|
|
|
|
|
| Expression patterns of OCT4, NANOG, and SOX2 in goat preimplantation embryos from in vivo and in vitro |
| YU Xiao-li, ZHAO Xiao-e, WANG Hua-yan, MA Bao-hua |
1、College of Veterinary Medicine, Northwest A&F University, Yangling 712100, P.R.China
2、Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Yangling 712100, P.R.China |
|
|
|
摘要 The transcription factors, including OCT4, NANOG, and SOX2, played crucial roles in the maintenance of self-renewal and pluripotency in embryonic stem cells (ESCs). They expressed in preimplantation mammalian development with spatio- temporal pattern and took part in regulation of development. However, their expression and roles in goat had not been reported. In the present study, the expression of OCT4, NANOG, and SOX2 in goat preimplantation embryos both in vivo and in vitro were detected by real-time RCR and immunofluorescence. For in vivo fertilized embryos, the transcripts of OCT4, NANOG, and SOX2 could be detected from oocytes to blastocyst stage, their expression in morula and blastocyst stages was much higher than other stage. OCT4 protein was detected from oocyte to blastocyst, but the fluorescence was more located-intensive with nuclei from 8-cell stage, its expression present in both inner cell mass (ICM) and trophoblast cells (TE) at blastocyse stage. NANOG protein was similar to OCT4, the signaling of fluorescence completely focused on cell nuclei, while the SOX2 firstly showed nuclei location in morula. Comparing to in vivo fertilized embryo, the mRNA of these three transcription factors could be detected at 8-cell stage in parthenogenetic embryos (in vitro). Thereafter, the expressional level rose gradually along with embryo development. The locations of OCT4 and NANOG proteins were similar to in vivo fertilized embryos, and they located in cell nuclei from morula to blastocyst stage, while SOX2 protein firstly could be detected in cell nuclei at 8-cell stage. These differences suggested that OCT4, NANOG, and SOX2 played different function in regulating development of goat preimplantation embryos. These results may provide a novel insight to goat embryo development and be useful for goat ESCs isolation.
Abstract The transcription factors, including OCT4, NANOG, and SOX2, played crucial roles in the maintenance of self-renewal and pluripotency in embryonic stem cells (ESCs). They expressed in preimplantation mammalian development with spatio- temporal pattern and took part in regulation of development. However, their expression and roles in goat had not been reported. In the present study, the expression of OCT4, NANOG, and SOX2 in goat preimplantation embryos both in vivo and in vitro were detected by real-time RCR and immunofluorescence. For in vivo fertilized embryos, the transcripts of OCT4, NANOG, and SOX2 could be detected from oocytes to blastocyst stage, their expression in morula and blastocyst stages was much higher than other stage. OCT4 protein was detected from oocyte to blastocyst, but the fluorescence was more located-intensive with nuclei from 8-cell stage, its expression present in both inner cell mass (ICM) and trophoblast cells (TE) at blastocyse stage. NANOG protein was similar to OCT4, the signaling of fluorescence completely focused on cell nuclei, while the SOX2 firstly showed nuclei location in morula. Comparing to in vivo fertilized embryo, the mRNA of these three transcription factors could be detected at 8-cell stage in parthenogenetic embryos (in vitro). Thereafter, the expressional level rose gradually along with embryo development. The locations of OCT4 and NANOG proteins were similar to in vivo fertilized embryos, and they located in cell nuclei from morula to blastocyst stage, while SOX2 protein firstly could be detected in cell nuclei at 8-cell stage. These differences suggested that OCT4, NANOG, and SOX2 played different function in regulating development of goat preimplantation embryos. These results may provide a novel insight to goat embryo development and be useful for goat ESCs isolation.
|
|
Received: 30 July 2014
Accepted:
|
| Fund: This work was supported by the Genetically Modified Organisms Breeding Major Projects, Ministry of Agriculture, China (2008ZX0810-001). |
|
Corresponding Authors:
MABao-hua, Tel: +86-29-87091117, Fax: +86-29-87091032,E-mail: mabh@nwsuaf.edu.cn
E-mail: mabh@nwsuaf.edu.cn
|
| About author: YU Xiao-li, E-mail: yuxiaoli1221@gmail.com; |
Cite this article:
YU Xiao-li, ZHAO Xiao-e, WANG Hua-yan, MA Bao-hua.
2015.
Expression patterns of OCT4, NANOG, and SOX2 in goat preimplantation embryos from in vivo and in vitro. Journal of Integrative Agriculture, 14(7): 1398-1406.
|
| Avilion A A, Nicolis S K, Pevny L H, Perez L, Vivian N,Lovell-Badge R. 2003. Multipotent cell lineages in earlymouse development depend on SOX2 function. GenesDevelopment, 17, 126-140Behboodi E, Lam L, Gavin W G, Bondareva A, DobrinskiI 2013. Goat embryonic stem-like cell derivation andcharacterization. Methods in Mollecular Biology, 1074,51-67Le Bin G C, Munoz-Descalzo S, Kurowski A, Leitch H, Lou X,Mansfield W, Etienne-Dumeau C, Grabole N, Mulas C, NiwaH, Hadjantonakis A K, Nichols J. 2014. Oct4 is requiredfor lineage priming in the developing inner cell mass of themouse blastocyst. Development, 141, 1001-1010Boyer L A, Lee T I, Cole M F, Johnstone S E, Levine S S, ZuckerJ P, Guenther M G, Kumar R M, Murray H L, Jenner R G,Gifford D K, Melton D A, Jaenisch R, Young R A. 2005. Coretranscriptional regulatory circuitry in human embryonic stemcells. Cell, 122, 947-956De A K, Malakar D, Akshey Y S, Jena M K, Dutta R. 2011.Isolation and characterization of embryonic stem cell-likecells from in vitro produced goat (Capra hircus) embryos.Animal Biotechnology, 22, 181-196Dietrich J E, Hiiragi T. 2007. Stochastic patterning in the mousepre-implantation embryo. Development, 134, 4219-4231Du Puy L, Lopes S M, Haagsman H P, Roelen B A. 2011.Analysis of co-expression of OCT4, NANOG and SOX2 inpluripotent cells of the porcine embryo, in vivo and in vitro.Theriogenology, 75, 513-526Frankenberg S, Gerbe F, Bessonnard S, Belville C, PouchinP, Bardot O, Chazaud C. 2011. Primitive endodermdifferentiates via a three-step mechanism involving Nanogand RTK signaling. Developmental Cell, 21, 1005-1013Goentoro L, Shoval O, Kirschner M W, Alon U. 2009. Theincoherent feedforward loop can provide fold-changedetection in gene regulation. Molecular Cell, 36, 894-899Goissis M D, Cibelli J B. 2014. Functional characterizationof SOX2 in bovine preimplantation embryos. Biology ofReproduction, 90, 1-10Hambiliki F, Strom S, Zhang P, Stavreus-Evers A. 2012. Colocalizationof NANOG and OCT4 in human pre-implantationembryos and in human embryonic stem cells. Journal ofAssisted Reproduction and Genetics, 29, 1021-1028He S, Pant D, Schiffmacher A, Bischoff S, Melican D, Gavin W,Keefer C. 2006. Developmental expression of pluripotencydetermining factors in caprine embryos: Novel pattern ofNANOG protein localization in the nucleolus. MolecularReproduction and Development, 73, 1512-1522Keefer C L, Pant D, Blomberg L, Talbot N C. 2007. Challengesand prospects for the establishment of embryonic stemcell lines of domesticated ungulates. Animal ReproductionScience, 98, 147-168Keramari M, Razavi J, Ingman K A, Patsch C, Edenhofer F,Ward C M, Kimber S J. 2010. Sox2 is essential for formationof trophectoderm in the preimplantation embryo. PLoSOne, 5, e13952.Kirchhof N, Carnwath J W, Lemme E, Anastassiadis K,Scholer H, Niemann H. 2000. Expression pattern of Oct-4in preimplantation embryos of different species. Biology ofReproduction, 63, 1698-1705Kumar D, Anand T, Singh K P, Singh M K, Shah R A, ChauhanM S, Palta P, Singla S K, Manik R S. 2011. Derivation ofbuffalo embryonic stem-like cells from in vitro-producedblastocysts on homologous and heterologous feedercells. Journal of Assisted Reproduction and Genetics, 28,679-688Leeb M, Wutz A. 2011. Derivation of haploid embryonic stemcells from mouse embryos. Nature, 479, 131-134Li W, Shuai L, Wan H, Dong M, Wang M, Sang L, Feng C, LuoG Z, Li T, Li X, Wang L, Zheng Q Y, Sheng C, Wu H J, LiuZ, Liu L, Wang X J, Zhao X Y, Zhou Q. 2012. Androgenetichaploid embryonic stem cells produce live transgenic mice.Nature, 490, 407-411Messerschmidt D M, Kemler R. 2010. Nanog is required for primitive endoderm formation through a non-cellautonomous mechanism. Developmental Biology, 344,129-137Miyanari Y, Torres-Padilla M E. 2012. Control of ground-statepluripotency by allelic regulation of Nanog. Nature, 483,470-473Nichols J, Zevnik B, Anastassiadis K, Niwa H, Klewe-NebeniusD, Chambers I, Scholer H, Smith A. 1998. Formation ofpluripotent stem cells in the mammalian embryo dependson the POU transcription factor Oct4. Cell, 95, 379-391Ovitt C E, Scholer H R. 1998. The molecular biology of Oct-4 inthe early mouse embryo. Molecular Human Reproduction,4, 1021-1031Palmieri S L P W, Hess H, Schöler H R. 1994. Oct-4 transcriptionfactor is differentially expressed in the mouse embryoduring establishment of the first two extraembryonic celllineages involved in implantation. Developmental Biology,166, 259-267Pan H, Schultz R M. 2011. Sox2 modulates reprogrammingof gene expression in two-cell mouse embryos. Biology ofReproduction, 85, 409-416Pesce M, Scholer H R. 2001. Oct-4: gatekeeper in thebeginnings of mammalian development. Stem Cells, 19,271-278Singh R, Kumar K, Khan I, Ranjan R, T Y, Singh R K, DasB C, Bag S. 2013. Comparative expression profile ofdevelopmental related genes in haploid and diploidparthenogenetic embryos in caprine. The Indian Journalof Animal Sciences, 83, 42-45Tachibana M, Clepper L, Sparman M, Ramsey C, Mitalipov S.2009. The role of NANOG during primate preimplantationembryo development. Biology of Reproduction, 81, 248.Takeuchi T, Neri Q V, Katagiri Y, Rosenwaks Z, Palermo GD. 2005. Effect of treating induced mitochondrial damageon embryonic development and epigenesis. Biology ofReproduction, 72, 584-592Wu G, Han D, Gong Y, Sebastiano V, Gentile L, Singhal N,Adachi K, Fischedick G, Ortmeier C, Sinn M, Radstaak M,Tomilin A, Scholer H R. 2013. Establishment of totipotencydoes not depend on Oct4A. Nature Cell Biology, 15,1089-1097 |
| No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
