Expression of Carbohydrate Antigens in the Goat Uterus During Early Pregnancy and on Steroid-Treated Polarized Uterine Epithelial Cells In Vitro1

Abstract

Our objectives were to determine whether specific fucosylated carbohydrate antigens, associated with uterine receptivity in rodents, are expressed in pregnant caprine uterine tissues and polarized uterine luminal epithelial (ULE) cells in culture. Immunofluorescence microscopy on frozen endometrium revealed that expression of the H-type 1 antigen, confined to epithelial cells, was regulated during early pregnancy. Staining was high on Day 5 and low on Days 11 and 13. Strong, uniform apical staining was characteristic of ULE cells between Days 15 and 19 but declined markedly by Day 25. Immunofluorescence analysis of the apical surface of polarized ULE cells cultured in steroid-free medium revealed weak and diffuse staining for the H-type 1 antigen, while progesterone (P(4)) treatment resulted in the formation of aggregates of punctate staining along the apical surface. Domain-specific biotinylation of polarized ULE cells, coupled with streptavidin precipitation and Western blotting, revealed that six apical surface proteins (31, 33, 42, 55, 60, and 70 kDa) carry the H-type 1 antigen. Therefore, H-type 1 antigen expression is up-regulated in vivo during the periimplantation period, stimulated by P(4) on polarized ULE cells in culture, and may be a useful marker for uterine receptivity in this species.

Implantation and Establishment of Pregnancy in the Pig

“…However, it is likely that fi lamentous porcine conceptuses undergo a series of attach-andrelease events between carbohydrates and lectin receptors at the apical surfaces of trophectoderm and uterine LE that result in maximal apposition of these tissues, similar to the "rolling and tethering" that occur during leukocyte adhesion to the endothelium for extravasation of leukocytes (Kling et al 1992 ), and proposed for the initial attachment of human blastocysts to the uterine wall (Red-Horse et al 2004 ). In support of this idea, other domestic farm species, goats and sheep, exhibit prominent expression of H-type 1 antigens and glycosylation-dependent glycam 1, respectively, at the uterine-conceptus interface during implantation (Powell et al 2000 ;Gray et al 2004 ). These low-affi nity contacts are then replaced by a more stable and extensive repertoire of adhesive interactions between integrins and maternal ECM which appear to be the major contributors to stable adhesion at implantation (Hynes 1987 ;Ruoslahti and Pierschbacher 1987 ;Aplin et al 1994 ;Burghardt et al 1997Burghardt et al , 2002Johnson et al 2001 ;Lessey 2002 ).…”

Matrix-Metalloproteinases-2 and -9 Production in Bovine Endometrial Cell Culture.

“…It was noted that MMP-2 production was similar both in non-collagen coated and collagen coated dishes. It is known that cell polarity affects protein secretion,but it was reported that cells could be correctly polarized after high-density plating [22,23]. It is possible that MMP-2 is produced by endometrial epithelial cells [17], although some previous reports indicated that endometrial epithelial cells do not produce MMP-2 [10,24].…”

Uterine receptivity and the plasma membrane transformation

“…Much work on carbohydrates has identified several epitopes which are up-regulated in different species around the time of blastocyst attachment and which have in common numbers of glucosamine and fucose residues [74][75][76][77][78][79][80]. There is in addition emerging evidence that some integrins, cadherins and related receptors may be up-regulated in different membrane compartments in species so far examined which includes rodents, pigs, rabbits and humans during early pregnancy [14,63,[81][82][83][84][85].…”