바위솔속 엽육조직 세포 내 액포의 미세구조 분화 양상

김인선,Kim, In-Sun 한국현미경학회 2009 Applied microscopy Vol.39 No.4

다육질성 CAM 식물에서는 구조와 기능의 분화가 환경조건에 잘 적응된 합리적인 광합성을 수행하여 동일한 엽육세포에서 $CO_2$ 고정, 유기물 합성과 저장, 분해 및 활용하는 시간이 서로 다르게 나타난다. 이러한 유기산 대사는 CAM 식물의 가장 뚜렷한 대사적 특징으로 밤에 말산을 합성하여 액포에 저장하고 낮에 이용하므로 이들의 액포는 급격한 pH의 차이를 일주기성으로 조절해야 하는 매우 중요한 세포소기관이다. 본 연구에서는 식물체 내 생리적 건조가 지속되어 CAM 광합성을 수행하는 바위솔속 식물 3종의 다육질성 엽육조직 세포의 특성을 액포 구조분화에 초점을 두어 미세구조적으로 연구하였다. 바위솔속의 다육질성 엽육조직은 수분저장성 세포들로 구성되어 있으며, 액포융합 등의 액포화현상과 액포 내 다양한 2차 액포형성이 현저한 구조적 특징이었다. 이들 액포는 매우 역동적이어서 분열하여 다수의 소액포를 형성하거나 소액포들의 융합으로 큰 액포를 형성하였고, 일부는 전자밀도가 높은 저장성 액포로 발달하였다. 이러한 액포화는 세포의 크기를 경제적이고 에너지 효율적으로 증가시키는 방식으로 대부분의 다육질성 CAM 식물에서 발달하며, 낮과 밤에 일주기성으로 반복되는 세포 내 pH 농도의 급격한 변화를 대처할 수 있게 한다. 또한, 막 함입에 의한 다양한 크기의 수많은 2차 액포 형성은 단 기간 내에 액포막의 용적을 증가시켜 이러한 목적을 충족시켜 주는데 일주기적으로 사용되는 매우 중요한 세포 내 구획이 된다. 액포의 신장으로 세포질은 세포벽 주변부위로 밀려나 얇은 층으로 국한되었으나, 이들 세포질 내에서도 엽록체와 미토콘드리아는 액포와 밀접하게 연관되어 분포하고, 세포 간에는 원형질연락사가 잘 발달하였다. 이러한 미세구조들의 발달은 다육질성 엽육세포가 일주기성으로 급변하는 세포 내 유기산 대사과정에 적응하기 위해 액포에서의 신속하고 원활한 대사물질의 수송이 이루어져야 하기 때문일 것으로 추정된다. In the present study, ultrastructural features of the mesophyll tissue have been investigated in Crassulacean acid metabolism (CAM)-performing succulent Orostachys. A large central vacuole and numerous small vacuoles in the peripheral cytoplasm were characterized at the subcellular level in both developing and mature mesophyll cells. The most notable feature was the invagination of vacuolar membranes into the secondary vacuoles or multivesicular bodies. In many cases, tens of single, membrane-bound secondary vacuoles of various sizes were found to be formed within the central vacuole. multivesicular bodies containing numerous small vesicles were also distributed in the cytoplasm but were better developed within the central vacuole. Occasionally, electron-dense prevacuolar compartments, directly attached to structures appearing to be small vacuoles, were also detected in the cytoplasm. One or more huge central vacuoles were frequently observed in cells undergoing differentiation and maturation. Consistent with the known occurrence of morphologically distinct vacuoles within different tissues, two types of vacuoles, one representing lytic vacuoles and the other, most likely protein storage vacuoles, were noted frequently within Orostachys mesophyll. The two types coexisted in mature vegetative cells but did not merge during the study. Nevertheless, the coexistence of two distinct vacuole types in maturing cells implies the presence of more than one mechanism for vacuolar solute sorting in these species. The vacuolar membrane is known to be unique among the intracellular compartments for having different channels and/or pumps to maintain its function. In CAM plants, the vacuole is a very important organelle that regulates malic acid diurnal fluctuation to a large extent. The membrane invagination seen in Orostachys mesophyll likely plays a significant role in survival under the physiological drought conditions in which these Orostachys occur; by increasing to such a large vacuolar volume, the mesophyll cells are able to retain enormous amounts of acid when needed. Furthermore, the mesophyll cells are able to attain their large sizes with less energy expenditure in order to regulate the large degree of diurnal fluctuation of organic acid that occurs within the vacuoles of Orostachys.

Distal Myopathy with Rimmed Vacuoles Confirmed by Whole Exome Sequencing

Seong Don Seo(서승돈),Hyung Jun Park(박형준),Hyun Seok Song(송현석),Hye Jin Kim(김혜진),Jin-Mo Park(박진모),Young Bin Hong(홍영빈),Ki Wha Chung(정기화),Byung-Ok Choi(최병옥) 한국생명과학회 2014 생명과학회지 Vol.24 No.3

Rimed vacuole을 가진 원위 근육병(distal myopathy with rimmed vacuoles, DMRV)은 제2형 유전성 봉입체 근육병으로도 불리며 초기 성인기에 발병하여 원위부의 근력약화를 보이는 임상양상과 rimmed vacuole의 근육병리소견을 특징으로 하는 상염색체 열성의 근육병이다. 이러한 DMRV의 원인은 UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE) 유전자의 돌연변이임이 밝혀져 있다. 저자들은 원위부 근력약화를 호소 하는 환자에서 전체 엑솜 염기서열분석을 이용하여 GNE 유전자의 복합 이형접합성 돌연변이(p.Asp176Val 및 p.Val572Leu)를 확인하여 DMRV를 진단할 수 있었다. 본 연구는 근육병의 정확한 분자진단에 있어서 전체 엑솜 염기서열분석의 유용성을 보여주었기에 이를 보고하는 바이다. Distal myopathy with rimmed vacuoles (DMRV) or hereditary inclusion body myopathy 2 is an autosomal recessive muscular disorder characterized by early adult-onset weakness of distal muscles and rimmed vacuoles in muscle biopsy. Mutations in the UDP-N-acetylglucosamine 2-epimerase/N-ace-tylmannosamine kinase (GNE) gene are associated with the development of DMRV. In this study, whole exome sequencing (WES) revealed compound heterozygous GNE mutations of p.Asp176Val and p.Val572Leu in a patient with distal limb weakness. Three hundred healthy controls did not show these mutations. All other variants found in distal myopathy-relevant genes were polymorphic. These findings confirmed that the patient had DMRV. This work underscores the usefulness of WES in improving the molecular diagnosis of myopathy.

6-Aminonicotinamide 투여 후 햄스터 척수의 미세구조 변화

양영철,Yang, Young-Chul 한국현미경학회 1997 Applied microscopy Vol.27 No.3

The effects of antimetabolite, 6-aminonicotinamide (6-AN), on ultrastrudural changes in the spinal cord of golden hamster were investigated. Intraperitoneal administration of 6-AN (10 mg/kg body weight) every two days gave rise to a marked reduction of about $30\sim40%$ in body weight after $26\sim28$ days ($13\sim14th$ injection). In the lesions of the spinal cord, neuroglial cells such as astrocytes and oligodendrocytes were severely damaged, but neurons and blood vessels were not affected by 6-AN. The myelin sheath was also affected by 6-AN. Vacuoles observed in the lesions were produced by the swelling and degenerating changes of neuropils and neuroglial cells. Numerous swollen mitochondria and cisterns of rough endoplasmic reticulum were observed in the watery cytoplasm of damaged neuroglial cells, but intermediate filaments were well preserved. Especially in the damaged astrocytes, the outer nuclear membrane were partially swollen and formed a halfmoonlike structure. It is suggested that as well as the multivesicular bodies protruding from the swollen dendrites, the conjugation of adjacent vacuoles also participated in the formation of large vacuoles.

Detection and discrimination of <i>Shigella sonnei</i> and <i>Shigella flexneri</i> based on vacuolar responses in <i>Saccharomyces cerevisiae</i>

Nguyen, Ngoc-Tu,Park, Ra-Mi,Kim, Yang-Hoon,Min, Jiho Elsevier 2018 Journal of biotechnology Vol.287 No.-

<P><B>Abstract</B></P> <P>This study provided a system for bacteria detection based on a lysosome-like-vacuole response in the yeast <I>Saccharomyces cerevisiae</I>. Vacuoles are factors known to activate the immune system in the presence of foreign substances. Here, <I>Shigella sonnei</I> and <I>Shigella flexneri</I> were exposed to yeast to analyze the alteration of vacuolar enzymes. The ability to detect the bacteria was evaluated by confocal microscopy after exposing and staining vacuoles with LysoTracker. Results showed that the treatment of yeast with these bacteria increased the number of red vacuole-like organelles surrounding yeast nuclei. Thus, vacuole alteration can be used as a biomarker for bacteria detection. Next, the expression of vacuolar enzymes under the influence of bacteria was examined using two-dimensional gel electrophoresis (2-DE) method for screening specific biomarkers for each <I>Shigella</I> strain. Finally, the recombinant yeasts that contained biomarkers fused to different fluorescent proteins confirmed the ability of yeast to detect these two <I>Shigella</I> strains at concentrations ranging from 10 to 100 CFU/mL.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Detection of <I>Shigella</I> strains was performed using vacuolar response in <I>S. cerevisiae</I>. </LI> <LI> The vacuolar proteomic in response to <I>Shigella</I> was analyzed for screening of specific biomarkers. </LI> <LI> Biomarker for each strain was fused with different fluorescent proteins for the distinct detection. </LI> <LI> The ability of recombinant yeasts to recognize <I>Shigella</I> strains individually from 10 CFU/ml. </LI> </UL> </P>

Effect of heavy metals, pesticides and pharmaceuticals on yeast’s vacuoles as a biomarker for toxic detection

NGUYENNGOC TU,Simranjeet Singh Sekhon,윤지희,Yang-Hoon Kim,Jiho Min,Y.-H. Kim 대한독성 유전단백체 학회 2017 Molecular & cellular toxicology Vol.13 No.3

In this study, the effect of different toxic chemicals on Saccharomyces cerevisiae’s vacuoles was determined. More precisely, oxidative stress was mediated by pesticides and heavy metals, whereas nonoxidative stress influence was caused by the treatment with pharmaceuticals. Previously, oxidative stress was reported to alter yeast vacuolar enzymes, thus, intracellular vacuoles are assumed to be used as a biomarker to detect the oxidative stresses on yeast. Simultaneously, nonoxidative stress effect was expected to be recognized by vacuoles as well. The feasibility of toxic detection by vacuolar response was evaluated by confocal microscopy after staining the vacuoles with LysoTracker. The results indicate that each chemical has its own optimal concentration at which the signal corresponding to vacuolar response reached the peak without inhibiting the yeast growth. Therefore, this method was confirmed to detect toxic chemicals at their sub-lethal doses. In addition, the minimum response time of vacuoles for different toxic chemicals was also measured.

Bioengineered yeast-derived vacuoles with enhanced tissue-penetrating ability for targeted cancer therapy

Gujrati, Vipul,Lee, Miriam,Ko, Young-Joon,Lee, Sangeun,Kim, Daejin,Kim, Hyungjun,Kang, Sukmo,Lee, Soyoung,Kim, Jinjoo,Jeon, Hyungsu,Kim, Sun Chang,Jun, Youngsoo,Jon, Sangyong National Academy of Sciences 2016 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF Vol.113 No.3

<P>Despite the appreciable success of synthetic nanomaterials for targeted cancer therapy in preclinical studies, technical challenges involving their large-scale, cost-effective production and intrinsic toxicity associated with the materials, as well as their inability to penetrate tumor tissues deeply, limit their clinical translation. Here, we describe biologically derived nanocarriers developed from a bioengineered yeast strain that may overcome such impediments. The budding yeast Saccharomyces cerevisiae was genetically engineered to produce nanosized vacuoles displaying human epidermal growth factor receptor 2 (HER2)-specific affibody for active targeting. These nanosized vacuoles efficiently loaded the anticancer drug doxorubicin (Dox) and were effectively endocytosed by cultured cancer cells. Their cancer-targeting ability, along with their unique endomembrane compositions, significantly enhanced drug penetration in multicellular cultures and improved drug distribution in a tumor xenograft. Furthermore, Dox-loaded vacuoles successfully prevented tumor growth without eliciting any prolonged immune responses. The current study provides a platform technology for generating cancer-specific, tissue-penetrating, safe, and scalable biological nanoparticles for targeted cancer therapy.</P>

6-Aminonicotinamide 투여 후 햄스터 뇌의 형태변화

양영철,조병필,황태선,강호석 연세대학교 원주의과대학 1997 원주의대논문집 Vol.10 No.1

Effects of in vitro Yeast Vacuoles on 3T3-L1 Adipocyte Development and Differentiation

Suyeon CHOI,Yang-Hoon KIM,Jiho MIN 한국생물공학회 2020 한국생물공학회 학술대회 Vol.2020 No.10

Effects of in vitro Yeast Vacuoles on Adipocytes, 3T3-L1 Development and Differentiation

Suyeon CHOI,Jiho MIN 한국생물공학회 2021 한국생물공학회 학술대회 Vol.2021 No.4

Bioengineered Yeast-derived Vacuoles with Enhanced Tissue-Penetrating Ability for Targeted Cancer Therapy

Vipul GUJRATI,Miriam LEE,Young-Joon KO,Sangeun LEE,Daejin KIM,Hyungjun KIM,Sukmo KANG,Soyoung LEE,Jinjoo KIM,Sun Chang KIM,Youngsoo JUN,Sangyong JON 한국생물공학회 2015 한국생물공학회 학술대회 Vol.2015 No.4