Restricted accessResearch articleFirst published online 2016-10
Effect of Sex of Embryo on Developmental Competence,Epigenetic Status,and Gene Expression in Buffalo ( Bubalus bubalis ) Embryos Produced by Hand-Made Cloning
Buffalo embryos were produced by hand-made cloning using skin fibroblasts from male and female buffaloes (n = 4 each) as donor cells for examining the effect of sex. Although the rate of blastocyst formation (43.8% ± 1.31% vs. 42.2% ± 1.22%) was similar, the total cell number (333 ± 10.4 vs. 270 ± 10.9) was higher (p < 0.05) whereas the apoptotic index (6.39 ± 0.25 vs. 8.52 ± 0.38) was lower (p < 0.05) for male than for female blastocysts. In the blastocysts, the global level of H3K18ac was found to be in the following order: male>female>IVF (in vitro fertilization) blastocysts (p < 0.05). The global level of H3K9me2 was not significantly different between male and female blastocysts and was higher (p < 0.05) compared with that in their IVF counterparts. The relative mRNA abundance of X-chromosome-linked (XIST, HPRT, PGK, and G6PD), apoptosis- (CASPASE3) and pregnancy-related genes (IFN-τ) was significantly higher (p < 0.05) whereas that of DNMT1 was significantly lower (p < 0.05) in female than in male blastocysts; however, in the case of apoptosis- (BCL-XL) and developmental competence-related genes (IGF1R and OCT4), the expression level was similar between the two groups. The gene expression level of OCT4 and IFN-τ but not of IGF1R was significantly lower (p < 0.05) in cloned than in IVF blastocysts. This study demonstrates that the epigenetic status, quality, and expression level of several genes but not the developmental competence are affected by the sex of cloned embryos.
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References
1.
AmarnathD., LiX.P., KatoY., and TsunodaY. (2007). Gene expression in individual bovine somatic cell cloned embryos at 8-cell and blastocyst stages. J. Reprod. Dev., 53, 1247–1263.
2.
BazerF., SpencerT., and OttT. (1997). Interferon tau: A novel pregnancy recognition signal. Am. J. Reprod. Immunol., 37, 412–420.
3.
Bermejo-AlvarezP., LonerganP., RathD., and Gutierrez-AdanA. (2010a). Developmental kinetics and gene expression in male and female bovine embryos produced in vitro with sex-sorted spermatozoa. Reprod. Fertil. Dev., 22, 426–436.
4.
Bermejo-AlvarezP., RizosD., RathD., LonerganP., and Gutierrez-AdanA. (2010b). Sex determines the expression level of one third of the actively expressed genes in bovine blastocysts. Proc. Nat. Acad. Sci. U. S. A., 107, 3394–3399.
5.
BestorT.H. (2000). The DNA methyltransferases of mammals. Hum. Mol. Genet., 9, 2395–2402.
6.
BeyhanZ., ForsbergE.J., EilertsenK.J., Kent-FirstM., and FirstN.L. (2007). Gene expression in bovine nuclear transfer embryos in relation to donor cell efficiency in producing live offspring. Mol. Reprod. Dev., 74, 18–27.
7.
Chavatte-PalmerP., CamousS., JammesH., Le Cleac'hN., GuillomotM., and LeeR.S. (2012). Review: Placental perturbations induce the developmental abnormalities often observed in bovine somatic cell nuclear transfer. Placenta, 33Suppl: S99–S104.
8.
CuiX.S., XuY.N., ShenX.H., ZhangL.Q., ZhangJ.B., and KimN.H. (2011). Trichostatin A modulates apoptotic-related gene expression and improves embryo viability in cloned bovine embryos. Cell. Reprogram., 13, 179–189.
9.
EgganK., AkutsuH., HochedlingerK., RideoutW.3rd, YanagimachiR., and JaenischR. (2000). X-Chromosome inactivation in cloned mouse embryos. Science, 290, 1578–1581.
10.
FeldmanN., GersonA., FangJ., LiE., ZhangY., ShinkaiY., CedarH., and BergmanY. (2006). G9a-mediated irreversible epigenetic inactivation of Oct-3/4 during early embryogenesis. Nat. Cell Biol., 8, 188–194.
11.
GalliC., LagutinaI., PerotaA., ColleoniS., DuchiR., LucchiniF., and LazzariG. (2012). Somatic cell nuclear transfer and transgenesis in large animals: Current and future insights. Reprod. Domest. Anim., 47(Suppl 3), 2–11.
12.
Geng-ShengC., YuG., KunW., Fang-RongD., and NingL. (2009). Repressive but not activating epigenetic modifications are aberrant on the inactive X chromosome in live cloned cattle. Dev. Growth Differ., 51, 585–594.
13.
Gutierrez-AdanA., BehboodiE., AndersonG.B., MedranoJ.F., and MurrayJ.D. (1996). Relationship between stage of development and sex of bovine IVM-IVF embryos cultured in vitro versus in the sheep oviduct. Theriogenology, 46, 515–525.
14.
Gutierrez-AdanA., OterM., Martinez-MadridB., PintadoB., and De LaFuenteJ. (2000). Differential expression of two genes located on the X chromosome between male and female in vitro-produced bovine embryos at the blastocyst stage. Mol. Reprod. Dev., 55, 146–151.
15.
HembergerM. (2002). The role of the X chromosome in mammalian extra-embryonic development. Cytogenet. Genome Res., 99, 210–217.
16.
InoueK., KohdaT., SugimotoM., SadoT., OgonukiN., MatobaS., ShiuraH., IkedaR., MochidaK., FujiiT., SawaiK., OtteA.P., TianX.C., YangX., IshinoF., AbeK., and OguraA. (2010). Impeding Xist expression from the active X chromosome improves mouse somatic cell nuclear transfer. Science, 330, 496–499.
17.
JonesP.L., VeenstraG.J.C., WadeP.A., VermaakD., KassS.U., LandsbergerN., StrouboulisJ., and WolffeA.P. (1998). Methylated DNA and MeCP2 recruit histone deacetylase to repress transcription. Nat. Genet., 19, 187–191.
18.
KangY.K., KooD.B., ParkJ.S., ChoiY.H., ChungA.S., LeeK.K., and HanY.M. (2001). Aberrant methylation of donorgenome in cloned bovine embryos. Nat. Genet., 28, 173–177.
19.
KobayashiS., IsotaniA., MiseN., YamamotoM., FujiharaY., KasedaK., NakanishiT., IkawaM., HamadaH., AbeK., and OkabeM. (2006). Comparison of gene expression in male and female mouse blastocysts revealed imprinting of the X-linked gene, Rhox5/Pem, at preimplantation stages. Curr. Biol., 16, 166–172.
20.
LarsonM.A., KimuraK., KubischH.M., and RobertsR.M. (2001). Sexual dimorphism among bovine embryos in their ability to make the transition to expanded blastocyst and in the expression of the signaling molecule IFN-tau. Proc. Natl. Acad. Sci. U. S. A., 98, 9677–9682.
21.
LathamK.E., PatelB., BautistaF.D., and HawesS.M. (2000). Effects of X chromosome number and parental origin on X-linked gene expression in preimplantation mouse embryos. Biol. Reprod., 63, 64–73.
22.
LeeD.Y., HayesJ.J., PrussD., and WolffeA.P. (1993). A positive role for histone acetylation in transcription factor access to nucleosomal DNA. Cell, 72, 73–84.
23.
LoiP., ModlinskiaJ.A., and PtakJ. (2011). Interspecies somatic cell nuclear transfer: A salvage tool seeking first aid. Theriogenology, 76, 217–228.
24.
LyonM.F. (1961). Gene action in the X-chromosome of the mouse (Musmusculus L.). Nature, 190, 372–373.
25.
MatobaS., InoueK., KohdaT., SugimotoM., MizutaniE., OgonukI.N., NakamuraT., AbeK., NakanoT., IshinoF., and OguraA. (2011). RNAi-mediated knockdown of Xist can rescue the impaired postimplantation development of cloned mouse embryos. Proc. Natl. Acad. Sci. U. S. A., 108, 20621–20626.
26.
MohapatraS.K., SandhuA., NeerukattuV.S., SinghK.P., SelokarN.L., SinglaS.K., ChauhanM.S., ManikR.S., and PaltaP. (2015a). Buffalo embryos produced by handmade cloning from oocytes selected using BCB staining have better developmental competence and quality, and are closer to embryos produced by in vitro fertilization in terms of their epigenetic status and gene expression pattern. Cell. Reprogram., 17, 141–150.
27.
MohapatraS.K., SandhuA., SinghK.P., SinglaS.K., ChauhanM.S., ManikR.S., and PaltaP. (2015b). Establishment of trophectoderm cell lines from buffalo (Bubalus bubalis) embryos of different sources and examination of in vitro developmental competence, quality, epigenetic status and gene expression in cloned embryos derived from them. PLoS One, 10, e0129235.
28.
NiemannH., TianX.C., KingW.A., and LeeR.S. (2008). Epigenetic reprogramming in embryonic and foetal development upon somatic cell nuclear transfer cloning. Reproduction, 135, 151–163.
29.
NolenL.D., GaoS., HanZ., MannM.R., Gie ChungY., OtteA.P., BartolomeiM.S., and LathamK.E. (2005). X chromosome reactivation and regulation in cloned embryos. Dev. Biol., 279, 525–540.
30.
ParkC.H., JeongY.H., JeongY.I., LeeS.Y., JeongY.W., ShinT., KimN.H., JeungE.B., HyunS.H., LeeC.K., LeeE., and HwangW.S. (2012). X-linked gene transcription patterns in female and male in vivo, in vitro and cloned porcine individual blastocysts. PLoS One, 7, e51398.
31.
PeippoJ., FarazmandA., KurkilahtiM., MarkkulaM., BasrurP.K., and KingW.A. (2002). Sex-chromosome linked gene expression in in-vitro produced bovine embryos. Mol. Hum. Reprod., 8, 923–929.
32.
RiceJ.C., and AllisC.D. (2001). Histone methylation versus histone acetylation: New insights into epigenetic regulation. Curr. Opin. Cell. Biol., 13, 263–273.
33.
SantosF., ZakhartchenkoV., StojkovicM., PetersA., JenuweinT., WolfE., ReikW., and DeanW. (2003). Epigenetic marking correlates with developmental potential in cloned bovine preimplantation embryos. Curr. Biol., 13, 1116–1121.
34.
SelokarN.L., SainiM., MuzaffarM., KrishnakanthG., SahaA.P., ChauhanM.S., ManikR.S., PaltaP., MadanP., and SinglaS.K. (2012a). Roscovitine treatment improves synchronization of donor cell cycle in G0/G1 stage and in vitro development of handmade cloned buffalo (Bubalus bubalis) embryos. Cell. Reprogram., 14, 146–154.
35.
SelokarN.L., SainiM., PaltaP., ChauhanM.S., ManikR.S., and SinglaS.K. (2014). Hope for restoration of dead valuable bulls through cloning using donor somatic cells isolated from cryopreserved semen. PLoS One, 9:e90755.
36.
SelokarN.L., ShahR.A., SahaA.P., MuzaffarM., SainiM., ChauhanM.S., ManikR.S., PaltaP., and SinglaS.K. (2012b). Effect of post-fusion holding time, orientation and position of somatic cell-cytoplasts during electrofusion on the development of handmade cloned embryos in buffalo (Bubalus bubalis). Theriogenology, 78, 930–936.
37.
SharmaR., GeorgeA., KambleN.M., SinghK.P., ChauhanM.S., SinglaS.K., ManikR.S., and PaltaP. (2011). Optimization of culture conditions to support long-term self-renewal of buffalo (Bubalus bubalis) embryonic stem cell-like cells. Cell. Reprogram., 13, 539–549.
38.
WrenzyckiC., Lucas-HahnA., HerrmannD., LemmeE., KorsaweK., and NiemannN. (2002). In vitro production and nuclear transfer affect dosage compensation of the X-linked gene transcripts G6PD, PGK, and Xist in preimplantation bovine embryos. Biol. Reprod., 66, 127–134.
39.
XueF., TianX.C., DuF., KubotaC., TanejaM., DinnyesA., DaiY., LevineH., PereiraL.V., and YangX. (2002). Aberrant patterns of X chromosome inactivation in bovine clones. Nat. Genet., 31, 216–220.
40.
YamanakaK., SugimuraS., WakaiT., KawaharaM., and SatoE. (2009). Acetylation level of histone H3 in early embryonic stages affects subsequent development of miniature pig somatic cell nuclear transfer embryos. J. Reprod. Dev., 55, 638–644.
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