Summary
First molar teeth of developing Swiss albino mice were studied for alkaline phosphatase activity. Specimens ranging from 19 days in utero to 8 days postnatal were used (prebell through appositional and mineralization stages).
The odontogenic tissues were fixed in precooled glyoxal or glutaraldehyde or formaldehyde-glutaraldehyde solutions. After 1–4 hours, the specimens were washed in buffered sucrose solutions. Both frozen and nonfrozen fixed sections were incubated for enzymic activity employing either a metal salt or an azo dye technic. Light microscopic sections were used as a basis of evaluating the fine structure observations.
The ocurrence of enzyme in the developing tooth germ was observed first in the late cap stage where it was restricted to the plasma membranes of the cells of the stratum intermedium and to a lesser degree to those of the stellate reticulum. The activity of the secretory ameloblasts was less than that of the reduced ameloblasts which were very active. In pre-odontoblasts and definitive odontoblasts activity was regularly present on the plasma membrane.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Barka, T., Anderson, P. J.: Histochemistry, theory, practice and bibliography, p. 215. New York: Harper & Row 1965.
Bradfield, J. R. G.: In: Proo. 6th Internat. Cong. Exper. Cytology. Stockholm, Suppl. 1, p. 338, 1947.
Bradfield, J. R. G.: The localization of enzymes in cells. Biol. Rev. 25, 113–157 (1950).
Burstone, M. S.: Hydrolytic enzymes in dentinogenesis and osteogenesis. In: Calcification in biological systems. Publ. No 64, p. 217, Amer. Ass. Advanc. Sci. Washington, D.C. 1960a.
Burstone, M. S.: Histochemical observations in enzymatic processes in bones and teeth. Ann. N.Y. Acad. Sci. 85, 431–444 (1960b).
Cartier, P., Picard, J.: La mineralisation du cartilage ossifiable. III. Le mecanisme de la reaction atpasique du cartilage. Bull. Soc. Chim. biol. (Paris) 37, 1159–1168 (1955).
Dalcq, A. M., Mulnard, J.: Localisation de la phosphatase alcaline dans les ebauches dentaires des rongeurs. C. R. Soc. Biol. (Paris) 147, 2040–2042 (1953).
Gomori, G.: Microtechnical demonstration of phosphatase in tissue sections. Proc. Soc. exp. Biol. (N.Y.) 42, 23–26 (1939).
Gutman, A. B., Yu, T. F.: A concept of the role or enzymes in endochondral calcification. Josiah Macy, Jr., Foundation Tr. First Conf. on Metab. Interrelations, p. 167–190 (1950).
Hugon, F., Borgers, M.: A direct lead method for the electron microscopic visualization of alkaline phosphatase activity. J. Histochem. Cytochem. 14, 429–431 (1966).
Jeener, R.: Cytochemical effects of oestradiol. Nature (Lond.) 159, 578 (1947).
Karnovsky, M. J.: A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J. Cell Biol. 27, 137A (abstr.) (1965).
Kiguel, E.: Fibrillar elements forming dentin and mineral seeding or crystal nucleation. Anat, Rec. 151, 267–274 (1965).
Kroon, D. B.: Phosphatase and the formation of protein-carbohydrate complexes. Acta anat. (Basel) 15, 317–328 (1952).
Kuftinec, M. M., Miller, S. A.: Alkaline and acid phosphatase during growth of long bones and mandibles. Calcif. Tiss. Res. 9, 173–178 (1972).
Kuhlman, R. E.: Phosphatase in epiphyseal cartilage. J. Bone Jt Surg. A 47, 545–550 (1965).
Kurahashi, Y., Yoshiki, S.: Ultrastructural localization of non-specific alkaline phosphatase activity in early stages of developing rat teeth. J. dent. Res. 49, 667 (1970).
Lennox, D. W., Provenza, D. V.: Mucopolysaccharides in odontogenesis. A histochemical and autoradiographic study. Histochemie 23, 328–341 (1970).
Leonard, E. P., Provenza, D. V.: Histochemical observations on the phosphatase activity of the developing dental primordia in the Swiss albino mouse as detected by azo dye and osmium-capture methods. Arch. oral Biol. 15, 635–643 (1970).
Leonard, E. P., Provenza, D. V.: Comparative study of the various methods for the ultrastructural localization of alkaline phosphatase activity. Histochemie 30, 1–12 (1972).
Luft, J.: Improvements in epoxy resin embedding methods. J. biophys. biochem. Cytol. 9, 409–414 (1961).
Mizutani, A., Barrnett, R. J.: Fine structural demonstration of phosphatase activity at pH 9. Nature (Lond.) 206, 1001–1003 (1965).
Pritchard, J. J.: A cytological and histochemical study of bone and cartilage formation in the rat. J. Anat. (Lond.) 86, 259–277 (1952).
Reith, E. J., Butcher, E. O.: Microanatomy and histochemistry of amelogenesis. In: Structural and chemical organization of teeth, ed. A. E. W. Miles, vol. I. New York: Academic Press 1967.
Reynolds, E. S.: The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J. Cell Biol. 17, 208–212 (1963).
Robison, R.: The possible significance of hexosephosphoric esters in ossification. Biochem. J. 1, 286–293 (1923).
Sabatini, D. D., Bensch, K., Barrnett, R. J.: The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J. Cell Biol. 17, 19–58 (1963).
Seligman, A. M., Kawashima, T., Ueno, H., Katzoff, L., Hanker, J. S.: Histochemical demonstration of acid and alakline phosphatase using 2-naphthylthiolphosphate as substrate and osmiophilic derivatives as end products. Acta histochem. cytochem. 3, 29–40 (1970).
Siffert, R. S.: The role of alkaline phosphatase in osteogenesis. J. exp. Med. 93, 415–426 (1951).
Sisca, R. F., Provenza, D. V.: Initial dentin formation in human deciduous teeth. An electron miscroscope study. Calcif. Tiss. Res. 9, 1–16 (1972).
Sisca, R. F., Provenza, D. V., Narbaitz, R.: The submicroscopic localization of alkaline phosphatase in the stratum intermedium of the Macaca, mulatta tooth germ in the appoitional stage. Internat. Ass. for Dent. Res. Preprinted abstracts, 46th General Meeting, Abstract 617 (1968).
Ten Cate, A. R.: The distribution of alkaline phosphatase in the human tooth germ. Arch. oral Biol. 7, 195–205 (1962).
Ten Cate, A. R.: Alkaline phosphatase activity and the formation of human circumpulpal dentine. Arch. oral Biol. 11, 267–268 (1966).
Thilander, H.: Localization of alkaline phosphatase in the tooth germs. of albino rats. Acta odont. scand. 17, 529–538 (1959).
Vaes, G., Jacques, P.: Study on bone enzyme. The assay of acid hydrolyses and other enzymes in bone tissue. Biochem. J. 97, 380–388 (1968).
Watson, M. L.: Staining of tissue sections for electron microscopy with heavy metal. J. biophys. biochem. Cytol. 4, 475–478 (1958).
Wergedal, J. E.: Enzymes of protein and phosphate catabolism in rat bone. I. Enzyme property in normal rat. II. Changes in vitamin D deficiency. Calcif. Tiss. Res. 3, 55–73 (1969).
Woltgens, J. H. M., Bonting, S. L., Bijvoet, O. L. M.: Relationships between alkaline phosphatase and inorganic pyrophosphatase in young hamster molars. Calcif. Tiss. Res. 4, Suppl., 39 (1970).
Yoshiki, S., Kurahashi, Y.: A light and electron microscopic study of alkaline phosphatase activity in the early stage of dentinogenesis in the young rat. Arch. oral Biol. 16, 1143–1154 (1971).
Author information
Authors and Affiliations
Additional information
This study was supported by USPHS Research Grant # DE 2800-02, National Institutes of Health.
Rights and permissions
About this article
Cite this article
Leonard, E.P., Provenza, D.V. Alkaline phosphatase activity in sequential mouse molar tooth development an electron microscopic study. Histochemie 34, 343–354 (1973). https://doi.org/10.1007/BF00306306
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00306306