Uppsala Array Platform for their technical assistance and Valeria Giandomenico for critical reading of the manuscript. This work was supported by the Swedish Society of Medicine (T. L.) and the Swedish Medical Research Council (H. M., G. A.).

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Authors and Affiliations

  1. Department of Medical Sciences, Section of Clinical Pharmacology, Uppsala University, UAS, Ing 70 3tr Foa2 Lab22, 75185, Uppsala, Sweden

    Thomas Lind, Lijuan Hu, Annica Jacobson & Håkan Melhus

  2. Department of Medical Sciences, Section of Occupational and Environmental Medicine, Uppsala University, 75185, Uppsala, Sweden

    P. Monica Lind

  3. Oral Biology, Department of Dental Medicine, Karolinska Institutet, P.O. Box 4064, 14104, Huddinge, Sweden

    Rachael Sugars

  4. Division of Pathology, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital, 14152, Huddinge, Sweden

    Göran Andersson

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  1. Thomas Lind
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Correspondence to Thomas Lind.

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Lind, T., Hu, L., Lind, P.M. et al. Microarray Profiling of Diaphyseal Bone of Rats Suffering from Hypervitaminosis A. Calcif Tissue Int 90, 219–229 (2012). https://doi.org/10.1007/s00223-011-9561-6

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  • DOI: https://doi.org/10.1007/s00223-011-9561-6

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