Browse > Article

Methylation Status of H19 Gene in Embryos Produced by Nuclear Transfer of Spermatogonial Stem Cells in Pig  

Lee, Hyun-Seung (Department of Animal Biotechnology, Animal Resources Research Center/Bio-Organ Research Center, Konkuk University)
Lee, Sung-Ho (Department of Animal Biotechnology, Animal Resources Research Center/Bio-Organ Research Center, Konkuk University)
Gupta, Mukesh Kumar (Department of Animal Biotechnology, Animal Resources Research Center/Bio-Organ Research Center, Konkuk University)
Uhm, Sang-Jun (Department of Animal Biotechnology, Animal Resources Research Center/Bio-Organ Research Center, Konkuk University)
Lee, Hoon-Taek (Department of Animal Biotechnology, Animal Resources Research Center/Bio-Organ Research Center, Konkuk University)
Publication Information
Abstract
The faulty regulation of imprinting gene lead to the abnormal development of reconstructed embryo after nuclear transfer. However, the correlation between the imprinting status of donor cell and preimplantation stage of embryo development is not yet clear. In this study, to determine this correlation, we used the porcine spermatogonial stem cell (pSSC) and fetal fibroblast (pFF) as donor cells. As the results, the isolated cells with laminin matrix selection strongly expressed the GFR ${\alpha}$-1 and PLZF genes of SSCs specific markers. The pSSCs were maintained to 12 passages and positive for the pluripotent marker including OCT4, SSEA1 and NANOG. The methylation analysis of H19 DMR of pSSCs revealed that the zinc finger protein binding sites CTCF3 of H19 DMRs displayed an androgenic imprinting pattern (92.7%). Also, to investigate the reprogramming potential of pSSCs as donor cell, we compared the development rate and methylation status of H19 gene between the reconstructed embryos from pFF and pSSC. This result showed no significant differences of the development rate between the pFFs ($11.2{\pm}0.8%$) and SSCs ($13.3{\pm}1.1%$). However, interestingly, while the CTCF3 methylation status of pFF-NT blastocyst was decreased (36.3%), and the CTCF3 methylation status of pSSC-NT blastocyst was maintained. Therefore, this result suggested that the genomic imprinting status of pSSCs is more effective than that of normal somatic cells for the normal development because the maintenance of imprinting pattern is very important in early embryo stage.
Keywords
Porcine SSCs; bFGF; Laminin; Reprogramming; SCNT; Imprinting gene;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Uhm SJ, Gupta MK, Das ZC, Kim JH, Park C, Kim T, Lee HT (2009): Effect of transgene introduction and recloning on efficiency of porcine transgenic cloned embryo production in vitro. Reprod Domest Anim 44:106-115.   DOI   ScienceOn
2 Wakayama T, Yanagimachi R (1999): Cloning of male mice from adult tail-tip cells. Nat Genet 22:127-128.   DOI   ScienceOn
3 Wilmut I, Schnieke AE, McWhir J, Kind AJ, Campbell KH (1997): Viable offspring derived from fetal and adult mammalian cells. Nature 385:810-813.   DOI   ScienceOn
4 Zovoilis A, Nolte J, Drusenheimer N, Zechner U, Hada H, Guan K, Hasenfuss G, Nayernia K, Engel W (2008): Multipotent adult germline stem cells and embryonic stem cells have similar microRNA profiles. Mol Hum Reprod 14:521-529.   DOI   ScienceOn
5 Shinohara T, Avarbock MR, Brinster RL (1999): Betal- and alpha 6-integrin are surface markers on mouse spermatogonial stem cells. Proc Natl Acad Sci 96:5504-5509.   DOI   ScienceOn
6 Shinohara T, Orwig KE, A varbock MR, Brinster RL (2002): Germ line stem cell competition in postnatal mouse testes. BioI Reprod 66:1491-1497.   DOI   ScienceOn
7 Meng X, Lindahl M, Hyvonen ME, Parvinen M, de Rooij DG, Hess MW, Raatikainen-Ahokas A, Sainio K, Rauvala H, Lakso M, Pichel JG, Westphal H, Saarma M, Sariola H (2000): Regulation of cell fate decision of undifferentiated spermatogonia by GDNF. Science 287:1489-1493.   DOI   ScienceOn
8 Stoger R, Kubicka P, Liu CG, Kafri T, Razin A, Cedar H, Barlow DP (1993): Maternal-specific methylation of the imprinted mouse Igf2r locus identifies the expressed locus as carrying the imprinting signal. Cell 73:61-71.   DOI   ScienceOn
9 Surani MA, Allen ND, Barton SC, Fundele R, Howlett SK, Norris ML, Reik W (1990): Developmental consequences of imprinting of parental chromosomes by DNA methylation. Philos Trans R Soc Lond B BioI Sci 326:313-327.   DOI
10 Takagi Y, Talbot NC, Rexroad CE Jr, Pursel VG (1997): Identification of pig primordial germ cells by immunocytochemistry and lectin binding. Mol Reprod Dev 46:567-580.   DOI   ScienceOn
11 Mizukami T, Kanai Y, Fujisawa M, Kanai-Azuma M, Kurohmaru M, Hayashi Y (2008): Five azacytidine, a DNA methyltransferase inhibitor, specifically inhibits testicular cord formation and Sertoli cell differentiation in vitro. Mol Reprod Dev 75:1002-1010.   DOI   ScienceOn
12 Oh SH, Jung YH, Gupta MK, Uhm SJ, Lee HT (2009): H19 gene is epigenetically stable in mouse multipotent germline stem cells. Mol Cells 27:635-640.   DOI   ScienceOn
13 Reik W, Dean W, Walter J (2001): Epigenetic reprogramming in mammalian development. Science 293:1089-1093.   DOI   ScienceOn
14 Rideout WM, Wakayama T, Wutz A, Eggan K, Jackson-Grusby L, Dausman J, Yanagimachi R, Jaenisch R (2000): Generation of mice from wild-type and targeted ES cells by nuclear cloning. Nat Genet 24:109-110.   DOI   ScienceOn
15 Tegelenbosch RA, de Rooij DG (1993): A quantitative study of spermatogonial multiplication and stem cell renewal in the C3H/101 $F_1$ hybrid mouse. Mutat Res 290:193-200.   DOI   ScienceOn
16 Tremblay KD, Saam JR, Ingram RS, Tilghman SM, Bartolomei MS (1995): A paternal-specific methylation imprint marks the alleles of the mouse H19 gene. Nat Genet 9:407-413.   DOI   ScienceOn
17 Rideout WM, Eggan K, Jaenisch R (2001): Nuclear cloning and epigenetic reprogramming of the genome. Science 293:1093-1098.   DOI
18 Roh S, Choi HY, Park SK, Won C, Kim BW, Kim JH, Kang H, Lee ER, Cho SG (2009): Porcine nuclear transfer using somatic donor cells altered to express male germ cell function. Reprod Fertil Dev 21:882-891.   DOI   ScienceOn
19 Ju JY, Park CY, Gupta MK, Uhm SJ, Paik EC, Ryoo ZY, Cho YH, Chung KS, Lee HT (2008): Establishment of stem cell lines from nuclear transferred and parthenogenetically activated mouse oocytes for therapeutic cloning. Fertil Steril 89:1314-1323.   DOI   ScienceOn
20 Mayer W, Niveleau A, Walter J, Fundele R, Haaf T (2000): Demethylation of the zygotic paternal genome. Nature 403:501-502.
21 Kuijk EW, Colenbrander B, Roelen BA (2009): The effects of growth factors on in vitro-cultured porcine testicular cells. Reproduction 138:721-731.   DOI   ScienceOn
22 Latham KE (1999): Mechanisms and control of embryonic genome activation in mammalian embryos. Int Rev Cytol 193:71-124.
23 Lucifero D, Mertineit C, Clarke HJ, Bestor TH, Trasler JM (2002): Methylation dynamics of imprinted genes in mouse germ cells. Genomics 79:530-538.   DOI   ScienceOn
24 Jaenisch R (2002): Nuclear cloning, embryonic stem cells, and transplantation therapy. Harvey Lect 98:145-171.
25 Luo J, Megee S, Rathi R, Dobrinski I (2006): Protein gene product 9.5 is a spermatogonia-specific marker in the pig testis: application to enrichment and culture of porcine spermatogonia. Mol Reprod Dev 73:1531-1540.   DOI   ScienceOn
26 Han DW, Im YB, Do JT, Gupta MK, Uhm SJ, Kim JH, Scholer HR, Lee HT (2008): Methylation status of putative differentially methylated regions of porcine IGF2 and H19. Mol Reprod Dev 75:777-784.   DOI   ScienceOn
27 Izadyar F, Spierenberg GT, Creemers LB, den Ouden K, de Rooij DG (2002): Isolation and purification of type A spermatogonia from the bovine testis. Reproduction 124:85-94.   DOI   ScienceOn
28 Goel S, Sugimoto M, Minami N, Yamada M, Kume S, Imai H (2007): Identification, isolation, and in vitro culture of porcine gonocytes. BioI Reprod 77:127-137.   DOI   ScienceOn
29 Gupta MK, Uhm SJ, Lee HT (2008): Sexual maturity and reproductive phase of oocyte donor influence the developmental ability and apoptosis of cloned and parthenogenetic porcine embryos. Anim Reprod Sci 108:107-121.   DOI
30 Eggan K, Rode A, Jentsch I, Samuel C, Hennek T, Tintrup H, Zevnik B, Erwin J, Loring J, Jackson-Grusby L, Speicher MR, Kuehn R, Jaenisch R (2002): Male and female mice derived from the same embryonic stem cell clone by tetraploid embryo complementation. Nat Biotechnol 20:455-459.   DOI   ScienceOn
31 Brinster RL (2002): Germline stem cell transplantation and transgenesis. Science 296:2174-2176.   DOI   ScienceOn
32 Guan K, Nayernia K, Maier LS, Wagner S, Dressel R, Lee JH, Nolte J, Wolf F, Li M, Engel W, Hasenfuss G (2006): Pluripotency of spermatogonial stem cells from adult mouse testis. Nature 440:1199-1203.   DOI   ScienceOn
33 Gupta MK, Uhm SJ, Lee HT (2007): Differential but beneficial effect of phytohemagglutinin on efficiency of in vitro porcine embryo production by somatic cell nuclear transfer or in vitro fertilization. Mol Reprod Dev 74:1557-1567.   DOI   ScienceOn
34 Cattanach BM, Beechey CV(1990): Autosomal and X-chromosome imprinting. Dev Suppl pp 63-72.
35 Costoya JA, Hobbs RM, Barna M, Cattoretti G, Manova K, Sukhwani M, Orwig KE, Wolgemuth DJ, Pandolfi PP (2004): Essential role of Plzf in maintenance of spermatogonial stem cells. Nat Genet 36:653-659.
36 Dirami G, Ravindranath N, Pursel V, Dym M (1999): Effects of stem cell factor and granulocyte macrophage-colony stimulating factor on survival of porcine type A spermatogonia cultured in KSOM. BioI Reprod 61:225-230.   DOI   ScienceOn
37 Carr MS, Yevtodiyenko A, Schmidt CL, Schmidt JV (2007): Allele-specific histone modifications regulate expression of the Dlk1-Gtl2 imprinted domain. Genomics 89:280-290.   DOI   ScienceOn