Morphological, histochemical and biochemical studies of the rat metrial gland

White, Linda Rosemary (1976) Morphological, histochemical and biochemical studies of the rat metrial gland. University of Southampton, Doctoral Thesis.

Abstract

The metrial gland a feature of the pregnant rodent uterus, was examined during mid-gestation in, rats both under normal conditions and after ovariecomy at the twelve day stage. Ovariectomy was found to induce marked impairment of development and extensive degeneration in the metrial gland, but despite the subsequent accumulation of cell debris and lipid, the invasion of macrophages normally observed after parturition was apparently absent. Ultrastructural and histochemical techniques were employed to study the sequence. of events.The most prominent feature of the metrial gland is the presence of large, granulated cells which may become binucleate and polyploid. Autoradiographic and microdensitometric studies indicated that at least 20% of metrial gland cell nuclei may become tetraploid, but few increase beyond octaploidy. It was not possible to determine how many of the measured nuclei belonged to ,granulated cells. Ovariectomy may reduce the differentiation of. some granulated cells, though many have already differentiated by the time the effects of ovariectomy become, prominent (about day 15). The cytoplasmic granules 'are larger than conventional lysosomes (about 1 pm)', but display several hydrolytic enzyme activities, including C-esterase(organ-phosphate and organ-mercurial resistant). After devising; a technique for the demonstration of C-esterase activity in cryostat, sections, a study of, these and other,esterases in the metrial gland was made,' though a comparison with C-esterase activity in.formalin-fixed frozen sections indicated that cryostat sections suffer from extensive elution of lyo-esterase which can be reduced by rapid formalin fixation. Initial characterisation of some of the esterase activities in cryostat sections was performed from observing the reaction to various activators and inhibitors. Further characterisation was then carried out by the use of polyacrilamide gradient gel electrophoresis to separate esterases from metrial gland homogenates. These results tended to confirm the initial histochemical findings. The major C-esterases associated with the metrial gland were clearly demonstrated; sub-fractionation studies suggested their location in the granules, and also indicated that they are not serum esterases although similar activity may be present in white blood cells. The two main.C-esterase bands demonstrated in the gels were calculated to have molecular weights of 64,000 and 58,000 daltons respectively, (the former is probably a glycoprotein). These estimations were sufficiently close to suggest that at least one of the.C-esterase' activities demonstrated may be , that of cathepsin D which is known. to exhibit similar activity and is localised in lysosomes. The isolation and biochemical studies which would be required to confirm this which, might include an assay technique developed during ,this study.

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