Immunohistochemical and histochemical identification of proteins and carbohydrates in the equine endometrium: Expression patterns for mares suffering from endometrosis

Abstract

Although alterations in patterns of protein secretion revealed in uterine flushings from mares suffering from endometrosis have been described, little is known about alterations at the cellular level. Hence, the aim of this study was to characterize deviations in patterns of uterine gland secretion patterns using endometrial biopsies, histochemical and newly established immunohistochemical methods. Forty-eight endometrial biopsies were obtained from mares suffering from various types of endometrosis (active and inactive, destructive and non-destructive) and degree (mild to severe) were analyzed for expression of the proteins uteroglobin, uteroferrin, calbindin D9k and uterocalin as representatives of endometrial proteins detectable by immunohistochemistry using polyclonal antibodies. Glycogen was identified using the PAS-reaction and mucopolysaccharides were stained with alcian blue. Uterine glandular epithelia within fibrotic foci mostly revealed a protein and carbohydrate pattern of expression which was independent of hormonal changes during the estrous cycle. In comparison to non-affected glands, most epithelial cells within periglandular fibrosis exhibited decreased immunostaining intensity for proteins, especially when there was destructive endometrosis. However, uteroferrin staining intensity was strong within areas of severe destructive endometrosis. Moreover, only few basal glandular epithelial cells, especially those in cystic glands, stained for mucopolysaccharides that are typically seen within the luminal epithelia. Usually a single fibrotic focus caused only slight alterations in glandular proteins and carbohydrate reaction patterns, so that only more severe endometrosis lead to deviations which were detectable in uterine flushings. The highly sensitive methods used in the present study allow studies of uterine secretion patterns in the context of routine assessment of endometrial biopsies.