• Clinical and Experimental Reproductive Medicine
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• 제41권2호
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• pp.47-61
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• 2014

Stored maternal factors in oocytes regulate oocyte differentiation into embryos during early embryonic development. Before zygotic gene activation (ZGA), these early embryos are mainly dependent on maternal factors for survival, such as macromolecules and subcellular organelles in oocytes. The genes encoding these essential maternal products are referred to as maternal effect genes (MEGs). MEGs accumulate maternal factors during oogenesis and enable ZGA, progression of early embryo development, and the initial establishment of embryonic cell lineages. Disruption of MEGs results in defective embryogenesis. Despite their important functions, only a few mammalian MEGs have been identified. In this review we summarize the roles of known MEGs in mouse fertility, with a particular emphasis on oocytes and early embryonic development. An increased knowledge of the working mechanism of MEGs could ultimately provide a means to regulate oocyte maturation and subsequent early embryonic development.

• 한국산학기술학회논문지
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• 제13권8호
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• pp.3532-3536
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• 2012

난자 세포의 정상적인 성숙과정을 이해하려면 모계유래 유전자 발현 증가의 분자 생물학적 기전을 밝혀내야 한다. 이것은 모계 유전자의 염기서열의 변화와 밀접한 관계가 있다. 전 연구결과에 의하면 돼지 난자 체외 성숙과정에서의 모계 유전자 mRNA 발현은 통상적으로 poly(A) 꼬리 길이와 아데닐산 중합반응에 의하여 검증된다. 하지만 포유동물 체외성숙 과정에서는 아직까지 밝혀진 것이 없다. 따라서 본 연구목적은 성숙단계 난모세포에서의 분자생물학적 기전을 해명하고자, 4개의 중요한 모계유전자발현을 real-time PCR기법으로 확인하여 poly(A) 꼬리 길이와 아데닐산중합반응의 변화를 확인하였다. 본 연구에서 접합체 유전자 활성화 단계에서 모계 유전자의 비정상적인 발현과 이것에 상응하는 단백질 수준의 억제는 일부 혹은 대부분 유전자 손실에 의하여 초래된 것임을 알 수 있었다. 따라서 이상적인 모계 유전자 발현은 난자 세포의 성숙 및 더 나가서 초기 배아 발달에 중요한 역할을 하는 것임을 확인 하였다.

• 한국수정란이식학회지
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• 제28권4호
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• pp.361-371
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• 2013

The present study assessed the effect of FSH and LH on oocyte meiotic, cytoplasmic maturation and on the expression level and polyadenylation status of several maternal genes. Cumulus-oocyte complexes were cultured in the presence of FSH, LH, or the combination of FSH and LH. Significant cumulus expansion and nuclear maturation was observed upon exposure to FSH alone and to the combination of FSH and LH. The combination of FSH and LH during entire IVM increased the mRNA level of four maternal genes, C-mos, Cyclin B1, Gdf9 and Bmp15, at 28 h. Supplemented with FSH or LH significantly enhanced the polyadenylation of Gdf9 and Bmp15; and altered the expression level of Gdf9 and Bmp15. Following parthenogenesis, the exposure of oocytes to combination of FSH and LH during IVM significantly increased cleavage rate, blastocyst formation rate and total cell number, and decreased apoptosis. In addition, FSH and LH down-regulated the autophagy gene Atg6 and upregulated the apoptosis gene Bcl-xL at the mRNA level in blastocysts. These data suggest that the FSH and LH enhance meiotic and cytoplasmic maturation, possibly through the regulation of maternal gene expression and polyadenylation. Overall, we show here that FSH and LH inhibit apoptosis and autophagy and improve parthenogenetic embryo competence and development.

• Journal of Genetic Medicine
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• 제12권2호
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• pp.100-108
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• 2015

Purpose: Conventional methods for the prenatal detection of fetal RhD status involve invasive procedures such as fetal blood sampling and amniocentesis. The identification of cell-free fetal DNA (cffDNA) in maternal plasma creates the possibility of determining fetal RhD status by analyzing maternal plasma DNA. However, some technical problems still exist, especially the lack of a positive control marker for the presence of fetal DNA. Therefore, we assessed the feasibility and accuracy of fetal RHD genotyping incorporating the RASSF1A epigenetic fetal DNA marker from cffDNA in the maternal plasma of RhD-negative pregnant women in Korea. Materials and Methods: We analyzed maternal plasma from 41 pregnant women identified as RhD-negative by serological testing. Multiplex real-time PCR was performed by amplifying RHD exons 5 and 7 and the SRY gene, with RASSF1A being used as a gender-independent fetal epigenetic marker. The results were compared with those obtained by postnatal serological analysis of cord blood and gender identification. Results: Among the 41 fetuses, 37 were RhD-positive and 4 were RhD-negative according to the serological analysis of cord blood. There was 100% concordance between fetal RHD genotyping and serological cord blood results. Detection of the RASSF1A gene verified the presence of cffDNA, and the fetal SRY status was correctly detected in all 41 cases. Conclusion: Noninvasive fetal RHD genotyping with cffDNA incorporating RASSF1A is a feasible, reliable, and accurate method of determining fetal RhD status. It is an alternative to amniocentesis for the management of RhD-negative women and reduces the need for unnecessary RhIG prophylaxis.

• Journal of Preventive Medicine and Public Health
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• 제42권1호
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• pp.1-4
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• 2009

Objectives : Isolated cleft lip with or without cleft palate(CL/P) is among the most common human birth defects, with a prevalence of approximately 1 in 700 live births. The B-Cell Leukemia/lymphoma 3(BCL3) gene has been suggested as a candidate gene for CL/P based on association and linkage studies in some populations. This study tests for an association between markers in BCL3 and isolated, non-syndromic CL/P using a case-parent trio design, while considering parent-of-origin effects. Methods : Forty case-parent trios were genotyped for two single nucleotide polymorphisms(SNPs) in the BCL3 gene. We performed a transmission disequilibrium test(TDT) on individual SNPs, and the FAMHAP package was used to estimate haplotype frequencies and to test for excess transmission of multi-SNP haplotypes. Results : The odds ratio for transmission of the minor allele, OR(transmission), was significant for SNP rs8100239(OR=3.50, p=0.004) and rs2965169(OR=2.08, p=0.027) when parent-of-origin was not considered. Parentspecific TDT revealed that SNP rs8100239 showed excess maternal transmission. Analysis of haplotypes of rs2965169 and rs8100239 also suggested excess maternal transmission. Conclusions : BCL3 appears to influence risk of CL/P through a parent-of-origin effect with excess maternal transmission.

• Asian-Australasian Journal of Animal Sciences
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• 제17권9호
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• pp.1192-1196
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• 2004

In order to detect the molecular mechanism of heterosis in pigs, the mRNA differential display technique was performed to investigate the differences in gene expression of pig's Longissimus dorsi between the high-parent heterosis cross combination Landrace${\times}$Large White and the mid-parent heterosis cross combination Large White${\times}$Meishan. Three pig purebreds, Large White, Meishan, and Landrace and four types of reciprocal $F_1$ hybrids were analyzed using nine 3'-end anchored primers in combination with ten 5'-end arbitrary primers and nearly 7,000 reproducible bands were examined. The patterns of gene expression of each cross combination were analyzed and eight common patterns (fifteen kinds) were found. When the results from the two cross combinations were put together and compared, eight different typical expression patterns were observed, these indicated that the patterns of gene expression of these two cross combinations had obvious differences. Gene expression correlation and cluster analyses of the two cross combinations indicated that the gene expression of the mid-parent heterosis cross combination was correlated with maternal effect, but in the high-parent heterosis cross combination, paternal effect acted in the gene expression of the hybrids or the gene expression of the hybrids was biased towards one parent.

• Asian-Australasian Journal of Animal Sciences
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• 제27권1호
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• pp.19-23
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• 2014

Here, we studied the genetic diversity of native fowls in Laos by analyzing a mitochondrial DNA (mtDNA) sequence polymorphism. A 546-bp fragment of the mtDNA D-loop region was sequenced in 129 chickens from the areas of Vientiane, Luang Prabang and Pakse. In total, 29 haplotypes were identified and formed five clades. Haplotype diversity and nucleotide diversity of the native fowls in Laos were $0.85536{\pm}0.0172$ and $0.010158{\pm}0.005555$, respectively. Although the Laotian native fowls were distributed across five clades, most of them were clustered in two main clades (A and B), which were originated in China. The other haplotypes were contained in clades D, F, and I, which originated from continental southeast Asia. These results suggest that multiple maternal lineages were involved in the origin of domestic chicken in Laos. Moreover, there appear to be at least two maternal lineages, one from China and the other from the southeast Asian continent.

• Journal of Genetic Medicine
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• 제13권2호
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• pp.65-71
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• 2016

Cell-free nucleic acids (cf-NAs) originate in trophoblasts and are detected in the maternal plasma. Using innovative bioinformatic technologies such as next-generation sequencing, cf-NAs in the maternal plasma have been rapidly applied in prenatal genetic screening for fetal aneuploidy. Amniotic fluid is a complex and dynamic fluid that provides growth factors and protection to the fetus. In 2001, the presence of cf-NA in amniotic fluid was reported. Amniotic fluid is in direct contact with the fetus and is derived from fetal urine and maternal and fetal plasma. Therefore, these genetic materials have been suggested to reflect fetal health and provide real-time genetic information regarding fetal development. Recently, several studies evaluated the global gene expression changes of amniotic fluid cell-free RNA according to gestational age. In addition, by analyzing the transcriptome in the amniotic fluid of fetal aneuploidy, potential key pathways and novel biomarkers for fetal chromosomal aneuploidy were identified. Here, we review the current knowledge of cell-free RNA in amniotic fluid and suggest future research directions.

• 한국임상수의학회지
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• 제33권5호
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• pp.266-269
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• 2016

Cell-free fetal RNA is useful to determine fetal sex and detect other inherent genetic disorders. However, non-invasive fetal sex determination methods using fetal RNA from maternal plasma is not yet well established in studies pertaining to bovine animals. Thus, the aim of this study was to systematically evaluate the presence of the male-specific ZRSR2Y gene transcript in maternal plasma using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) assays, and to verify its accuracy, sensitivity, and specificity in determining fetal sex between 30 and 100 days of gestation. Overall accuracy, sensitivity, and specificity of the ZRSR2Y gene transcripts in determining fetal sex were 89.1%, 86.3%, and 100%, respectively. The 30 to 100 days of gestation were further classified into five stages of gestation, and each stage had relatively high accurate, sensitive, and specific results. Overall, these results indicate that the expression of the ZRSR2Y gene can be used for fetal sex determination in bovine animals using circulating cell-free RNA in maternal plasma during early pregnancy.

• 한국가축번식학회지
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• 제23권4호
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• pp.365-369
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• 1999

Interferon tau (IFN$\tau$), the pregnancy recognition signal in ruminants, suppresses transcription of the estrogen receptor (ER) gene in the endometrial luminal (LE) and superficial glandular epithelium (sGE) to prevent oxytocin receptor (OTR) expression and pulsatile release of luteolytic prostaglandin $F_{2{\alpha}}$ (PGF), Interferon regulatory factors one (IRF-l) and two (IRF-2) are transcription factors induced by IFN$\tau$ that activate and silence gene expression, respectively. Available results suggest that IFN$\tau$ acts directly on LE and sGE during pregnancy to induce sequentially IRF-l and then IRF-2 gene expression to silence transcription of ER and OTR genes, block the luteolytic mechanism to maintenance a functional corpus luteum (CL) and, signal maternal recognition of pregnancy. The theory for maternal recognition of pregnancy in pigs is that the uterine endometrium of cyclic gilts secretes PGF in an endocrine direction, toward the uterine vasculature for transport to the CL to exert its luteolytic effect. However, in pregnant pigs, estrogens secreted by the conceptuses are responsible, perhaps in concert with effects of prolactin and calcium, for exocrine secretion of PGF into the uterine lumen where it is sequestered to exert biological effects and / or be metabolized to prevent luteolysis.