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Journal of Histochemistry and Cytochemistry, Vol. 46, 459-468, April 1998, Copyright © 1998, The Histochemical Society, Inc.

ARTICLE

Pex mRNA Is Localized in Developing Mouse Osteoblasts and Odontoblasts

Andréa Frota Ruchona,d, Mieczyslaw Marcinkiewiczb, Géraldine Siegfrieda, Harriet S. Tenenhousec, Luc DesGroseillersa, Philippe Crinea, and Guy Boileaua
a Département de Biochimie, Institut de Recherches Cliniques de Montréal et Département de Médecine, Université de Montréal, Montréal, Canada
b Laboratoire de Neuroendocrinologie Moléculaire, Institut de Recherches Cliniques de Montréal et Département de Médecine, Université de Montréal, Montréal, Canada
c Department of Pediatrics, McGill University–Montreal Children's Hospital Research Institute and Department of Human Genetics, McGill University, Montreal, Canada
d Departamento de Morfologia, Universidade Federal do Ceará, Fortaleza, Brasil

Correspondence to: Guy Boileau, Département de Biochimie, Université de Montréal, CP 6128, Succ. Centre-Ville, Montréal, Qc, Canada H3C 3J7.

Mutations in PEX, a phosphate-regulating gene with homology to endopeptidase on the X chromosome, were recently identified in patients with X-linked hypophosphatemia (XLH), an inherited disorder of phosphate homeostasis characterized by growth retardation and rachitic and osteomalacic bone disease. To understand the mechanism by which loss of PEX function elicits the mutant phenotype, a study of its mRNA localization and ontogenesis was undertaken. Using the reverse transcriptase–nested polymerase chain reaction (RT-nested PCR) with polyA+ RNA purified from mouse testis, a 337-BP Pex cDNA fragment was generated and cloned in the pCRII plasmid. The cDNA was used to generate sense and anti-sense Pex riboprobes for in situ hybridization (ISH) and Northern analysis. To survey a large number of different tissues, sagittal sections of embryos and newborn mice were examined. ISH showed the presence of Pex mRNA in osteoblasts and odontoblasts. Pex gene expression was detectable on Day 15 of embryonic development, which coincides with the beginning of intercellular matrix deposition in bones. Finally, Northern analysis of total RNA from calvariae and teeth of 3-day-old and adult mice showed that the abundance of the 7-KB Pex transcript is decreased in adult bones and in nongrowing teeth. The present study demonstrates that Pex mRNA is expressed in bones and teeth and suggests that this putative endopeptidase plays an important role in the development of these tissues. (J Histochem Cytochem 46:459–468, 1998)

Key Words: bone, teeth, extracellular matrix, ontogenesis, X-linked hypophosphatemia, in situ hybridization, Northern blot analysis


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