Cell structure and function最新文献_第10页

An Anti-tumorigenic Role of the Warburg Effect at Emergence of Transformed Cells. Warburg效应在转化细胞出现中的抗肿瘤作用。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2018-09-01 Epub Date: 2018-07-26 DOI: 10.1247/csf.18018

Kojiro Ishibashi, Riku Egami, Kazuki Nakai, Shunsuke Kon

引用次数: 6

Automatic Quantitative Segmentation of Myotubes Reveals Single-cell Dynamics of S6 Kinase Activation. 肌管自动定量分割揭示S6激酶激活的单细胞动力学。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2018-08-31 Epub Date: 2018-07-26 DOI: 10.1247/csf.18012

Haruki Inoue, Katsuyuki Kunida, Naoki Matsuda, Daisuke Hoshino, Takumi Wada, Hiromi Imamura, Hiroyuki Noji, Shinya Kuroda

{"title":"Automatic Quantitative Segmentation of Myotubes Reveals Single-cell Dynamics of S6 Kinase Activation.","authors":"Haruki Inoue, Katsuyuki Kunida, Naoki Matsuda, Daisuke Hoshino, Takumi Wada, Hiromi Imamura, Hiroyuki Noji, Shinya Kuroda","doi":"10.1247/csf.18012","DOIUrl":"https://doi.org/10.1247/csf.18012","url":null,"abstract":"Automatic cell segmentation is a powerful method for quantifying signaling dynamics at single-cell resolution in live cell fluorescence imaging. Segmentation methods for mononuclear and round shape cells have been developed extensively. However, a segmentation method for elongated polynuclear cells, such as differentiated C2C12 myotubes, has yet to be developed. In addition, myotubes are surrounded by undifferentiated reserve cells, making it difficult to identify background regions and subsequent quantification. Here we developed an automatic quantitative segmentation method for myotubes using watershed segmentation of summed binary images and a two-component Gaussian mixture model. We used time-lapse fluorescence images of differentiated C2C12 cells stably expressing Eevee-S6K, a fluorescence resonance energy transfer (FRET) biosensor of S6 kinase (S6K). Summation of binary images enhanced the contrast between myotubes and reserve cells, permitting detection of a myotube and a myotube center. Using a myotube center instead of a nucleus, individual myotubes could be detected automatically by watershed segmentation. In addition, a background correction using the two-component Gaussian mixture model permitted automatic signal intensity quantification in individual myotubes. Thus, we provide an automatic quantitative segmentation method by combining automatic myotube detection and background correction. Furthermore, this method allowed us to quantify S6K activity in individual myotubes, demonstrating that some of the temporal properties of S6K activity such as peak time and half-life of adaptation show different dose-dependent changes of insulin between cell population and individuals.Key words: time lapse images, cell segmentation, fluorescence resonance energy transfer, C2C12, myotube.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"43 2","pages":"153-169"},"PeriodicalIF":1.5,"publicationDate":"2018-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1247/csf.18012","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36342690","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Branched Sialylated N-glycans Are Accumulated in Brain Synaptosomes and Interact with Siglec-H. 分支唾液化n -聚糖在脑突触体中积累并与siglece - h相互作用。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2018-08-25 Epub Date: 2018-07-20 DOI: 10.1247/csf.18009

Mai Handa-Narumi, Takeshi Yoshimura, Hiroyuki Konishi, Yuko Fukata, Yoshiyuki Manabe, Katsunori Tanaka, Guang-Ming Bao, Hiroshi Kiyama, Koichi Fukase, Kazuhiro Ikenaka

{"title":"Branched Sialylated N-glycans Are Accumulated in Brain Synaptosomes and Interact with Siglec-H.","authors":"Mai Handa-Narumi, Takeshi Yoshimura, Hiroyuki Konishi, Yuko Fukata, Yoshiyuki Manabe, Katsunori Tanaka, Guang-Ming Bao, Hiroshi Kiyama, Koichi Fukase, Kazuhiro Ikenaka","doi":"10.1247/csf.18009","DOIUrl":"https://doi.org/10.1247/csf.18009","url":null,"abstract":"Proper N-glycosylation of proteins is important for normal brain development and nervous system function. Identification of the localization, carrier proteins and interacting partners of N-glycans is essential for understanding the roles of glycoproteins. The present study examined the N-glycan A2G'2F (Galβ1-3GlcNAcβ1-2Manα1-6[Galβ1-3GlcNAcβ1-2Manα1-3]Manβ1-4GlcNAcβ1-4[Fucα1-6]GlcNAc-). A2G'2F has a branched sialic acid structural feature, and branched sialylated A2G'2F is a major N-glycan in the mouse brain. Its expression in the mouse brain increases during development, suggesting that branched sialylated N-glycans play essential roles during brain development. However, the carrier proteins, interacting partners and localization of branched sialylated N-glycans remain unknown. We previously improved our method for analyzing N-glycans from trace samples, and here we succeeded in detecting A2G'2F in small fragments excised from the two-dimensional electrophoresis gels of subcellular fractionated mouse brain proteins. A2G'2F was accumulated in mouse brain synaptosomes. We identified calreticulin as one of the candidate A2G'2F carriers and found calreticulin expression in both the endoplasmic reticulum and synaptosomal fractions. Calreticulin was observed in dendritic spines of cultured cortical neurons. Synthesized branched sialylated glycan clusters interacted with sialic acid-binding immunoglobulin-like lectin H (Siglec-H), which is known to be a microglia-specific molecule. Taken together, these results suggest that branched sialylated A2G'2F in synaptosomes plays a role in the interaction of dendritic spines with microglia.Key words: N-glycan, subcellular fractionation, calreticulin, dendritic spine, Siglec-H.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"43 2","pages":"141-152"},"PeriodicalIF":1.5,"publicationDate":"2018-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1247/csf.18009","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36335301","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10

A Novel Morphological Marker for the Analysis of Molecular Activities at the Single-cell Level. 在单细胞水平上分析分子活动的一种新的形态标记。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2018-08-10 Epub Date: 2018-06-29 DOI: 10.1247/csf.18013

Ayako Imanishi, Tomokazu Murata, Masaya Sato, Kazuhiro Hotta, Itaru Imayoshi, Michiyuki Matsuda, Kenta Terai

{"title":"A Novel Morphological Marker for the Analysis of Molecular Activities at the Single-cell Level.","authors":"Ayako Imanishi, Tomokazu Murata, Masaya Sato, Kazuhiro Hotta, Itaru Imayoshi, Michiyuki Matsuda, Kenta Terai","doi":"10.1247/csf.18013","DOIUrl":"https://doi.org/10.1247/csf.18013","url":null,"abstract":"For more than a century, hematoxylin and eosin (H&E) staining has been the de facto standard for histological studies. Consequently, the legacy of histological knowledge is largely based on H&E staining. Due to the recent advent of multi-photon excitation microscopy, the observation of live tissue is increasingly being used in many research fields. Adoption of this technique has been further accelerated by the development of genetically encoded biosensors for ions and signaling molecules. However, H&E-based histology has not yet begun to fully utilize in vivo imaging due to the lack of proper morphological markers. Here, we report a genetically encoded fluorescent marker, NuCyM (Nucleus, Cytosol, and Membrane), which is designed to recapitulate H&E staining patterns in vivo. We generated a transgenic mouse line ubiquitously expressing NuCyM by using a ROSA26 bacterial artificial chromosome (BAC) clone. NuCyM evenly marked the plasma membrane, cytoplasm and nucleus in most tissues, yielding H&E staining-like images. In the NuCyM-expressing cells, cell division of a single cell was clearly observed as five basic phases during M phase by three-dimensional imaging. We next crossed NuCyM mice with transgenic mice expressing an ERK biosensor based on the principle of Förster resonance energy transfer (FRET). Using NuCyM, ERK activity in each cell could be extracted from the FRET images. To further accelerate the image analysis, we employed machine learning-based segmentation methods, and thereby automatically quantitated ERK activity in each cell. In conclusion, NuCyM is a versatile cell morphological marker that enables us to grasp histological information as with H&E staining.Key words: in vivo imaging, histology, machine learning, molecular activity.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"43 2","pages":"129-140"},"PeriodicalIF":1.5,"publicationDate":"2018-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1247/csf.18013","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36274728","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 15

Detailed Analysis of the Interaction of Yeast COG Complex. 酵母COG复合物相互作用的详细分析。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2018-07-19 Epub Date: 2018-06-14 DOI: 10.1247/csf.18014

Midori Ishii, Vladimir V Lupashin, Akihiko Nakano

{"title":"Detailed Analysis of the Interaction of Yeast COG Complex.","authors":"Midori Ishii, Vladimir V Lupashin, Akihiko Nakano","doi":"10.1247/csf.18014","DOIUrl":"https://doi.org/10.1247/csf.18014","url":null,"abstract":"The Golgi apparatus is a central station for protein trafficking in eukaryotic cells. A widely accepted model of protein transport within the Golgi apparatus is cisternal maturation. Each cisterna has specific resident proteins, which are thought to be maintained by COPI-mediated transport. However, the mechanisms underlying specific sorting of these Golgi-resident proteins remain elusive. To obtain a clue to understand the selective sorting of vesicles between the Golgi cisterenae, we investigated the molecular arrangements of the conserved oligomeric Golgi (COG) subunits in yeast cells. Mutations in COG subunits cause defects in Golgi trafficking and glycosylation of proteins and are causative of Congenital Disorders of Glycosylation (CDG) in humans. Interactions among COG subunits in cytosolic and membrane fractions were investigated by co-immunoprecipitation. Cytosolic COG subunits existed as octamers, whereas membrane-associated COG subunits formed a variety of subcomplexes. Relocation of individual COG subunits to mitochondria resulted in recruitment of only a limited number of other COG subunits to mitochondria. These results indicate that COG proteins function in the forms of a variety of subcomplexes and suggest that the COG complex does not comprise stable tethering without other interactors.Key words: The Golgi apparatus, COG complex, yeast, membrane trafficking, multi-subunit tethering complex.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"43 2","pages":"119-127"},"PeriodicalIF":1.5,"publicationDate":"2018-07-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1247/csf.18014","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36219302","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 6

Solo and Keratin Filaments Regulate Epithelial Tubule Morphology. 单丝和角蛋白丝调节上皮小管形态。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2018-06-02 Epub Date: 2018-04-28 DOI: 10.1247/csf.18010

Ryosuke Nishimura, Kagayaki Kato, Sachiko Fujiwara, Kazumasa Ohashi, Kensaku Mizuno

{"title":"Solo and Keratin Filaments Regulate Epithelial Tubule Morphology.","authors":"Ryosuke Nishimura, Kagayaki Kato, Sachiko Fujiwara, Kazumasa Ohashi, Kensaku Mizuno","doi":"10.1247/csf.18010","DOIUrl":"https://doi.org/10.1247/csf.18010","url":null,"abstract":"Epithelial tubules, consisting of the epithelial cell sheet with a central lumen, are the basic structure of many organs. Mechanical forces play an important role in epithelial tubulogenesis; however, little is known about the mechanisms controlling the mechanical forces during epithelial tubule morphogenesis. Solo (also known as ARHGEF40) is a RhoA-targeting guanine-nucleotide exchange factor that is involved in mechanical force-induced RhoA activation and stress fiber formation. Solo binds to keratin-8/keratin-18 (K8/K18) filaments, and this interaction plays a crucial role in mechanotransduction. In this study, we examined the roles of Solo and K8/K18 filaments in epithelial tubulogenesis using MDCK cells cultured in 3D collagen gels. Knockdown of either Solo or K18 resulted in rounder tubules with increased lumen size, indicating that Solo and K8/K18 filaments play critical roles in forming the elongated morphology of epithelial tubules. Moreover, knockdown of Solo or K18 decreased the level of diphosphorylated myosin light chain (a marker of contractile force) at the luminal and outer surfaces of tubules, suggesting that Solo and K8/K18 filaments are involved in the generation of the myosin II-mediated contractile force during epithelial tubule morphogenesis. In addition, K18 filaments were normally oriented along the long axis of the tubule, but knockdown of Solo perturbed their orientation. These results suggest that Solo plays crucial roles in forming the elongated morphology of epithelial tubules and in regulating myosin II activity and K18 filament organization during epithelial tubule formation.Key words: epithelial tubulogenesis, Solo, keratin, Rho-GEF, myosin.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"43 1","pages":"95-105"},"PeriodicalIF":1.5,"publicationDate":"2018-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1247/csf.18010","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36056452","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 6

Hidden Peptides Encoded by Putative Noncoding RNAs. 由假定的非编码rna编码的隐藏肽。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2018-05-18 Epub Date: 2018-04-11 DOI: 10.1247/csf.18005

Akinobu Matsumoto, Keiichi I Nakayama

引用次数: 46

Live-cell Imaging with Genetically Encoded Protein Kinase Activity Reporters. 基因编码蛋白激酶活性报告基因的活细胞成像。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2018-04-25 Epub Date: 2018-03-17 DOI: 10.1247/csf.18003

Gembu Maryu, Haruko Miura, Youichi Uda, Akira T Komatsubara, Michiyuki Matsuda, Kazuhiro Aoki

{"title":"Live-cell Imaging with Genetically Encoded Protein Kinase Activity Reporters.","authors":"Gembu Maryu, Haruko Miura, Youichi Uda, Akira T Komatsubara, Michiyuki Matsuda, Kazuhiro Aoki","doi":"10.1247/csf.18003","DOIUrl":"https://doi.org/10.1247/csf.18003","url":null,"abstract":"Protein kinases play pivotal roles in intracellular signal transduction, and dysregulation of kinases leads to pathological results such as malignant tumors. Kinase activity has hitherto been measured by biochemical methods such as in vitro phosphorylation assay and western blotting. However, these methods are less useful to explore spatial and temporal changes in kinase activity and its cell-to-cell variation. Recent advances in fluorescent proteins and live-cell imaging techniques enable us to visualize kinase activity in living cells with high spatial and temporal resolutions. Several genetically encoded kinase activity reporters, which are based on the modes of action of kinase activation and phosphorylation, are currently available. These reporters are classified into single-fluorophore kinase activity reporters and Förster (or fluorescence) resonance energy transfer (FRET)-based kinase activity reporters. Here, we introduce the principles of genetically encoded kinase activity reporters, and discuss the advantages and disadvantages of these reporters.Key words: kinase, FRET, phosphorylation, KTR.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"43 1","pages":"61-74"},"PeriodicalIF":1.5,"publicationDate":"2018-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1247/csf.18003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35925190","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 21

SPIRAL2 Stabilises Endoplasmic Microtubule Minus Ends in the Moss Physcomitrella patens. SPIRAL2稳定苔藓小壶菌内质微管负端。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2018-03-28 Epub Date: 2018-03-15 DOI: 10.1247/csf.18001

Shu Yao Leong, Moé Yamada, Naoki Yanagisawa, Gohta Goshima

{"title":"SPIRAL2 Stabilises Endoplasmic Microtubule Minus Ends in the Moss Physcomitrella patens.","authors":"Shu Yao Leong, Moé Yamada, Naoki Yanagisawa, Gohta Goshima","doi":"10.1247/csf.18001","DOIUrl":"https://doi.org/10.1247/csf.18001","url":null,"abstract":"Stabilisation of minus ends of microtubules (MTs) is critical for organising MT networks in land plant cells, in which all MTs are nucleated independent of centrosomes. Recently, Arabidopsis SPIRAL2 (SPR2) protein was shown to localise to plus and minus ends of cortical MTs, and increase stability of both ends. Here, we report molecular and functional characterisation of SPR2 of the basal land plant, the moss Physcomitrella patens. In protonemal cells of P. patens, where non-cortical, endoplasmic MT network is organised, we observed SPR2 at minus ends, but not plus ends, of endoplasmic MTs and likely also of phragmoplast MTs. Minus end decoration was reconstituted in vitro using purified SPR2, suggesting that moss SPR2 is a minus end-specific binding protein (-TIP). We generated a loss-of-function mutant of SPR2, in which frameshift-causing deletions/insertions were introduced into all four paralogous SPR2 genes by means of CRISPR/Cas9. Protonemal cells of the mutant showed instability of endoplasmic MT minus ends. These results indicate that moss SPR2 is a MT minus end stabilising factor.Key words: acentrosomal microtubule network, microtubule minus end, P. patens, CAMSAP/Nezha/Patronin.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"43 1","pages":"53-60"},"PeriodicalIF":1.5,"publicationDate":"2018-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1247/csf.18001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35831847","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 27

A Missense Mutation in the NSF Gene Causes Abnormal Golgi Morphology in Arabidopsis thaliana. 拟南芥NSF基因错义突变导致高尔基体形态异常。

IF 1.5 4区生物学

Cell structure and function Pub Date : 2018-03-27 Epub Date: 2018-03-02 DOI: 10.1247/csf.17026

Sayuri Tanabashi, Keiko Shoda, Chieko Saito, Tomoaki Sakamoto, Tetsuya Kurata, Tomohiro Uemura, Akihiko Nakano

{"title":"A Missense Mutation in the NSF Gene Causes Abnormal Golgi Morphology in Arabidopsis thaliana.","authors":"Sayuri Tanabashi, Keiko Shoda, Chieko Saito, Tomoaki Sakamoto, Tetsuya Kurata, Tomohiro Uemura, Akihiko Nakano","doi":"10.1247/csf.17026","DOIUrl":"https://doi.org/10.1247/csf.17026","url":null,"abstract":"The Golgi apparatus is a key station of glycosylation and membrane traffic. It consists of stacked cisternae in most eukaryotes. However, the mechanisms how the Golgi stacks are formed and maintained are still obscure. The model plant Arabidopsis thaliana provides a nice system to observe Golgi structures by light microscopy, because the Golgi in A. thaliana is in the form of mini-stacks that are distributed throughout the cytoplasm. To obtain a clue to understand the molecular basis of Golgi morphology, we took a forward-genetic approach to isolate A. thaliana mutants that show abnormal structures of the Golgi under a confocal microscope. In the present report, we describe characterization of one of such mutants, named #46-3. The #46-3 mutant showed pleiotropic Golgi phenotypes. The Golgi size was in majority smaller than the wild type, but varied from very small ones, sometimes without clear association of cis and trans cisternae, to abnormally large ones under a confocal microscope. At the ultrastructual level by electron microscopy, queer-shaped large Golgi stacks were occasionally observed. By positional mapping, genome sequencing, and complementation and allelism tests, we linked the mutant phenotype to the missense mutation D374N in the NSF gene, encoding the N-ethylmaleimide-sensitive factor (NSF), a key component of membrane fusion. This residue is near the ATP-binding site of NSF, which is very well conserved in eukaryotes, suggesting that the biochemical function of NSF is important for maintaining the normal morphology of the Golgi.Key words: Golgi morphology, N-ethylmaleimide-sensitive factor (NSF), Arabidopsis thaliana.","PeriodicalId":9927,"journal":{"name":"Cell structure and function","volume":"43 1","pages":"41-51"},"PeriodicalIF":1.5,"publicationDate":"2018-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1247/csf.17026","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35793095","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 6