Identification of Differentially Expressed Genes in the Uterine Endometrium on Day 12 of the Estrous Cycle and Pregnancy in Pigs

SUMMARY Maternal recognition of pregnancy in pigs occurs approximately on Day (D) 12 of pregnancy and is critical for embryo implantation. The presence of the conceptus in the uterine lumen during this period changes uterine endometrial function to prepare for attachment of the conceptus to the endometrial epithelial cells and maintain luteal functionintheovary.Althoughmuchisknownabout endometrialgeneexpression,the genes expressed in the uterine endometria and the cellular and molecular mechanisms of those gene products during the period of implantation and maternal recognition of pregnancy in pigs are still not completely defined. To better understand the interactions between the maternal uterus and conceptus during the implantation process, we searched genes differentially expressed in the endometria on D12 of pregnancy compared to those on D12 of the estrous cycle. A new reverse transcription-polymerase chain reaction (RT-PCR)-based method that involves annealing control primers (ACPs) was employed. Using 120 ACPs, we sequenced 12 differentially expressed genes (DEGs) and identified those genes using the Basic Local Alignment Search Tool (BLAST). Northern blot hybridization analysis confirmed the differential expression of those DEGs in the uterine endometrium. In situ hybridization analysis determined the cell-type specific expression of the DEGs in the uterine endometrium. Further analysis of the DEGs found in this study will provide insights into the cellular and molecular basis of maternal and fetal interactions during the period of maternal recognition of pregnancy in the pig. Mol. Reprod. Dev. 76:

[1]  T. Spencer,et al.  Functional analysis of autocrine and paracrine signalling at the uterine-conceptus interface in pigs. , 2020, Reproduction (Cambridge, England) Supplement.

[2]  J. Yelich,et al.  Regulation of conceptus development and attachment in pigs. , 2020, Journal of reproduction and fertility. Supplement.

[3]  F. Bazer,et al.  Extracellular matrix and the implantation cascade in pigs. , 2020, Journal of reproduction and fertility. Supplement.

[4]  R. Prather,et al.  The use of microarrays to define functionally-related genes that are differentially expressed in the cycling pig uterus. , 2019, Society of Reproduction and Fertility supplement.

[5]  T. Ruzicka,et al.  S100A15, an antimicrobial protein of the skin: regulation by E. coli through Toll-like receptor 4. , 2007, The Journal of investigative dermatology.

[6]  T. Ruzicka,et al.  Human S100A15 splice variants are differentially expressed in inflammatory skin diseases and regulated through Th1 cytokines and calcium , 2007, Experimental dermatology.

[7]  E. Jeung,et al.  Uterine TRPV6 expression during the estrous cycle and pregnancy in a mouse model. , 2007, American journal of physiology. Endocrinology and metabolism.

[8]  J. Hartwig,et al.  Modifications of cellular responses to lysophosphatidic acid and platelet-activating factor by plasma gelsolin. , 2007, American journal of physiology. Cell physiology.

[9]  Kyung-Chul Choi,et al.  Differential expression of uterine calcium transporter 1 and plasma membrane Ca2+ ATPase 1b during rat estrous cycle. , 2006, American journal of physiology. Endocrinology and metabolism.

[10]  Ji-Long Liu,et al.  Serial Analysis of Gene Expression in Mouse Uterus at the Implantation Site* , 2006, Journal of Biological Chemistry.

[11]  A. Iavarone,et al.  Id family of helix-loop-helix proteins in cancer , 2005, Nature Reviews Cancer.

[12]  C. V. Van Tassell,et al.  Serial analysis of gene expression during elongation of the peri-implantation porcine trophectoderm (conceptus). , 2005, Physiological genomics.

[13]  Kathleen C. Lee,et al.  S100 proteins in the epidermis. , 2004, The Journal of investigative dermatology.

[14]  U. Schumacher,et al.  Different transcriptional expression of KIAA1324 and its splicing variants in human carcinoma cell lines with different metastatic capacity. , 2004, Oncology reports.

[15]  I. Hwang,et al.  Annealing control primer system for identification of differentially expressed genes on agarose gels. , 2004, BioTechniques.

[16]  I. Hwang,et al.  Annealing control primer system for improving specificity of PCR amplification. , 2003, BioTechniques.

[17]  R. Blouin,et al.  Isolation of Differentially Expressed Genes in Conceptuses and Endometrial Tissue of Sows in Early Gestation1 , 2003, Biology of reproduction.

[18]  A. Mirmohammadsadegh,et al.  Molecular cloning and characterization of alternatively spliced mRNA isoforms from psoriatic skin encoding a novel member of the S100 family , 2003, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[19]  T. Spencer,et al.  Keratinocyte Growth Factor: Expression by Endometrial Epithelia of the Porcine Uterus , 2000, Biology of reproduction.

[20]  M. Israel,et al.  Id-2 regulates critical aspects of human cytotrophoblast differentiation, invasion and migration. , 2000, Development.

[21]  Helen L. Yin,et al.  Gelsolin, a Multifunctional Actin Regulatory Protein* , 1999, The Journal of Biological Chemistry.

[22]  A. Scaloni,et al.  Lipocalins of boar salivary glands binding odours and pheromones. , 1998, European journal of biochemistry.

[23]  F. Bazer,et al.  Spatial and temporal analyses of integrin and Muc-1 expression in porcine uterine epithelium and trophectoderm in vivo. , 1996, Biology of reproduction.

[24]  D R Flower,et al.  The lipocalin protein family: structure and function. , 1996, The Biochemical journal.

[25]  M. Desouza,et al.  Messenger RNA encoding an estrogen‐dependent oviduct secretory protein in the sheep is localized in the apical tips and basal compartments of fimbria and ampulla epithelial cells implying translation at unique cytoplasmic foci , 1995, Molecular reproduction and development.

[26]  Walter Witke,et al.  Hemostatic, inflammatory, and fibroblast responses are blunted in mice lacking gelsolin , 1995, Cell.

[27]  G. J. King,et al.  Effects of topical and systemic estrogen on morphology of porcine uterine luminal epithelia. , 1992, Biology of reproduction.

[28]  R. Blair,et al.  Effect of asynchronous transfer and oestrogen administration on survival and development of porcine embryos. , 1991, Journal of reproduction and fertility.

[29]  F. Bazer,et al.  Production of two species of interferon by Large White and Meishan pig conceptuses during the peri-attachment period. , 1991, Journal of Reproduction and Fertility.

[30]  M. Guillomot,et al.  Interferon‐gamma gene and protein are spontaneously expressed by the porcine trophectoderm early in gestation , 1990, European journal of immunology.

[31]  R. Roberts,et al.  Porcine conceptuses secrete an interferon during the preattachment period of early pregnancy. , 1989, Biology of reproduction.

[32]  G. Morgan,et al.  Development and survival of pig blastocysts after oestrogen administration on day 9 or days 9 and 10 of pregnancy. , 1987, Journal of reproduction and fertility.

[33]  S. Orkin,et al.  Plasma and cytoplasmic gelsolins are encoded by a single gene and contain a duplicated actin-binding domain , 1986, Nature.

[34]  J. Lawrence,et al.  Intracellular localization of messenger RNAs for cytoskeletal proteins , 1986, Cell.

[35]  R. Roberts,et al.  Establishment of pregnancy in the pig: III. Endometrial secretory response to estradiol valerate administered on day 11 of the estrous cycle. , 1982, Biology of reproduction.

[36]  R. Roberts,et al.  Establishment of pregnancy in the pig: I. Interrelationships between preimplantation development of the pig blastocyst and uterine endometrial secretions. , 1982, Biology of reproduction.

[37]  F. Bazer,et al.  Theory of maternal recognition of pregnancy in swine based on estrogen controlled endocrine versus exocrine secretion of prostaglandin F2alpha by the uterine endometrium. , 1977, Prostaglandins.

[38]  A. Enders,et al.  Cellular basis of interaction between trophoblast and uterus at implantation. , 1975, Biology of reproduction.

[39]  J. S. Perry,et al.  Steroid Hormone Production by Pig Blastocysts , 1973, Nature.

[40]  B. Nilius,et al.  Calcium absorption across epithelia. , 2005, Physiological reviews.

[41]  Amir Jazaeri,et al.  Microarray analysis reveals distinct gene expression profiles among different histologic types of endometrial cancer. , 2003, Cancer research.

[42]  L. Salamonsen,et al.  Newly identified endometrial genes of importance for implantation. , 2002, Journal of reproductive immunology.

[43]  C. Wood,et al.  Discovery and characterization of endometrial epithelial messenger ribonucleic acids using the ovine uterine gland knockout model. , 1999, Endocrinology.

[44]  T. Spencer,et al.  Endocrinology of the Transition from Recurring Estrous Cycles to Establishment of Pregnancy in Subprimate Mammals , 1998 .

[45]  W. Wahli,et al.  A Simplified In Situ Hybridization Protocol Using Non-radioactively Labeled Probes to Detect Abundant and Rare mRNAs on Tissue Sections , 1998 .