Expression of type I collagen pro-alpha 2 chain mRNA in adult human permanent teeth as revealed by in situ hybridization.
The expression of the gene COL1A2, coding for the pro-alpha 2 chain of type I pro-collagen, was analyzed in fully developed human permanent teeth. The teeth were fixed with formalin, demineralized with EDTA for about ten weeks, and embedded in paraffin. Pro-alpha 2(I) mRNA was localized in the sections by in situ hybridization, with use of [35S)]-labeled single-stranded RNA probes. The amount of mRNA for pro-alpha 2(I) collagen chain, as indicated by the relative densities of silver grains and the grain counts per cell in autoradiography, was high in odontoblasts, whereas in pulpal fibroblasts it was low. High levels of pro-alpha 2(I)mRNA expression were also present in those odontoblasts which had elaborated new dentin matrix in response to dental caries. Expression in the periodontal ligament, including the cementoblast layer, was slightly stronger than that in odontoblasts. The intense expression of pro-alpha 2(I) mRNA in odontoblasts of adult teeth suggests that even after the completion of primary dentin formation, they continue to synthesize heterotrimeric type I collagen molecules. Cell type-specific differences in the expression of pro-alpha 2(I) mRNA imply that type I collagen probably plays a major role in the regulation of the structure and function of dental tissues. Finally, in situ hybridization enabled pro-alpha 2(I) collagen mRNA to be detected in tissue sections even after prolonged demineralization, and thus it proved to be a valuable technique for analysis of gene expression in adult dental tissues, as shown here for COL1A2.
Lukinmaa
P L
PL
Department of Dental Radiology/Oral Pathology, University of Helsinki, Finland.
Vaahtokari
A
A
Vainio
S
S
Thesleff
I
I
D000328:Adult; D003094:Collagen / Q000096:biosynthesis / Q000235:genetics*; D003782:Dental Pulp / Q000737:chemistry*; D003804:Dentin / Q000737:chemistry; D015870:Gene Expression; D006801:Humans; D008963:Molar / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D009804:Odontoblasts / Q000737:chemistry*; D010513:Periodontal Ligament / Q000737:chemistry*; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis*
D015347:RNA Probes; D012333:RNA, Messenger; D003094:Collagen