Influences of fixatives on the immunodetection of calcitonin in mouse "C" cells during pre- and post- natal development.

Abstract

The presence of calcitonin in fetal and postnatal mouse "C" cell was demonstrated by radioimmunoassays of fetal and adult thyroid extracts and by immunohistochemistry following the use of Bouin's fixative from 18 days postcoitum onwards. However, after fixation with formaldehyde no calcitonin could be detected in fetal or early postnatal samples, although it was still easily detectable in adult tissues treated with this fixative, raising the possibility that there may be age-dependent differences in the chemistry or physiological state of intracellular calcitonin. Experiments conducted with several other fixatives and the results of the radioimmunoassays suggest that the loss of calcitonin reactivity is not due to differences in the anti-genic sites of the molecule at the two stages in development. Neither can the positive detection of fetal material treated with Bouin's fixative be due to any action of this liquid. However, the inhibiting action of the aldehydes could be related to the preservation of the granule membranes in the fetal "C" cells: our ultrastructural studies show that the granules can be morphologically classified in dense-core and homogeneous matrix. Such granule membranes are abundant in the young and adult "C" cell and would be reactive, while the small dense-core granules are prevalent in the fetus and would be unreactive, confirming the hypothesis of a difference of structure of the granule membrane and not of calcitonin between fetal and late postnatal "C" cells.