• Genomics & Informatics
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• v.20 no.2
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• pp.24.1-24.8
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• 2022

Hypomyelinating leukodystrophy type 2 (HLD2), is an inherited genetic disease of the central nervous system caused by recessive mutations in the gap junction protein gamma 2 (GJC2/GJA12). HLD2 is characterized by nystagmus, developmental delay, motor impairments, ataxia, severe speech problem, and hypomyelination in the brain. The GJC2 sequence encodes connexin 47 protein (Cx47). Connexins are a group of membrane proteins that oligomerize to construct gap junctions protein. In the present study, a novel missense mutation gene c.760G>A (p.Val254Met) was identified in a patient with HLD2 by performing whole exome sequencing. Following the discovery of the new mutation in the proband, we used Sanger sequencing to analyze his affected sibling and parents. Sanger sequencing verified homozygosity of the mutation in the proband and his affected sibling. The autosomal recessive inheritance pattern was confirmed since Sanger sequencing revealed both healthy parents were heterozygous for the mutation. PolyPhen2, SIFT, PROVEAN, and CADD were used to evaluate the function prediction scores of detected mutations. Cx47 is essential for oligodendrocyte function, including adequate myelination and myelin maintenance in humans. Novel mutation p.Val254Met is located in the second extracellular domain of Cx47, both extracellular loops are highly conserved and probably induce intramolecular disulfide interactions. This novel mutation in the Cx47 gene causes oligodendrocyte dysfunction and HLD2 disorder.

• Animal cells and systems
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• v.2 no.1
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• pp.99-106
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• 1998

In this study, we have examined the role of cAMP in gap junctional communication (GJC) in preimplantation mouse embryos. GJC was monitored by Lucifer Yellow (LY) injected into one blastomere of compacted embryos. The speed of GJC was defined as the time taken for the last blastomere of the embryo to become visibly fluorescent. The median time for 8-cell embrvos (140 sec) was similar to that for 16-cell (135 sec). To determine whether cAMP and cAMP-dependent protein kinase (PKA) are involved in the regulation of GJC, the effects of PKA inhibitor (H8) and cAMP analogues (Rp-cAMP and 8-Br-cAMP) on dye transfer between blastomeres of compacted embryos were examined. Some of the embryos treated with either H8 or Rp-cAMP failed to transfer LY to all blastomeres within 10 min. In contrast, 8-Br-cAMP speeded up fluorescent dye transfer. The median time to fill all blastomeres with LY was 140 sec in untreated controls and 90 sec in siblings treated with 8-Br-cAMP. Inhibition of PKA by H8 or Rp-cAMP induced delay or arrest in embryo development after compaction, but the increase of intracellular cAMP showed no effect. These findings suggest that GJC in preimplantation mouse embryos is regulated by cAMP-PKA pathway and transient interference by PKA inhibitors induces the developmental delay beyond compaction.

• Korean Journal of Ecology and Environment
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• v.45 no.1
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• pp.42-51
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• 2012

This study investigated the characteristics of natural organic matter (NOM) with general water characteristics (pH, DO, electrical conductivity, BOD, COD, TN, TP, Chl-$a$, DOC, $UV_{254}$, SUVA) and the 3D fluorescence excitation-emission matrix (FEEM) in the Yeongsan River basin. FEEM was used to classify protein-like and fulvic & humiclike substances with fluorescence intensity in the matrix of excitation and emission wavelength. The concentration of BOD, COD, TN, electrical conductivity and DOC in the region of Gwangju city (Gwangju sewage treatment plant: GJS, Gwangjucheon: GJC, Gwangju 2: GJ2) was relatively higher than the upper reaches and lower reaches of the Yeongsan River basin. SUVA in most sites was lower than 3 L $mg^{-1}\;m^{-1}$ as the hydrophilic substances, except Damyang (DY) in the upper reaches of Yeongsan river was higher than 3 L $mg^{-1}\;m^{-1}$ as the hydrophobic substances during winter and autumn. In the FEEM investigation the fulvic and humic substances were found in most sites, and in sites regarding Gwangju city (GJS, GJC, GJ2) during winter and GJC in summer, protein-like substances were found. The trend of fluorescence intensities from the upper reaches to the lower reaches in most sites corresponded to that regarding the concentration of water characteristics (BOD, COD, TN, DOC). That is why the region of Gwangju city (GJS, GJC, GJ2) was relatively higher. This results were an equivalent trend to those of fluorescence index (FI) in most sites, and the higher FIs in the sites of Gwangju city indicate more microbial-derived substances due to enormous effluent organic matters (EfOM) from huge Gwangju sewage treatment plants.

• Molecules and Cells
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• v.37 no.3
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• pp.270-274
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• 2014

Lymphatic vessels are essential to regulate interstitial fluid homeostasis and diverse immune responses. A number of crucial factors, such as VEGFC, SOX18, PROX1, FOX2C, and GJC2, have been implicated in differentiation and/or maintenance of lymphatic endothelial cells (LECs). In humans, dysregulation of these genes is known to cause lymphedema, a debilitating condition which adversely impacts the quality of life of affected individuals. However, there are no currently available pharmacological treatments for lymphedema, necessitating identification of additional factors modulating lymphatic development and function which can be targeted for therapy. In this report, we investigate the function of genes associated with Bone Morphogenetic Protein (BMP) signaling in lymphatic development using zebrafish embryos. The knock-down of BMP type II receptors, Bmpr2a and Bmpr2b, and type I receptors, Alk3 and Alk3b, as well as SMAD5, an essential cellular mediator of BMP signaling, led to distinct lymphatic defects in developing zebrafish. Therefore, it appears that each constituent of the BMP signaling pathway may have a unique function during lymphatic development. Taken together, our data demonstrate that BMP signaling is essential for normal lymphatic vessel development in zebrafish.

• Development and Reproduction
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• v.17 no.1
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• pp.63-72
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• 2013

A specific inhibitor of RNA polymerase II, ${\alpha}$-amanitin is broadly used to block transcriptional activities in cells. Previous studies showed that ${\alpha}$-amanitin affects in vitro maturation of cumulus-oocyte-complex (COC). In this study, we evaluated the target of ${\alpha}$-amanitin, and whether it affects oocytes or cumulus cells (CCs), or both. We treated ${\alpha}$-amanitin with different time period during in vitro culture of denuded oocytes (DOs) or COCs in comparison, and observed the changes in morphology and maturation status. Although DOs did not show any change in morphology and maturation rates with ${\alpha}$-amanitin treatment, oocytes from COCs were arrested at metaphase I (MI) stage and CCs were more scattered than control groups. To discover causes of meiotic arrest and scattering of CCs, we focused on changes of cumulus expansion, gap junctions, and cellular metabolism which to be the important factors for the successful in vitro maturation of COCs. Expression of genes for cumulus expansion markers (Ptx3, Has2, and Tnfaip6) and gap junctional proteins (Gja1, Gja4, and Gjc1) decreased in ${\alpha}$-amanitin-treated CCs. However, these changes were not observed in oocytes. In addition, expression of genes related to metabolism (Prps1, Rpe, Rpia, Taldo1, and Tkt) decreased in ${\alpha}$-amanitin-treated CCs but not in oocytes. Therefore, we concluded that the transcriptional activities of CCs for supporting suitable transcripts, especially for its metabolic activities and formation of gap junctions among CCs as well as with oocytes, are important for oocytes maturation in COCs.

• Korean Journal of Ecology and Environment
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• v.44 no.4
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• pp.358-363
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• 2011

This study investigated the characteristics of natural organic matter(NOM) with tXAD resin and FT-IR in the Yeongsan river system of Gwangju region. NOM fractionation by XAD 8/4 resins was used to classify hydrophobic and hydrophilic substances. FTIR was applied to classify functional groups in the structure of NOM. In the XAD investigation, most of the four site-samples were mainly hydrophilic substances. In March, hydrophobic substances were dominant in the Gwangju 1 site (GJ-1), while hydrophilic substances were dominant for the other sites. In May, samples of all four sites were hydrophilic with a vigorous activity of microorganism due to increasing temperatures. The October results were very similar with those from March. In the FT-IR investigation, most of the broad and large peaks were assigned to the aliphatic group, particularly the OH group, C-H, $C-H_2$, $C-H_3$, and C-O alcohol group. All were related to hydrophilic substances. Other peaks showed the aromatic group, particularly the C=O (Ketone) Group. As a result, there is an identification of NOM in the Yeongsan river system composing mainly of hydrophilic substances and functional groups (OH, C-H etc.) of the aliphatic compound.