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Loss of Imprinting of IGF2 Gene in the Chorionic Tissues of Spontaneously Eliminated Human Embryos.

Zastavna D, Makukh H, Tretjak B, Bilevych O, Tyrka M - Genet Epigenet (2013)

Bottom Line: Genomic imprinting in the chorionic tissues of spontaneously eliminated human embryos has been studied on the model of 820-AG (Apa1) of the IGF-2 gene locus.Molecular and genetic analysis was performed on the polymorphic 820-AG IGF2 locus in 107 samples of DNA extracted from the chorionic tissues of spontaneously eliminated human embryos within 5-10 weeks of gestation.Presence of AG genotype Apa1 single nucleotide polymorphisms of the IGF-2 was shown to cause more than a 7-fold increase in the risk of embryo elimination.

View Article: PubMed Central - PubMed

Affiliation: Institute of Hereditary Pathology NAMS of Ukraine, Lviv.

ABSTRACT
Insulin-like growth factor-2 (IGF-2) is a mitogen, growth and differentiation modulator for many cell types. It is mainly expressed during the prenatal development, and its activity strongly depends on the genomic imprinting. Genomic imprinting in the chorionic tissues of spontaneously eliminated human embryos has been studied on the model of 820-AG (Apa1) of the IGF-2 gene locus. Molecular and genetic analysis was performed on the polymorphic 820-AG IGF2 locus in 107 samples of DNA extracted from the chorionic tissues of spontaneously eliminated human embryos within 5-10 weeks of gestation. Presence of AG genotype Apa1 single nucleotide polymorphisms of the IGF-2 was shown to cause more than a 7-fold increase in the risk of embryo elimination. Thus, the loss of genomic imprinting of the IGF-2 gene may be an important cause of the miscarriages in human.

No MeSH data available.


Related in: MedlinePlus

Electrophoresis in 2% agarose gel of Apa1 restriction fragments of exon 9 of the IGF-2 gene: 1–100 bp DNA Ladder; 2, 3, 4-820AG genotype; 5, 6-820GG genotype; 7-820AA genotype.
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f1-geg-5-2013-017: Electrophoresis in 2% agarose gel of Apa1 restriction fragments of exon 9 of the IGF-2 gene: 1–100 bp DNA Ladder; 2, 3, 4-820AG genotype; 5, 6-820GG genotype; 7-820AA genotype.

Mentions: Isolation and purification of DNA from the chorionic villi cells were performed using enzymatic cleavage of tissue with subsequent phenol extraction. Amplification of DNA sequences was performed by polymerase chain reaction (PCR) in the automatic mode in thermal cycler “Tertsyk” (“DNA Technology”, Russian Federation). Restriction endonuclease, thermostable Taq-polymerase and oligonucleotide primers (“Fermentas”, Vilnius, Lithuania) were used as described by Tadokoro et al.8 Electrophoretic separation of products of PCR-RFLP analysis (Restriction Fragments Length Polymorphism) was conducted in 2% agarose gels that were further stained with the ethidium bromide and scanned on UV trans-illuminator.3 The 292 bp fragment was amplified by the PCR and treated with Apa1 endonuclease (37 °C, 16 hours [h]). If the restriction site was absent, the amplified product was not changed and provided the 292 bp fragment (820AA genotype). In cases where the 820GG IGF-2 genotype appeared in the DNA sample, the 229 bp and 63 bp fragments were observed. In cases where the heterozygous 820AG genotype was present, three fragments of 292, 229 and 63 bp appeared on the electropherogram (Fig. 1).


Loss of Imprinting of IGF2 Gene in the Chorionic Tissues of Spontaneously Eliminated Human Embryos.

Zastavna D, Makukh H, Tretjak B, Bilevych O, Tyrka M - Genet Epigenet (2013)

Electrophoresis in 2% agarose gel of Apa1 restriction fragments of exon 9 of the IGF-2 gene: 1–100 bp DNA Ladder; 2, 3, 4-820AG genotype; 5, 6-820GG genotype; 7-820AA genotype.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4222333&req=5

f1-geg-5-2013-017: Electrophoresis in 2% agarose gel of Apa1 restriction fragments of exon 9 of the IGF-2 gene: 1–100 bp DNA Ladder; 2, 3, 4-820AG genotype; 5, 6-820GG genotype; 7-820AA genotype.
Mentions: Isolation and purification of DNA from the chorionic villi cells were performed using enzymatic cleavage of tissue with subsequent phenol extraction. Amplification of DNA sequences was performed by polymerase chain reaction (PCR) in the automatic mode in thermal cycler “Tertsyk” (“DNA Technology”, Russian Federation). Restriction endonuclease, thermostable Taq-polymerase and oligonucleotide primers (“Fermentas”, Vilnius, Lithuania) were used as described by Tadokoro et al.8 Electrophoretic separation of products of PCR-RFLP analysis (Restriction Fragments Length Polymorphism) was conducted in 2% agarose gels that were further stained with the ethidium bromide and scanned on UV trans-illuminator.3 The 292 bp fragment was amplified by the PCR and treated with Apa1 endonuclease (37 °C, 16 hours [h]). If the restriction site was absent, the amplified product was not changed and provided the 292 bp fragment (820AA genotype). In cases where the 820GG IGF-2 genotype appeared in the DNA sample, the 229 bp and 63 bp fragments were observed. In cases where the heterozygous 820AG genotype was present, three fragments of 292, 229 and 63 bp appeared on the electropherogram (Fig. 1).

Bottom Line: Genomic imprinting in the chorionic tissues of spontaneously eliminated human embryos has been studied on the model of 820-AG (Apa1) of the IGF-2 gene locus.Molecular and genetic analysis was performed on the polymorphic 820-AG IGF2 locus in 107 samples of DNA extracted from the chorionic tissues of spontaneously eliminated human embryos within 5-10 weeks of gestation.Presence of AG genotype Apa1 single nucleotide polymorphisms of the IGF-2 was shown to cause more than a 7-fold increase in the risk of embryo elimination.

View Article: PubMed Central - PubMed

Affiliation: Institute of Hereditary Pathology NAMS of Ukraine, Lviv.

ABSTRACT
Insulin-like growth factor-2 (IGF-2) is a mitogen, growth and differentiation modulator for many cell types. It is mainly expressed during the prenatal development, and its activity strongly depends on the genomic imprinting. Genomic imprinting in the chorionic tissues of spontaneously eliminated human embryos has been studied on the model of 820-AG (Apa1) of the IGF-2 gene locus. Molecular and genetic analysis was performed on the polymorphic 820-AG IGF2 locus in 107 samples of DNA extracted from the chorionic tissues of spontaneously eliminated human embryos within 5-10 weeks of gestation. Presence of AG genotype Apa1 single nucleotide polymorphisms of the IGF-2 was shown to cause more than a 7-fold increase in the risk of embryo elimination. Thus, the loss of genomic imprinting of the IGF-2 gene may be an important cause of the miscarriages in human.

No MeSH data available.


Related in: MedlinePlus