Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
권호 표지

Rho-associated Kinase is Involved in Preimplantation Development and Embryonic Compaction in Pigs

  • Received : 2010.05.15
  • Accepted : 2010.05.26
  • Published : 2010.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

The first morphogenetic event of preimplantation development, compaction, was required efficient production of porcine embryos in vitro. Compaction of the porcine embryo, which takes place at post 4-cell stage, is dependent upon the adhesion molecule E-cadherin. The E-cadherin through ${\beta}$-catenin contributes to stable cell-cell adhesion. Rho-associated kinase (ROCK) signaling was found to support the integrity of E-cadherin based cell contacts. In this study, we traced the effects of ROCK-1 on early embryonic development and structural integrity of blastocysts in pigs. Then, in order to gain new insights into the process of compaction, we also examined whether ROCK-1 signaling is involved in the regulation of the compaction mediated by E-cadherin of cellular adhesion molecules. As a result, real-time RT-PCR analysis showed that the expression of ROCK-1 mRNA was presented throughout porcine preimplantation stages, but not expressed as consistent levels. Thus, we investigated the blastocyst formation of porcine embryos treated with LPA and Y27632. Blastocysts formation and their qualities in LPA treated group increased significantly compared to those in the Y27632-treated group (p < 0.05). Then, to determine whether ROCK-1 associates embryonic compaction, we explored the effect of activator and/or inhibitor of ROCK-1 on compaction of embryos in pigs. The rate of compacted morula in LPA treated group was increased compared to that in the Y27632-treated group (39.7 vs 12.0%). Furthermore, we investigated the localization and expression pattern of E-cadherin at 4-cell stage porcine embryos in both LPA- and Y27632-treated groups by immunocytochemical analysis and Western blot analysis. The expression of E-cadherin was increased in LPA-treated group compared to that in the Y27632-treated group. The localization of E-cadherin in LPA-treated group was enriched in part of blastomere contacts compared to that Y27632-treated group. ROCK-1 as a crucial mediator of embryo compaction may plays an important role in regulating compaction through E-cadherin of the cell adhesion during the porcine preimplantation embryo. We concluded that ROCK-1 gene may affect the developmental potential of porcine blastocysts through regulating embryonic compaction.

Keywords

References

  1. Abeydeera LR and Day BN. 1997. Fertilization and subsequent development in vitro of pig oocytes inseminated in a modified tris-buffered medium with frozen-thawed ejaculated spermatozoa. BioI. Reprod. 57:729-734. https://doi.org/10.1095/biolreprod57.4.729
  2. Abeydeera LR. 2002. In vitro production of embryos in swine. Theriogenology 57:256-273.
  3. Butz S and Larue L. 1995. Expression of catenins during mouse embryonic development and in adult tissues. Cell Adhes. Commun. 3:337-352. https://doi.org/10.3109/15419069509081018
  4. Clayton L, Hall A and Johnson MH. 1999. A role for Rho-like GTPases in the polarisation of mouse eight-cell blastomeres. Dev. BioI. 205:322-331. https://doi.org/10.1006/dbio.1998.9117
  5. Hyafil F, Morello D, Babinet C and Jacob F. 1980. A cell surface glycoprotein involved in the compaction of embryonal carcinoma cells and cleavage stage embryos. Cell 21:927-934. https://doi.org/10.1016/0092-8674(80)90456-0
  6. Kawagishi R, Tahara M, Sawada K, Ikebuchi Y, Morishige K, Sakata M, Tasaka K and Murata Y. 2004. Rho-kinase is involved in mouse blastocyst cavity formation. Biochem. Biophys. Res. Commun. 319:643-648. https://doi.org/10.1016/j.bbrc.2004.05.040
  7. Kemler R, Babinet C, Eisen H and Jacob F. 1977. Surface antigen in early differentiation. Proc. Natl. Acad. Sci. USA 74:4449-4452. https://doi.org/10.1073/pnas.74.10.4449
  8. Kishi K, Sasaki T, Kuroda S, Itoh T and Takai Y. 1993. Regulation of cytoplasmic division of Xenopus embryo by rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI). J. Cell. BioI. 120:1187-1195. https://doi.org/10.1083/jcb.120.5.1187
  9. Leung T, Manser E, Tan L and Lim L. 1995. A novel serinel threonine kinase binding the Ras-related RhoA GTPase which translocates the kinase to peripheral membranes. J. BioI. Chem. 270:29051-29054. https://doi.org/10.1074/jbc.270.49.29051
  10. Machaty Z, Day BN and Prather RS. 1998. Development of early porcine embryos in vitro and in vivo. BioI. Reprod. 59:451-455. https://doi.org/10.1095/biolreprod59.2.451
  11. Noma K, Oyama N and Liao JK. 2006. Physiological role of ROCKs in the cardiovascular system. Am. J. Physiol. Cell Physiol. 290:C661-668. https://doi.org/10.1152/ajpcell.00459.2005
  12. Ohsugi M, Hwang SY, Butz S, Knowles BB, Solter D and Kemler R. 1996. Expression and cell membrane localization of catenins during mouse preimplantation development. Dev. Dyn. 206:391-402. https://doi.org/10.1002/(SICI)1097-0177(199608)206:4<391::AID-AJA5>3.0.CO;2-D
  13. Ohsugi M, Larue L, Schwarz H and Kemler R. 1997. Cell-junctional and cytoskeletal organization in mouse blastocysts lacking E-cadherin. Dev. BioI. 185:261-271. https://doi.org/10.1006/dbio.1997.8560
  14. Paterson HF, Self AJ, Garrett MD, Just I, Aktories K and Hall A. 1990. Microinjection of recombinant p21rho induces rapid changes in cell morphology. J. Cell. BioI. 111:1001-1007. https://doi.org/10.1083/jcb.111.3.1001
  15. Ridley AJ and Hall A. 1992. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell 70:389-399. https://doi.org/10.1016/0092-8674(92)90163-7
  16. Riento K and Ridley AJ. 2003. Rocks: multifunctional kinases in cell behaviour. Nat. Rev. Mol. Cell Biol. 4:446-456. https://doi.org/10.1038/nrm1128
  17. Takaishi K, Sasaki T, Kato M, Yamochi W, Kuroda S, Nakamura T, Takeichi M and Takai Y. 1994. Involvement of Rho p21 small GTP-binding protein and its regulator in the HGF-induced cell motility. Oncogene 9:273-279.
  18. Tao J, Tamis R, Fink K, Williams B, Nelson-White T and Craig R. 2002. The neglected morula/compact stage embryo transfer. Hum. Reprod. 17:1513-1518. https://doi.org/10.1093/humrep/17.6.1513
  19. Tominaga T, Sugie K, Hirata M, Morii N, Fukata J, Uchida A, Imura H and Narumiya S. 1993. Inhibition of PMA-induced, LFA-l-dependent lymphocyte aggregation by ADP ribosylation of the small molecular weight GTP binding protein, rho. J. Cell. BioI. 120:1529-1537. https://doi.org/10.1083/jcb.120.6.1529
  20. Tsukita S, Tsukita S, Nagafuchi A and Yonemura S. 1992. Molecular linkage between cadherins and actin filaments in cell-cell adherens junctions. Current Opinion in Cell Biology 4:834-839. https://doi.org/10.1016/0955-0674(92)90108-O
  21. Vaezi A, Bauer C, Vasioukhin V and Fuchs E. 2002. Actin cable dynamics and Rho/Rock orchestrate a polarized cytoskeletal architecture in the early steps of assembling a stratified epithelium. Dev. Cell 3:367-381. https://doi.org/10.1016/S1534-5807(02)00259-9
  22. Vestweber D and Kemler R. 1985. Identification of a putative cell adhesion domain of uvomorulin. EMBO J. 4:3393-3398.
  23. Yagi T and Takeichi M. 2000. Cadherin superfamily genes: functions, genomic organization, and neurologic diversity. Genes Dev. 14:1169-1180.