Immuno-histochemical localization of cyclic nucleotides in the periodontium: mechanically-stressed cells in vivo

Abstract

Cyclic AMP and cyclic GMP, considered to be cell proliferation regulators, have been reported to fluctuate in proliferating fibroblasts in vitro. The objectives of this experiment were to study the localization, distribution and staining patterns of these cyclic nucleotides in mechanically-stressed, proliferating periodontal fibroblasts in vivo. Cat canines were tipped by force applications for 0 to 48 hours and serial sagittal sections of fresh frozen, unfixed, undecalcified jaws were processed immuno-histochemically for the localization of cAMP and cGMP. Periodontal tension sites were studied microscopically. Fibroblastic staining for cAMP, which was localized mainly in the cell periphery, did not change appreciably as a result of tension. However, staining intensity increased one hour after the application of force, decreased after 6 hours and increased again at 24 hours. Staining for cGMP, initially covering the entire cell area, was concentrated over nuclei three hours after onset of tension, and diffused over cell periphery and cytoplasm thereafter. Intensity of staining for cGMP was maximal at 3 hours and low at 12 and 48 hours. These results demonstrate that stress-induced fibroblastic responses in vivo involve alterations in staining intensity for both cyclic nucleotides which may correspond with fluctuations of these regulators, reported to occur in vitro in various stages of the mitotic cycle. Moreover, this technique enables the identification of the involved cells in a stimulated, non-synchronized cell population.