Nmunolocalización de factores de crecimiento de fibroblastos ácido y básico mientras el ratón está odontogénesis

Int..I.

De\'. Bioi. 36: 381-389 (1991)

381

Original

Article

Immunolocalization of acidic and basic fibroblast growth factors during mouse odontogenesis
YVES CAM", MARIE-ROSE NEUMANN" LISA OLIVER', DANIEL RAULAIS', and JEAN-VICTOR RUCH' THIERRY JANET3#
7/nstitut de 8iologie Medica/e, INSERM CJF 88-08, Universite Paris and 3Laboratoire de Neurobiologie Ontogenique Louis Pasteur,Faculte de Medecine, Strasbourg, 21NSERM Unite 118, CNRS UPR 417 - Centre de Neurochimie, Strasbourg, France

Acidic and basic fibroblast growth factors laFGF and bFGFI, are both known to bind to extracellular matrix components, particularly proteoheparan sulfates, and to regulate in vitro proliferation, differentiation and morphology of cells of neuroectodermal and mesodermal origins. Theirpatterns of distribution were studied during mouse odontogenesis by means of indirect immunofluorescence and immunoperoxidase histochemistry on frozen fixed sections and after Bouin's fixative and paraffin embedding. Localization of aFGF on frozen fixed sections was observed in the oral epithelium, dental lamina and oral mesenchyme (day-12 of gestation), the stellate reticulum and oral epithelium(day-14), the stratum intermedium and atthe basal and apical poles ofpreameloblasts at bell stage. After birth aFGF epitopes were localized within the predentin-dentin area, the stratum intermedium and at the secretory pole of ameloblasts. There was no staining with anti-aFGF antibodies after Bouin's fixative and paraffin embedding. In contrast, using this protocol, intense stainings were foundwith anti-bFGF antibodies predominantly within dental and peridental basement membranes and mesenchyme: staining of the dental basement membranes was transient (bud and cap stage) and discontinuous; a preferential concentration of bFGF epitopes in the condensed dental mesenchyme of incisors (cap stage) and the dental papillae mesenchymal cells of molars (bell stage) was observed in the posterior andthe cervical part of tooth germs. An intense immunostaining of the stellate reticulum with anti-bFGF antibodies was also found on paraffin sections from bud to bell stage. Localization ofbFGF on frozen fixed sections was observed in the dental lamina, stellate reticulum and dental basement membranes at bud and cap stage, the stratum intermedium and atthe secretory pole of ameloblasts afterbirth. Treatment of sections with NaCI (2-3M) solutions and heparitinase diminished, but did not abolish, specific immunostaining obtained with bFGF antibodies. Our results suggest that, among growth factors, a- and bFGFs might intervene in different ways during odontogenesis, particularly through binding to the cellular and/or extracellular matrix heparan sulfate-containing molecules, and mayparticipate in the control of proliferation, determination and terminal differentiation of preodontoblasts and preameloblasts.
KEY WORDS: lis.\'/lf fi.'\at;on imllllfr/ohistochl'lIIislry, (u'idir and b(Hirfihroh/ast growthfrU'lors, mOIlSf' or!olll0K"lIf'sis,

ABSTRACT

Introduction

1988; Thesleff et al., 1990; Lesot et al., 1990; Mark et al., 1990;
Kronmiller

et al., 1991a, b; Vainio etal..1991; Hall and Ekanayake,

Odontogenesis is a well studied example ofcel! kinetic-dependent developmental processes that are mediated through homotypic and heterotypic cell interactions. Most of these interactions involve the temporal and spatial action of extracellular matrices first as solid substrata that interact with plasma membrane receptors and ligands and second as potential reservoirsfor diffusible molecules such as growth!actors (Camef al.,1987; Ruch,1987, 1990; Slavkin,
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