• Genomics & Informatics
• /
• 제20권2호
• /
• pp.24.1-24.8
• /
• 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
• /
• 제2권1호
• /
• pp.99-106
• /
• 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.

• 생태와환경
• /
• 제45권1호
• /
• pp.42-51
• /
• 2012

본 연구는 영산강 수계의 일반적인 수질 특성(pH, DO, 전기전도도, BOD, COD, TN, TP, Chl-$a$, DOC, $UV_{254}$, SUVA)과 3D fluorescence excitation-emission matrix (FEEM) 분석을 통한 자연유기물질(NOM)의 특성을 조사하였다. FEEM은 여기 파장과 방출 파장에서의 형광세기를 이용하여 단백질계 (protein-like), 펄빅계 (fulvic-like) 및 휴믹계(humic-like) 물질을 분류하는데 사용된다. 일반수질 항목(BOD, COD, TN, 전기전도도 및 DOC)는 영산강 수계의 중류에 위치한 광주하수처리장(GJS), 광주천(GJC), 광주2 (GJ2) 등 광주광역시 지역에서 상류 및 하류 지역보다 상대적으로 높게 나타났다. 대부분의 지점에서 SUVA값은 3 L $mg^{-1}\;m^{-1}$보다 낮게 나타나 친수성 경향을 보이나, 영산강 상류지역인 담양(DY)에서는 겨울과 가을에 3 L $mg^{-1}\;m^{-1}$ 이상으로 소수성 경향을 보이고 있다. FEEM 조사결과, 대부분의 지점은 펄빅계와 휴믹계 물질이 검출되었으며, 다만 겨울에 광주지역(GJS, GJC, GJ2), 여름에 GJC 지점에서는 단백질계 물질이 검출되었다. 영산강 상류부터 하류지역까지 대부분의 지점에서 형광세기는 일반 수질항목(BOD, COD, TN, DOC) 농도와 유사한 경향을 보이고 있으며, 광주시 지역(GJS, GJC, GJ2)에서 상대적으로 높게 나타났다. 이 결과는 대부분의 지점에서 형광지표(Flourescence index, FI)의 경향과도 일치하며, 광주시에서 FI가 높게 나타난 것은 규모가 큰 광주시 하수처리장에서 방류되는 자연유기물질(EfOM)로 인한 미생물 기원 물질이 많기 때문으로 보여진다.

• Molecules and Cells
• /
• 제37권3호
• /
• pp.270-274
• /
• 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.

• 한국발생생물학회지:발생과생식
• /
• 제17권1호
• /
• pp.63-72
• /
• 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.

• 생태와환경
• /
• 제44권4호
• /
• pp.358-363
• /
• 2011

본 연구는 영산강수계 광주지역에 대하여 XAD와 FTIR를 이용하여 자연유기물질에 대한 분포특성을 연구하였다. XAD 8/4 resin을 이용한 NOM fractionation 분석은 친수성 및 소수성을 변별하는데 사용되어졌다. FT-IR은 자연유기물의 구조에서 기능족을 분석하는데 사용되어졌다. XAD 조사에서 영산강본류의 광주상류지점(GJ-1), 광주하류지점(GJ-2), 광주천(GJC), 광주하수처리장(GJS) 등 4개 지점에서 대부분 친수성으로 조사되었다. 다만, 3월에 GJ-1 지점에서는 소수성이 주를 이루었다. 5월에는 수온의 상승과 함께 미생물의 활성의 증가로 4개 지점 공통적으로 친수성으로 조사되었고, 10월에는 3월과 매우 유사한 결과가 되었다. FT-IR 조사에서는 넓거나 높아 뚜렷한 피크들이 주로 지방족 화합물, 특히 OH 그룹, CH, $C-H_2$, $C-H_3$ 및 C-O 알코올 그룹들로 나타났고, 이들은 친수성 물질들이다. FT-IR 조사에서 그 외의 피크들은 약간의 방향족 화합물, 특히 C=O (Ketone) 그룹이 나타났다. 결과적으로 본 연구에서 영산강수계 광주지역은 주로 친수성 물질들이며, 지방족 화합물(OH, C-H etc.)인 것으로 조사되었다.