Mucus production by endometrium of reproductively healthy mares and mares with delayed uterine clearance

The aims of this study were to compare various staining and fixation techniques for endometrial biopsy samples and, thus, to investigate the mucociliary apparatus in endometrium from reproductively healthy mares and mares with delayed uterine clearance. Endometrial samples were collected from the left and right uterine horns of reproductively healthy mares (n=5) and mares with delayed uterine clearance (n=4) during anoestrus, transition, oestrus and dioestrus. Each sample of endometrium was fixed in either Bouin's fixative or formalin, and stained with alcian blue (pH 1.0 or 2.5), periodic acid Schiff (alone and in combination with alcian blue) or mucicarmine. An extracellular mucus blanket was observed more frequently in tissue fixed in Bouin's fixative than in formalin-fixed tissue. Luminal epithelial cells and the extracellular mucus blanket were stained preferentially using alcian blue (pH 2.5). Ciliated cells were observed most readily in biopsy samples fixed with formalin and stained with either alcian blue (pH 2.5) or mucicarmine. Mucus production was increased in fibrotic nests and inflamed endometrial samples. According to image analysis, the amount of intracellular mucus increases during oestrus, but secretions become denser during dioestrus (P < 0.05). In the present study, mucus production in mares with delayed uterine clearance was greater than that of reproductively healthy mares (P < 0.05).