Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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Analysis of the Gene Expression by Laser Capture Microdissection (III) -Microarray Analysis of the Gene Expression at the Mouse Uterine Luminal Epithelium of the Implantation Sites during Apposition Period-

Laser Capture Microdissection을 이용한 유전자 발현 연구 (III) -생쥐 착상 부위 자궁 내강상피 조직에서 배아 병치 기간 동안 일어나는 유전자 발현에 관한 Microarray 분석-

  • Yoon, Se-Jin (Infertility Medical Center, CHA General Hospital) ;
  • Jeon, Eun-Hyun (Graduate School of Life Science and Biotechnology, College of Medicine, Pochon CHA University) ;
  • Park, Chang-Eun (Infertility Medical Center, CHA General Hospital) ;
  • Ko, Jung-Jae (Infertility Medical Center, CHA General Hospital) ;
  • Choi, Dong-Hee (Infertility Medical Center, CHA General Hospital) ;
  • Cha, Kwang-Yul (Infertility Medical Center, CHA General Hospital) ;
  • Kim, Se-Nyun (Digital Genomics Inc.) ;
  • Lee, Kyung-Ah (Infertility Medical Center, CHA General Hospital)
  • 윤세진 (차병원 여성의학연구소) ;
  • 전은현 (포천중문 의과대학교 생명과학 전문대학원) ;
  • 박창은 (차병원 여성의학연구소) ;
  • 고정재 (차병원 여성의학연구소) ;
  • 최동희 (차병원 여성의학연구소) ;
  • 차광열 (차병원 여성의학연구소) ;
  • 김세년 (디지탈 지로믹스) ;
  • 이경아 (차병원 여성의학연구소)
  • Published : 2002.12.30
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Abstract

Object: The present study was accomplished to obtain a gene expression profile of the luminal epithelium during embryo apposition in comparison of implantation (1M) and interimplantation (INTER) sites. Material and Method: The mouse uterine luminal epithelium from IM and INTER sites were sampled on day 4.5 (Day of vaginal plug = day 0.5) by Laser Captured Microdissection (LCM). RNA was extracted from LCM captured epithelium, amplified, labeled and hybridized to microarrays. Results from microarray hybridization were analyzed by Significance Analysis of Microarrays (SAM) method. Differential expression of some genes was confirmed by LCM followed by RT-PCR. Results: Comparison of IM and INTER sites by SAM identified 73 genes most highly ranked at IM, while 13 genes at the INTER sites, within the estimated false discovery rate (FDR) of 0.163. Among 73 genes at IM, 20 were EST/unknown function, and the remain 53 were categorized to the structural, cell cycle, gene/protein expression, immune reaction, invasion, metabolism, oxidative stress, and signal transduction. Of the 24 structural genes, 14 were related especially to extracellular matrix and tissue remodeling. Meanwhile, among 13 genes up-regulated at INTER, 8 genes were EST/unknown function, and the rest 5 were related to metabolism, signal transduction, and gene/protein expression. Among these 58 (53+5) genes with known functions, 13 genes (22.4%) were related with $Ca^{2+}$ for their function. Conclusions: Results of the present study suggest that 1) active tissue remodeling is occurring at the IM sites during embryo apposition, 2) the INTER sites are relatively quiescent than IM sites, and 3) the $Ca^{2+}$ may be a crucial for apposition. Search for human homologue of those genes expressed in the mouse luminal epithelium during apposition will help to understand the implantation process and/or implantation failure in humans.

Keywords

References

  1. Paria BC, Huet-Hudson YM, Dey SK. Blastocyst's state of activity determines the 'window' of implantation in the receptive mouse uterus. Proc Natl Acad Sci 1993;90: 10159-62 https://doi.org/10.1073/pnas.90.21.10159
  2. Paria BC, Lim H, Das SK, Reese J, Dey SK. Molecular signaling in uterine receptivity for implantation. Cell Dev BioI 2000; 11: 67-76 https://doi.org/10.1006/scdb.2000.0153
  3. Shiotani M, Noda Y, Mori T. Embryo-dependent induction of uterine receptivity assessed by an in vitro model of implantation in mice. BioI Reprod 1993; 49: 794-801 https://doi.org/10.1095/biolreprod49.4.794
  4. Simon C, Martin JC, Galan A, Valbuena D, Pellicer A. Embryonic regulation in implantation. Seminars Reprod Endocrinol 1999; 17:267-74 https://doi.org/10.1055/s-2007-1016234
  5. Reese J, Das SK, Paria BC, Lim H, Song H, Matsumoto H, et al. Global gene expression analysis to identify molecular markers of uterine receptivity and embryo implantation. J BioI Chem 2001; 276: 44137-45 https://doi.org/10.1074/jbc.M107563200
  6. Carson DD, Lagow E, Thathiah A, Al-Shami R, Farach-Carson MC, Vernon M, et al. Changes in gene expression during the early to mid-luteal (receptive phase) transition in human endometrium detected by high-density microarray screening. Mol Human Reprod 2002; 8: 871-9 https://doi.org/10.1093/molehr/8.9.871
  7. Kao LC, Tulac S, Lobo S, Imani B, Yang JP, Germeyer A, et al. Global gene profiling in human endometrium during the window of implantation. Endocrinol 2002; 143: 2119-38 https://doi.org/10.1210/en.143.6.2119
  8. Yoshioka K, Matsuda F, Takakura K, Noda Y, Imakawa K, Sakai S. Determination of genes involved in the process of implantation: application of Gene Chip to scan 6500 genes. Biochem Biophys Res Commun 2000;272: 531-8 https://doi.org/10.1006/bbrc.2000.2818
  9. Nie GY, Hampton AL, Salamonsen LA, Clements JA, Findlay JK. Identification of monoclonal nonspecific suppressor factor beta (mNSFbeta) as one of the genes differentially expressed at implantation sites compared to interimplantation sites in the mouse uterus. Mol Reprod Dev 2000; 55: 351-63 https://doi.org/10.1002/(SICI)1098-2795(200004)55:4<351::AID-MRD1>3.0.CO;2-L
  10. Popovici RM, Kao L-C, Giudice LC. Discovery of new inducible genes in in vitro decidualized human endometrial stromal cells using microarray technology. Endocrinol 2000; 141: 3510-3 https://doi.org/10.1210/en.141.9.3510
  11. Salamonsen LA, Nie G, Findlay JK. Newly identified endometrial genes of importance for implantation. J Reprod Immunol 2002; 53: 215-25 https://doi.org/10.1016/S0165-0378(01)00087-0
  12. Bonner RF, Emmert BM, Cole K, Pohida T, Chuaqui R, Goldstein S, et al. Laser capture microdissection: Molecular analysis of tissue. Science 1997; 278: 1481-3 https://doi.org/10.1126/science.278.5342.1481
  13. Okulicz WC, Ace CI. Progesterone-regulated gene expression in the primate endometrium. Semin Reprod Endocrinol 1999; 17: 241-55 https://doi.org/10.1055/s-2007-1016232
  14. Yang YH, Dudoit S, Luu P, Lin DM, Peng V, Ngai J, et al. Normalization for cDNA microarray data: a robust composite method addressing single and multiple slide systematic variation. Nucl Acid Res 2002; 30: e15
  15. Tusher VG, Tibshirani R, Chu G. Significance analysis of microarrays applied to the ionizing radiation response. PNAS 2001; 98: 5116-21 https://doi.org/10.1073/pnas.091062498
  16. Murphy CR. The cytoskeleton of uterine epithelial cells: a new player in uterine receptivity and the plasma membrane transformation. Hum Reprod Update 1995; 1: 567-80 https://doi.org/10.1093/humupd/1.6.567
  17. Van der Rest M, Garrone R. Collagen family of proteins. FASEB J 1991; 5: 3814-23
  18. Dafforn TR, Della M, Miller AD. The molecular interactions of heat shock protein 47 (Hsp47) and their implications for collagen biosynthesis. J BioI Chem 2001; 276: 49310-9 https://doi.org/10.1074/jbc.M108896200
  19. Hendershot LM, Bulleid NJ. Protein-specific chaperones: the role of hsp47 begins to gel. Curr BioI 2000; 10: R912-915 https://doi.org/10.1016/S0960-9822(00)00850-2
  20. Sauk JJ, Smith T, Norris K, Ferreira L. Hsp47 and the translation-translocation machinery cooperate in the production of alpha 1 (I) chains of type 1 procollagen. J Biol Chem 1994; 269: 3941-6
  21. Satoh M, Hirayoshi K, Yokota SI, Hosokawa N, Nagata K. Intracellular interaction of collagen-specific stress protein HSP47 with newly synthesized procollagen. J Cell BioI 1996; 133: 469-83 https://doi.org/10.1083/jcb.133.2.469
  22. Yan Q, Sage EH. SPARC, a matricellular glycoprotein with important biological functions. J Histochem Cytochem 1999; 47: 1495-506 https://doi.org/10.1177/002215549904701201
  23. Brekken RA, Sage EH. SPARC, a matricellular protein: at the crossroads of cell-matrix communication. Matrix BioI 2001; 19: 816-27
  24. Sasakie T, Gohring W, Mann K, Maurer P, Hohenester E, Knauper V, et al. limited cleavage of extracellular matrix protein BM-40 by matrix metalloproteinases increases its affinity for collagens. J BioI Chem 1997; 272: 9237-43 https://doi.org/10.1074/jbc.272.14.9237
  25. Tsuchida K, Arai KY, Kuramoto Y, Yamakawa N, Hasegawa Y, Sugino H. Identification and characterization of a novel follistatin-like protein as a binding protein for the TGF-beta family. J BioI Chem 2000;275:40788-96 https://doi.org/10.1074/jbc.M006114200
  26. Evanas J, Malmendal A, Forsen S. Calcium. Curr Opin Chem BioI 1998; 2: 293-302 https://doi.org/10.1016/S1367-5931(98)80072-0
  27. Das SK, Tan J, Raja S, Halder J, Paria BC, Dey SK. Estrogen targets genes involved in protein processing, calcium homeostasis, and Wnt signaling in the mouse uterus independent of estrogen receptoralpha and -beta. J BioI Chem 2000; 275: 28834-42 https://doi.org/10.1074/jbc.M003827200
  28. Tinel H, Denker HW, Thie M. Calcium influx in human uterine epithelial RL95-2 cells triggers adhesiveness for trophoblast-like cells. Model studies on signaling events during embryo implantation. Mol Human Reprod 2000; 6: 1119-30 https://doi.org/10.1093/molehr/6.12.1119
  29. Hohn HP, Linke M, Denker HW Adhesion of trophoblast to uterine epithelium as related to the state of trophoblast differentiation: in vitro studies using cell lines. Mol Reprod Dev 2000; 57: 135-45
  30. Aplin JD. Adhesion molecules in implantation. Rev Reprod 1997; 2: 84-93 https://doi.org/10.1530/ror.0.0020084
  31. Aplin JD, Meseguer M, Simon C, Ortiz ME, Croxatto H, Jones CJP. MUC1, glycan and the cellsurface barrier to embryo implantation. Biochem Soc Trans 2001; 29: 153-6 https://doi.org/10.1042/BST0290153
  32. Sharkey A. Cytokines and implantation. Rev Reprod 1998; 3: 52-61 https://doi.org/10.1530/ror.0.0030052
  33. Stewart CL, Kasper P, Brunet LJ, Bhatt H, Gadi J, Kontgen F, et al. Blastocyst implantation depends on maternal expression of leukaemia inhibitory factor. Nature 1992; 359: 76-9 https://doi.org/10.1038/359076a0