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Analysis of Osteopontin at the Maternal-Placental Interface in Pigs¹

^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

Noninvasive, epitheliochorial placentation in the pig follows a prolonged preimplantation period characterized by migration, spacing and elongation of conceptuses, and secretion of estrogen for maternal recognition of pregnancy. Osteopontin (OPN) is an extracellular matrix protein that binds integrins to promote cell-cell attachment and communication. OPN appears to play a key role in conceptus implantation and maintenance of pregnancy in sheep; however, a role for OPN in the porcine uterus has not been established. Therefore, this study examined OPN expression and function in the porcine uterus and conceptus (embryo/fetus and associated extraembryonic membranes). Northern and slot blot hybridization detected an increase in endometrial OPN expression between Days 25 and 30, and levels remained elevated through Day 85 of pregnancy. In situ hybridization localized OPN mRNA to discrete regions of the uterine luminal epithelium (LE) on Day 15 of pregnancy and to the entire LE thereafter. Glandular epithelial (GE) expression of OPN mRNA was first detected on Day 35 of pregnancy and increased through Day 85. Both 70- and 45-kDa forms of OPN protein were detected in cyclic and pregnant endometrium by Western blotting. OPN protein was localized to the LE and GE by immunofluorescence; however, only the 70-kDa OPN was detected in uterine flushings. OPN protein was present along the entire uterine-placental interface after Day 30 of pregnancy. In addition, OPN mRNA and protein were localized to immune-like cells within the stratum compactum of the endometrium in both Day 9 cyclic and pregnant gilts. Incubation of OPN-coated microbeads with porcine trophectoderm and uterine luminal epithelial cells induced Arg-Gly-Asp (RGD)-dependent integrin activation and transmembrane accumulation of cytoskeletal molecules at the apical cell surface as assessed by immunofluorescence detection of talin or -actinin as markers for focal adhesions. These results suggest that OPN, expressed by uterine epithelium and immune cells, may interact with receptors (i.e., integrins) on conceptus and uterus to promote conceptus development and signaling between these tissues as key contributors to attachment and placentation in the pig.

First decision: 23 August 2001.

¹ Research supported by USDA-NRICGP grant 95-35203-6337 to F.W.B. and R.C.B., USDA-NRICGP grant 95-35203-6223 to R.C.B., and NIH 1-F32-HD08501-01A1 to G.A.J.

² Correspondence: Fuller W. Bazer, Department of Animal Science and Center for Animal Biotechnology and Genomics, 442 Kleberg Center, Texas A&M University, College Station, TX 77843-2471. FAX: 979-862-2662; fbazer{at}cvm.tamu.edu

³ Current address: Department of Anatomy and Cell Biology, University of Kansas School of Medicine, Kansas City, KS 66160-7400

⁴ Current address: Department of Animal and Veterinary Science, Center for Reproductive Biology, University of Idaho, Moscow, ID 83844-2330

⁵ Equal contribution by both authors

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