Confocal laser scanning and microscopic study of tartrate-resistant acid phosphatase-positive cells in the dental follicle during early morphogenesis of mouse embryonic molar teeth

Abstract

Tartrate-resistant acid phosphatase (TRAPase)-positive cells were examined during the early morphogenesis of mouse mandibular first molar teeth, using a simultaneous azo dye-coupling technique and confocal laser scanning microscopy (CLSM). CLSM of paraffin-embedded sections revealed that cells exhibiting TRAPase fluorescence appeared along the marginal region of the condensed dental mesenchyme, not yet the dental follicle, at embryonic day 12 (E12). The follicle was recognized histologically as a population of fibroblastic cells in tooth organs at E14, and TRAPase-positive cells were localized among the follicular cells. Light-microscopic observations of microsliced thick sections showed the overall morphology of the TRAPase-positive cells. A few positive cells of E13 tooth organs were oval, and the intense reaction products of TRAPase revealed complex processes on positive cells in E14 tooth organs. Three-dimensional fluorescent images of TRAPase-positive cells using microsliced sections and CLSM showed their unique morphology of long processes with small spine-like projections at E14. Thus, TRAPase-positive cells appeared in the region of the prospective follicle at the bud stage (E12 and 13) before the formation of the follicle proper. In addition, the formation of the follicle and the appearance of TRAPase-positive cells with unique morphology were correlated in their developmental stage (E14). The findings suggest that the TRAPase-positive cells may be involved in the formation of the dental follicle in early tooth morphogenesis.