Ovine osteopontin: II. Osteopontin and alpha(v)beta(3) integrin expression in the uterus and conceptus during the periimplantation period.
Osteopontin (OPN) is an acidic 70-kDa glycoprotein that is cleaved by proteases to yield 45-kDa and 24-kDa fragments. The 70-kDa and 45-kDa proteins contain a Gly-Arg-Gly-Asp-Ser (GRGDS) sequence that binds to cell surface integrins (primarily alpha(v)beta(3) heterodimer) to promote cell-cell attachment and cell spreading. A 70-kDa acidic protein was previously detected by two-dimensional (2D) PAGE in Day 17 pregnant endometrial cytosolic extracts using Stainsall and identified as immunoreactive OPN using Western blotting. Three forms of immunoreactive OPN proteins (70, 45, and 24 kDa) were detected by 1D PAGE and Western blot analysis of endometrial extracts. OPN protein in endometrial extracts did not differ between cyclic and pregnant ewes. However, the amount of 45-kDa OPN increased in uterine flushings from pregnant ewes between Days 11 and 17. Immunoreactive OPN was localized to luminal and glandular epithelia of both cyclic and pregnant ewes, and to trophectoderm of Day 19 conceptuses. The alpha(v) and beta(3) integrins were detected on Day 19 endometrium and conceptuses by immunofluorescence. It was reported that OPN mRNA increases in the uterine glands of pregnant ewes and secretion of OPN protein into the uterine lumen increases during early pregnancy. The present results demonstrate accumulation of OPN protein on endometrial LE and conceptus trophectoderm. Therefore, it is hypothesized that progesterone and/or interferon-tau induce expression, secretion and/or proteolytic cleavage of OPN by uterine epithelium. Secreted OPN is then available as ligand for alpha(v)beta(3) integrin heterodimer on trophectoderm and uterus to 1) stimulate changes in morphology of conceptus trophectoderm and 2) induce adhesion between luminal epithelium and trophectoderm essential for implantation and placentation.
Johnson
G A
GA
Department of Animal Science, Center for Animal Biotechnology and Genomics, Institute of Biosciences and Technology, Texas A&M University System Health Science Center, College Station, Texas 77843-2471, USA.
Burghardt
R C
RC
Spencer
T E
TE
Newton
G R
GR
Ott
T L
TL
Bazer
F W
FW
D000818:Animals; D015153:Blotting, Western / Q000662:veterinary; D017077:Culture Media, Conditioned; D046508:Culture Techniques; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004622:Embryo, Mammalian / Q000378:metabolism*; D047108:Embryonic Development; D004717:Endometrium / Q000378:metabolism; D005260:Female; D053495:Osteopontin; D011247:Pregnancy; D011270:Pregnancy, Animal / Q000378:metabolism*; D019030:Receptors, Vitronectin / Q000096:biosynthesis*; D012756:Sheep; D012795:Sialoglycoproteins / Q000096:biosynthesis*; D013997:Time Factors; D014599:Uterus / Q000378:metabolism*
D017077:Culture Media, Conditioned; D019030:Receptors, Vitronectin; D012795:Sialoglycoproteins; D053495:Osteopontin