Öz
Kalsiyum bağımlı hücre adezyon moleküllerinden
kadherin üst ailesinin bir üyesi olan E-kadherin’in erken embriyonik gelişim
sırasında hücre adezyonunda kritik rolleri vardır. E-kadherin ekspresyonunu ve
dağılımını belirlemek için, geç 8 hücre, morula ve blastosist aşamalarında
bulunan embriyolara immünofloresan boyama uygulandı. İlk olarak dişi farelere
superovulasyon uyguladıktan sonra sakrifiye edilerek oviduktar izole edildi.
CD1 dişi farelerin oviduktları FHM HEPES-tamponlu medyum ile yıkanarak 2 hücreli
embriyo elde edildi ve istenilen embriyo aşamalarında kullanılmak üzere KSOM
medyumu içerisinde blastosist aşamasına kadar kültüre edildi. Embriyolar tespit
edildi ve E-kadherin ile immun boyama yapıldıktan sonra, floresan mikroskobu
kullanılarak görüntüler çekildi ve değerlendirildi. E-kadherinin geç 8 hücreli
aşamadan tamamen genişletilmiş blastosist aşamaları boyunca eksprese edildiği
gösterildi. Geç 8 hücreli embriyoda E-kadherin, membranın apikal bölgelerinde
yer almazken sadece hücre-hücre temas bölgelerinde var olduğu gözlemlendi.
Morulada E-kadherin, blastomerlerin yüzeyinde ve hücre-hücre birleşim
alanlarında lokalize olduğu görüldü. Blastosist aşamasındaki embriyolarda,
ICM'nin yüzeyinde lokalize olurken, TE hücrelerinde hücre-hücre temas bölgelerinde
membran boyaması olduğu da gösterilmiştir. Sonuçlarımız, E-cadherin
kaynaklı hücre adezyonunun, kompaksiyon, trofektoderm epitelyal farklılaşması
ve implantasyon sürecinde önemli rollere sahip olduğunu göstermektedir.
Anahtar Kelimeler
e-kadherin embriyo gelişimi fare hücre adezyonu preimplantasyon
Kaynakça
- De Vries, WN, Evsikov AV, Haac BE, Fancher KS, Holbrook AE, Kemler R, Solter D, Knowles BB (2004): Maternal beta-catenin and Ecadherin in mouse development. Development, 131, 4435-4445.
- Eckert JJ, Fleming TP (2008): Tight junction biogenesis during early development. Biochim Biophys Acta, 1778(3), 717-28.
- Fleming TP, Sheth B, Fesenko I (2001): Cell adhesion in the preimplantation mammalian embryo and its role in trophectoderm differentiation and blastocyst morphogenesis. Front Biosci, 6, D1000-7.
- Johnson MH, Maro B, Takeichi M (1986): The role of cell adhesion in the synchronisation and orientation of polarization in 8-cell mouse blastomeres. J Embryol Exp Morphol, 93, 239-255.
- Larue L, Ohsugi M, Hirchenhain J, Kemler R (1994): Ecadherin null mutant embryos fail to form a trophectoderm epithelium. Proc. Natl Acad. Sci. USA, 91, 8263–8267.
- Mihajlović AI, Bruce AW (2017): The first cell-fate decision of mouse preimplantation embryo development: integrating cell position and polarity. Open Biol, 7(11), pii: 170210. 70210.
- Mutluay D (2016): Distribution of primitive endoderm and epiblast lineage specific factors in late stage blastocysts Slov Vet Res, 53 (4), 211-7.
- Niessen CM, Gottardi CJ (2008): Molecular components of the adherens junction. Biochim Biophys Acta, 1778(3), 562-71.
- Ohsugi M, Hwang SY, Butz S, Knowles BB, Solter D, Kemler R (1996): Expression and cell membrane localization of catenins during mouse preimplantation development. Dev Dyn, 206(4), 391-402.
- Ohsugi M, Larue L, Schwarz H, Kemler R (1997): Cell-junctional and cytoskeletal organization in mouse blastocysts lacking E-cadherin. Dev Biol, 185(2), 261-71.
- Paria BC, Zhao X, Das SK, Dey SK, Yoshinaga K (1999): Zonula occludens-1 and E-cadherin are coordinately expressed in the mouse uterus with the initiation of implantation and decidualization. Dev Biol, 208(2), 488-501.
- Riethmacher D, Brinkmann V, Birchmeier C (1995): A targeted mutation in the mouse E-cadherin gene results in defective preimplantation development. Proc. Natl Acad. Sci. USA, 92, 855–859.
- Stephenson RO, Yamanaka Y, Rossant J (2010): Disorganized epithelial polarity and excess trophectoderm cell fate in preimplantation embryos lacking E-cadherin. Development, 137(20), 3383-91.
- Vestweber D, Gossler A, Boller K, Kemler R (1987): Expression and distribution of cell adhesion molecule uvomorulin in mouse preimplantation embryos. Dev Biol, 124(2), 451-6.
- Yamanaka Y, Ralston A, Stephenson RO, Rossant J (2006): Cell and molecular regulation of the mouse blastocyst. Dev Dyn. 235(9), 2301-14.
Öz
E-cadherin is a member of the cadherin superfamily of calcium-dependent cell adhesion molecules, and has critical roles in cell adhesion during early embryo development. To determine the expression and distribution of E-cadherin, we applied immunofluorescent staining on late 8 cell, morula and blastocyst stage embryos during the experiments. Firstly, we superovulated the females and sacrificed them to isolate oviducts. We flushed 2 cell embryos by using FHM HEPES-buffered medium from the CD1 female mouse oviducts and cultured them in KSOM medium until the blastocyst stage. Embryos were fixed and immunostained with E-cadherin, then images were assessed using a fluorescence microscope. We demonstrate that E-cadherin is expressed throughout the late 8 cell to fully expanded blastocysts. We observed that E-cadherin is present only in the cell-cell contact sites and not in the apical sites of the membrane of late 8 cell stage embryo. In morula, E-cadherin localized on the surface of blastomeres and cell-cell junctions. In blastocyst stage, embryos showed membrane staining in cell-cell contact regions in trophectoderm cells while localized on the surface of inner cell mass. Our results indicate that E-cadherin mediated cell adhesion has major roles on compaction, trophectoderm epithelial differentation and implantation.
Anahtar Kelimeler
Cell adhesion e-cadherin embryo development mouse preimplantation
Kaynakça
- De Vries, WN, Evsikov AV, Haac BE, Fancher KS, Holbrook AE, Kemler R, Solter D, Knowles BB (2004): Maternal beta-catenin and Ecadherin in mouse development. Development, 131, 4435-4445.
- Eckert JJ, Fleming TP (2008): Tight junction biogenesis during early development. Biochim Biophys Acta, 1778(3), 717-28.
- Fleming TP, Sheth B, Fesenko I (2001): Cell adhesion in the preimplantation mammalian embryo and its role in trophectoderm differentiation and blastocyst morphogenesis. Front Biosci, 6, D1000-7.
- Johnson MH, Maro B, Takeichi M (1986): The role of cell adhesion in the synchronisation and orientation of polarization in 8-cell mouse blastomeres. J Embryol Exp Morphol, 93, 239-255.
- Larue L, Ohsugi M, Hirchenhain J, Kemler R (1994): Ecadherin null mutant embryos fail to form a trophectoderm epithelium. Proc. Natl Acad. Sci. USA, 91, 8263–8267.
- Mihajlović AI, Bruce AW (2017): The first cell-fate decision of mouse preimplantation embryo development: integrating cell position and polarity. Open Biol, 7(11), pii: 170210. 70210.
- Mutluay D (2016): Distribution of primitive endoderm and epiblast lineage specific factors in late stage blastocysts Slov Vet Res, 53 (4), 211-7.
- Niessen CM, Gottardi CJ (2008): Molecular components of the adherens junction. Biochim Biophys Acta, 1778(3), 562-71.
- Ohsugi M, Hwang SY, Butz S, Knowles BB, Solter D, Kemler R (1996): Expression and cell membrane localization of catenins during mouse preimplantation development. Dev Dyn, 206(4), 391-402.
- Ohsugi M, Larue L, Schwarz H, Kemler R (1997): Cell-junctional and cytoskeletal organization in mouse blastocysts lacking E-cadherin. Dev Biol, 185(2), 261-71.
- Paria BC, Zhao X, Das SK, Dey SK, Yoshinaga K (1999): Zonula occludens-1 and E-cadherin are coordinately expressed in the mouse uterus with the initiation of implantation and decidualization. Dev Biol, 208(2), 488-501.
- Riethmacher D, Brinkmann V, Birchmeier C (1995): A targeted mutation in the mouse E-cadherin gene results in defective preimplantation development. Proc. Natl Acad. Sci. USA, 92, 855–859.
- Stephenson RO, Yamanaka Y, Rossant J (2010): Disorganized epithelial polarity and excess trophectoderm cell fate in preimplantation embryos lacking E-cadherin. Development, 137(20), 3383-91.
- Vestweber D, Gossler A, Boller K, Kemler R (1987): Expression and distribution of cell adhesion molecule uvomorulin in mouse preimplantation embryos. Dev Biol, 124(2), 451-6.
- Yamanaka Y, Ralston A, Stephenson RO, Rossant J (2006): Cell and molecular regulation of the mouse blastocyst. Dev Dyn. 235(9), 2301-14.
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Veteriner Cerrahi |
| Bölüm | Araştırma Makalesi |
| Yazarlar | |
| Yayımlanma Tarihi | 15 Ocak 2020 |
| Gönderilme Tarihi | 18 Haziran 2019 |
| Kabul Tarihi | 3 Eylül 2019 |
| Yayımlandığı Sayı | Yıl 2020 Cilt: 91 Sayı: 1 |
Veteriner Hekimler Derneği Dergisi açık erişimli bir dergi olup, derginin yayın modeli Budapeşte Erişim Girişimi (BOAI) bildirisine dayanmaktadır. Yayınlanan tüm içerik, çevrimiçi ve ücretsiz olarak sunulan Creative Commons CC BY-NC 4.0 lisansı altında lisanslanmıştır. Yazarlar, Veteriner Hekimler Derneği Dergisi'nde yayınlanan eserlerinin telif haklarını saklı tutarlar.
Veteriner Hekimler Derneği / Turkish Veterinary Medical Society