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Comparative proteomic analysis associated with term placental insufficiency in cloned pig
Lee, So-Young,Park, Jong-Yi,Choi, Yun-Jung,Cho, Seong-Keun,Ahn, Jong Deok,Kwon, Deug-Nam,Hwang, Kyu-Chan,Kang, Sung-Jo,Paik, Seung-Sam,Seo, Han Geuk,Lee, Hoon Taek,Kim, Jin-Hoi WILEY-VCH 2007 PROTEOMICS -WEINHEIM- Vol.7 No.8
<P>Somatic cell-derived nuclear transfer (scNT) is a method of animal cloning in which the oocyte reprograms a somatic cell nucleus to divide and execute developmental programs. Despite many successes in this field, cloning by scNT remains very inefficient. Unlike other cloned animals, pigs derived by scNT have placentas with severe villous hypoplasia. To obtain a better understanding of the protein networks involved in this phenomenon, we assessed global protein expression profiles in term placentas from scNT-derived and control animals. Proteomic analysis of term placentas from scNT-derived animals identified 43 proteins that were differentially expressed compared to control animals. Among them, 14-3-3 proteins and Annexin V, which are closely involved in the apoptotic signaling pathway, were significantly down- and up-regulated, respectively. Western blot analysis and immunohistochemistry indicated that down-regulation of 14-3-3 proteins in scNT-derived placentas induced apoptosis of cytotrophoblast cells via mitochondria-mediated apoptosis. Taken together, our results suggest that placental insufficiency in scNT-derived placentas may be due to apoptosis, induced in part by the down-regulation of 14-3-3 proteins and up-regulation of Annexin V. They also indicate that proteomic maps represent an important tool for future studies of placental insufficiency and pathology.</P>
Lee, JoonHo,Kim, Chong Jai,Kim, Jung‐,Sun,Lee, Deug‐,Chan,Ahn, Sejin,Yoon, Bo Hyun John Wiley and Sons Inc. 2018 JOURNAL OF CELLULAR AND MOLECULAR MEDICINE Vol.22 No.2
<P><B>Abstract</B></P><P>Acute chorioamnionitis, frequently observed in preterm placentas, is a major risk factor for the development of infection and non‐infection‐related adverse perinatal outcomes. MicroRNAs play important roles in immune cell development and function as well as in the development of cancers and neurologic diseases. We sought to investigate the changes in microRNA‐223 (miR‐223) expression and the functional significance of the changes in miR‐223 expression in foetal organs in the presence of acute chorioamnionitis. Using formalin‐fixed, paraffin‐embedded (FFPE) tissue samples from foetal or neonatal autopsy cases, which are the most practical option to study the changes in several organs simultaneously, miR‐223 expression profiles in foetal thymus, lung and liver were compared between cases with and without acute chorioamnionitis. Total RNA was extracted from FFPE specimens and qRT‐PCR was conducted. miR‐223‐3p expression levels in foetal thymus (2.55‐fold), lung (1.93‐fold) and liver (1.70‐fold) were significantly higher in cases with acute chorioamnionitis than in those without. Transfection of pre‐miR‐223‐3p in Jurkat cells and luciferase assay and ribonucleoprotein immunoprecipitation followed by qRT‐PCR analysis confirmed the binding of miR‐223 to the 3′ untranslated region (3′UTR) of forkhead box O1 (FoxO1) mRNA and the regulation of FoxO1 by miR‐223. We report for the first time that foetuses with inflammation in the chorioamniotic membranes show increased expression of miR‐223 in the thymus, lung and liver. Furthermore, FoxO1 is a target of miR‐223. These findings suggest that post‐transcriptional regulation of genes by miR‐223 is a component of the foetal inflammatory response, which has systemic consequences in the foetus.</P>
Depigmentation of skin and hair color in the somatic cell cloned pig
Hwang, Kyu-Chan,Cho, Seong-Keun,Lee, Seong-Hoon,Park, Jong-Yi,Kwon, Deug-Nam,Choi, Yun-Jung,Park, Chankyu,Kim, Jae-Hwan,Park, Keun-Kyu,Hwang, Seongsoo,Park, Soo-Bong,Kim, Jin-Hoi Wiley-Liss, Inc. 2009 Developmental dynamics Vol.238 No.7
<P>Previously, we have successfully produced nine cloned piglets using Duroc donor cells. Among these clones, one showed distinct depigmentation of the skin and hair color during puberty. In this study, we selected a clone with depigmentation to investigate the etiology of the anomaly in somatic cell nuclear transfer. We hypothesized that genes related to Waardenburg syndrome (Mitf, Pax-3, Sox-10, Slug, and Kit) are closely associated with the depigmentation of pig, which was derived from somatic cell nuclear transfer (scNT). Total RNA was extracted from the ear tissue of affected and unaffected scNT-derived pigs, and the transcripts encoding Mitf, Pax-3, Sox-10, and Slug, together with the Kit gene, were amplified by reverse transcription-polymerase chain reaction, sequenced, and analyzed. The cDNA sequences from the scNT pig that showed progressive depigmentation did not reveal a mutation in these genes. Although we did not find any mutations in these genes, expression of the genes implicated in Waardenburg syndrome was severely down-regulated in the affected scNT pig when compared with unaffected scNT pigs. This down-regulation of gene expression may result in a previously undescribed phenotype that shows melanocyte instability, leading to progressive loss of pigmentation. Developmental Dynamics 238:1701–1708, 2009. © 2009 Wiley-Liss, Inc.</P>
Chang-Kyu Kim,Deug-Chan Lee,Suk-Ho Choi 한국축산식품학회 2017 한국축산식품학회지 Vol.37 No.4
Korean native honey (KNH) is much more expensive than European honey (EH) in Korea, because KNH is a favored honey which is produced less than EH. Food fraud of KNH has drawn attention of the government office concerned, which is in need of a method to differentiate between KNH and EH which are produced by the Asiatic honeybee, Apis cerana and the European honeybee, Apis mellifera, respectively. A method to discriminate KNH and EH was established by using duplex polymerase chain reaction (PCR) in this study. Immunochromatographic assay (IC) was examined to analyze the duplex PCR product. The DNA sequences of primers for the duplex PCR were determined by comparing cytochrome C oxidase genes of the two honey bee species. Chelex resin method was more efficient in extracting genomic DNA from honey than the other two procedures of commercial kits. The duplex PCR amplifying DNA of 133 bp were more sensitive than that amplifying DNA of 206 bp in detecting EH in the honey mixture of KNH and EH. Agarose gel electrophoresis and IC detected the DNA of 133 bp at the ratios of down to 1% and 5% EH in the honey mixture, respectively and also revealed that several KNH products distributed by internet shopping sites were actually EH. In conclusion, the duplex PCR with subsequent IC could also discriminate between KNH and EH and save time and labor.