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Biology of Reproduction 65, 820-828 (2001)
© 2001 Society for the Study of Reproduction, Inc.

Regular Article

Muc-1, Integrin, and Osteopontin Expression During the Implantation Cascade in Sheep1

Greg A. Johnson3,a,c, Fuller W. Bazera,c, Laurie A. Jaegera,b, Hakhyun Kaa,c, Jane E. Garlowa,c, Christiane Pfarrerd, Thomas E. Spencera,c, and Robert C. Burghardt2,a,b

a Center for Animal Biotechnology and Genomics, b Department of Veterinary Anatomy & Public Health, College of Veterinary Medicine c Department of Animal Science, Texas A&M University, College Station, Texas 77843-2471 d Department of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University, 35392 Giessen, Germany

ABSTRACT

The extracellular matrix protein osteopontin (OPN) is a component of histotroph that increases in uterine flushings from pregnant ewes during the peri-implantation period and is localized on the apical surfaces of the uterine luminal epithelium (LE) and conceptus trophectoderm (Tr). The potential involvement of OPN in the implantation adhesion cascade in sheep was investigated by examining temporal, spatial, and potential functional relationships between OPN, Muc-1, and integrin subunits during the estrous cycle and early pregnancy. Immunoreactive Muc-1 was highly expressed at the apical surfaces of uterine luminal (LE) and glandular epithelium (GE) in both cycling and pregnant ewes but was decreased dramatically on LE by Day 9 and was nearly undetectable by Day 17 of pregnancy when intimate contact between LE and Tr begins. In contrast, integrin subunits {alpha}v, {alpha}4, {alpha}5, ß1, ß3, and ß5 were constitutively expressed on conceptus Tr and at the apical surface of uterine LE and GE in both cyclic and early pregnant ewes. The apical expression of these subunits could contribute to the apical assembly of several OPN receptors including the {alpha}vß3, {alpha}vß1, {alpha}vß5, {alpha}4ß1, and {alpha}5ß1 heterodimers on endometrial LE and GE, and conceptus Tr in sheep. Functional analysis of potential OPN interactions with conceptus and endometrial integrins was performed on LE and Tr cells in vitro using beads coated with OPN, poly-L-lysine, or recombinant OPN in which the Arg-Gly-Asp sequence was replaced with RGE or RAD. Transmembrane accumulation of talin or {alpha}-actinin at the apical surface of uterine LE and conceptus Tr cells in contact with OPN-coated beads revealed functional integrin activation and cytoskeletal reorganization in response to OPN binding. These results provide a physiological framework for the role of OPN, a potential mediator of implantation in sheep, as a bridge between integrin heterodimers expressed by Tr and uterine LE responsible for adhesion for initial conceptus attachment.

FOOTNOTES

First decision: 2 April 2001.

1 Research supported by USDA-NRICGP grants 95-37203-2185 and 98-35203-6223 to F.W.B. and R.C.B. and by NIH 1-F32-HD08501-01A1 to G.A.J. Use of microscopy and imaging facilities in the College of Veterinary Medicine Image Analysis Laboratory, which is supported, in part, by NIH grant P30 ES09106, is acknowledged.

2 Correspondence: Robert C. Burghardt, Department of Veterinary Anatomy & Public Health, College of Veterinary Medicine, Texas A&M University, College Station, TX 77843. FAX: 979 847 8981; rburghardt{at}cvm.tamu.edu

3 Current address: Department of Animal and Veterinary Science, Center for Reproductive Biology, Agricultural Sciences Building, University of Idaho, Moscow, ID 83844-2330.




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