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Journal of Histochemistry and Cytochemistry, Vol. 47, 1541-1552, December 1999, Copyright © 1999, The Histochemical Society, Inc.

ARTICLE

Differential Expression and Activity of Tissue-nonspecific Alkaline Phosphatase (TNAP) in Rat Odontogenic Cells In Vivo

Dominique Hottona,b, Nicole Mauroa, Frédéric Lézota, Nadine Foresta, and Ariane Berdala
a Laboratoire de Biologie–Odontologie, Institut Biomédical des Cordeliers, Paris, France
b INSERM U458, Paris, France

Correspondence to: Dominique Hotton, Laboratoire de Biologie–Odontologie, EA 2380, UP7, Institut Biomédical des Cordeliers, 15–21 rue de l'Ecole de Médecine, 75006 Paris, France.

Among the four existing isoforms of alkaline phosphatase (AP), the present study is devoted to tissue-nonspecific alkaline phosphatase (TNAP) in mineralized dental tissues. Northern blot analysis and measurements of phosphohydrolase activity on microdissected epithelium and ectomesenchyme, in situ hybridization, and immunolabeling on incisors confirmed that the AP active in rodent teeth is TNAP. Whereas the developmental pattern of TNAP mRNA and protein and the previously described activity were similar in supra-ameloblastic and mesenchymal cells, they differed in enamel-secreting cells, the ameloblasts. As previously shown for other proteins involved in calcium and phosphate handling in ameloblasts, a biphasic pattern of steady-state TNAP mRNA levels was associated with additional variations in ameloblast TNAP protein levels during the cyclic modulation process. Although the association of TNAP upregulation and the initial phase of biomineralization appeared to be a basic feature of all mineralized tissues, ameloblasts (and to a lesser extent, odontoblasts) showed a second selectively prominent upregulation of TNAP mRNA/protein/activity during terminal growth of large enamel crystals only, i.e., the maturation stage. This differential expression/activity for TNAP in teeth vs bone may explain the striking dental phenotype vs bone reported in hypophosphatasia, a hereditary disorder related to TNAP mutation. (J Histochem Cytochem 47:1541–1552, 1999)

Key Words: alkaline phosphatase, bone, dentin, enamel, mineralization, vitamin D, hypophosphatasia, in situ hybridization, immunolocalization


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