Aberrant expression of miR‐29a/29b and methylation level of mouse embryos after in vitro fertilization and vitrification at two‐cell stage

2019 ◽  
Vol 234 (10) ◽  
pp. 18942-18950 ◽  
Author(s):  
Elham Movahed ◽  
Mansooreh Soleimani ◽  
Sara Hosseini ◽  
Azade Akbari Sene ◽  
Mohammad Salehi
Keyword(s):  
In Vitro Fertilization ◽  
Methylation Level ◽  
Mouse Embryos ◽  
Cell Stage ◽  
Aberrant Expression ◽  
Vitro Fertilization

scholarly journals Male advantage observed for in vitro fertilization mouse embryos exhibiting early cleavage

2020 ◽  
Author(s):  
Yosuke Kawase ◽  
Takanori Tachibe ◽  
Nobuo Kamada ◽  
Kou‐ichi Jishage ◽  
Hiroyuki Watanabe ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Mouse Embryos ◽  
Early Cleavage ◽  
Vitro Fertilization ◽  
Male Advantage

285 THE EFFICIENCY OF INTRACYTOPLASMIC SPERM INJECTION v. LASER-ASSISTED IN VITRO FERTILIZATION WITH FROZEN SPERM FOR RECOVERY TRANSGENIC C57BL/6 MOUSE STRAINS

2011 ◽  
Vol 23 (1) ◽  
pp. 240
Author(s):  
A. C. Carstea ◽  
Z. Polgar ◽  
L. Kovacs ◽  
A. Dinnyes
Keyword(s):  
In Vitro Fertilization ◽  
Intracytoplasmic Sperm Injection ◽  
Skim Milk ◽  
Mouse Strains ◽  
Chi Square ◽  
Cell Stage ◽  
Important Indicator ◽  
Vitro Fertilization ◽  
Cryopreserved Sperm

The progress of molecular genetics generated thousands of new transgenic strains of mice which also requires their economic and safe maintenance in the form of genetic banks. Sperm freezing would be one of the easiest options; however, cryosensitivity of sperm in mice strains is prone to variation. In this study, we examined the efficiency of laser-assisted zona drilling in vitro fertilization (ZD-IVF) v. intracytoplasmic sperm injection (ICSI) for attempting to recover two transgenic (UBI-GFP/BL6 and B6;129P2- Hvcn1) and one mutant (C57BL/6J-Tyrc-2J) lines. The sperm was frozen with 18% raffinose and 3% skim milk (Nakagata 2000 Mamm Genome 11, 572–576). Data of the replicates was analysed by chi-square method. The motility rates after thawing of cryopreserved sperm were 10% for UBI-GFP/BL6, 30% C57BL/6J-Tyrc-2J and 50% for B6;129P2-Hvcn1 strains. Regular IVF attempts in the UBI-GFP/BL6 and the mutant strain resulted in very few embryos and no pups (data not shown). Following ZD-IVF, the 2-cell stage rates were 7/60 (12%) for UBI-GFP/BL6, 18/60 (30%) for C57BL/6J-Tyrc-2J, and 34/60 (56%) for B6;129P2- Hvcn1. After ICSI, 66–74% of the oocytes survived the procedure and their development to 2-cells stage were 12/20 (60%) (UBI-GFP/BL6), 25/39 (64%) (C57BL/6J-Tyrc-2J) and 26/37 (70%) (B6;129P2-Hvcn1). The number of 2-cell stage embryos produced by ICSI was significantly (P < 0.05) increased compared with those produced following ZD-IVF in case of UBI-GFP/BL6 and C57BL/6J-Tyrc-2J strains. The 2-cell stage embryos were transferred into recipients and the newborn rates from ZD-IVF v. ICSI embryos were 0% v. 17% (UBI-GFP/BL6), 28% v. 36% (C57BL/6J-Tyrc-2J) and 12% v. 12% (B6;129P2- Hvcn1), respectively; none of them were significantly different. In conclusion, when using cryopreserved sperm, the post-thaw motility is an important indicator for the selection of the rederivation method of cryopreserved transgenic mouse strains; while ZD-IVF, an easier method to perform, is suitable for the higher motility samples, ICSI could be strongly recommended for those showing low motility. This work was financed by EU FP6: CLONET (MRTN-CT-2006-035468), TEAMOHOLIC (MEXT-CT-2003-509582); EU FP7: RESOLVE (FP7-HEALTH-F4-2008-202047), RabPStem (PERG07-GA-2010-268422), and NKFP_07_1-ES2HEART-HU (OM-00202-2007).

Development of androgenetic mouse embryos produced by in vitro fertilization of enucleated oocytes

1993 ◽  
Vol 34 (1) ◽  
pp. 43-46 ◽  
Author(s):  
Tomohiro Kono ◽  
Yusuke Sotomaru ◽  
Yasuhiro Sato ◽  
Tatsuo Nakahara
Keyword(s):  
In Vitro Fertilization ◽  
Mouse Embryos ◽  
Vitro Fertilization

50 DNA METHYLATION PROFILES OF UPSTREAM ELEMENTS OF Oct-3/4 GENE IN IN VITRO FERTILIZATION (IVF) AND SOMATIC CELL NUCLEAR-TRANSFERRED (SCNT) EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 143
Author(s):  
M. Kawasumi ◽  
Y. Unno ◽  
M. Nishiwaki ◽  
K. Matsumoto ◽  
M. Anzai ◽  
...  
Keyword(s):  
Dna Methylation ◽  
In Vitro Fertilization ◽  
Somatic Cell ◽  
Methylation Level ◽  
Nuclear Transfer ◽  
Upstream Region ◽  
Sequencing Analysis ◽  
Vitro Fertilization ◽  
Cpg Sites

Cloning by adult somatic cell nuclear transfer (SCNT) has proven to be successful for the production of clones from many species (Keith 2004 Cytogenet. Genome Res. 105, 285). However, somatic cloning is currently highly inefficient. One of the reasons for this is that SCNT is believed to be associated with epigenetic errors including abnormal DNA methylation of the reconstructed embryo. The Oct-3/4 gene, a member of the POU transcription factor family, is expressed throughout the pre-implantation embryo. Abnormal expression of the Oct-3/4 gene in the nuclear-transferred embryo is either directly or indirectly caused by nuclear transfer and is suggested to be indicative of a general failure to reset the genetic program (Boiani et al. 2002 Genes Dev. 16, 1209). In this study, we investigated the DNA methylation profiles of the Oct-3/4 gene in the genome of SCNT embryos, using bisulfite sequencing analysis. Then, we observed the detailed subcellular localization of Oct-3/4 proteins in SCNT embryos using immunocytochemical (ICC) analysis. Nuclear transfer of cumulus cell nuclei was carried out as previously described (Wakayama et al. 1998 Nature 394, 369). After nuclear transfer, embryos were subsequently cultured in KSOM media to the morula and blastocyst stages. We compared the methylation profiles of 3 transcriptional control elements (distal enhancer, DE; proximal enhancer, PE; and promoter) of the upstream region of the Oct-3/4 gene with the genome of in vitro fertilization (IVF) and SCNT embryos. The methylation rate of CpG sites in the DE and promoter regions of both IVF and SCNT embryos was low at both the morula and the blastocyst stages. What&apos;s interesting is that there was a significant difference in the methylation level on CpG sites in the PE element between IVF and SCNT embryos. At the morula stage, the methylation level on CpG sites in the PE element was very low in the IVF embryo and moderately high in the SCNT embryo (0.9&percnt; and 26.3&percnt;). Conversely, at the blastocyst stage, CpG sites in the PE element showed high methylation in the IVF embryo and low methylation in the SCNT embryo (55.2&percnt; and 10.5&percnt;). CpG sites in the PE element were lightly methylated (3.0&percnt;) in the inner cell mass (ICM) of the IVF embryo. This means that the main portion of methylation in the IVF blastocyst embryo occurred at the trophectoderm (TE). On the other hand, in ICM of the SCNT embryo, the methylation level of each embryonic cell was almost the same in the whole blastocyst embryo (9.8&percnt; and 10.5&percnt;). As a result, it is highly possible that the CpG sites in the PE element of ICM were methylated as in the TE. ICC analysis revealed that some SCNT embryos showed aberrant Oct-3/4 expression in the TE. These results indicate that the methylation of CpG sites in the Oct-3/4 PE element may be related to expression of Oct-3/4 in the mouse IVF and SCNT embryos. These differences in methylation level between IVF and SCNT embryos were reflected as abnormal expressions of Oct-3/4 on SCNT embryos. This study was supported by the 21st COE Program of MEST. M.K. is a JSPS Research Fellow and supported by Grant-in Aid for Scientific Research (No. 1751132) of JSPS.

scholarly journals Determining factors of early embryo cleavage to 2-cell stage for predicting the pregnancy outcome of in vitro fertilization

2003 ◽  
Vol 80 ◽  
pp. 153 ◽  
Author(s):  
Bo Sun Joo ◽  
Hwa Sook Moon ◽  
Sae Hee Park ◽  
Soo Kyung Lee ◽  
Myung Sun Lee ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Pregnancy Outcome ◽  
Early Embryo ◽  
Cell Stage ◽  
Vitro Fertilization ◽  
Determining Factors ◽  
Embryo Cleavage

Does in vitro fertilization affect the expression of miRNAs and their biogenesis pathway in preimplantation mouse embryos?

2019 ◽  
Vol 112 (1) ◽  
pp. 62-70
Author(s):  
Elham Azizi ◽  
Marefat Ghaffari Novin ◽  
Mohammad Naji ◽  
Fardin Amidi ◽  
Zahra Shams Mofarahe
Keyword(s):  
In Vitro Fertilization ◽  
Mouse Embryos ◽  
Vitro Fertilization ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse
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