Yoichiro Sogame, K. Kojima, T. Takeshita, Shiho Kikuchi, Yuto Shimada, Rikiya Nakamura, M. Arikawa, S. Miyata, E. Kinoshita, F. Suizu, T. Matsuoka
{"title":"包囊过程中水溶性蛋白的双向电泳分析及细胞骨架动力学","authors":"Yoichiro Sogame, K. Kojima, T. Takeshita, Shiho Kikuchi, Yuto Shimada, Rikiya Nakamura, M. Arikawa, S. Miyata, E. Kinoshita, F. Suizu, T. Matsuoka","doi":"10.4467/16890027AP.20.009.13264","DOIUrl":null,"url":null,"abstract":"Assays of protein contained in water-soluble fraction of encysting cells Colpoda cucullus Nag-1 by two-dimensional electrophoresis (2-D PAGE) and mass spectrometry (MS) revealed that the amount of β-tubulin abruptly increased in 2.5–10 h after encystment induction. Judging from the results that total α-tubulin content did not decrease much until 12 h after encystment induction, the result indicates that disassembly of microtubules may occur soon after encystment is induced. Therefore, we tried to visualize dynamics of microtubules. Immunofluorescence microscopy using anti-α-tubulin antibody indicated that disassembly of axonemal microtubules of cilia became within 1.5 h after encystment induction, and resorbed in 3 days. Although the cytoplasmic microtubules failed to be visualized clearly, encystmentdependent globulation of cells was promoted by taxol, an inhibitor of disassembly of microtubules. It is possible that a temporary formation of cytoplasmic microtubules may be involved in cell globulation.\n\nThe phosphorylation level of actin (43 kDa) became slightly elevated just after encystment induction. Lepidosomes, the sticky small globes surrounding encysting cells, were vividly stained with Acti-stain 555 phalloidin, suggesting that 43-kDa actin or its homologues may be contained in lepidosomes.","PeriodicalId":50883,"journal":{"name":"Acta Protozoologica","volume":"1 1","pages":""},"PeriodicalIF":1.5000,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"2","resultStr":"{\"title\":\"Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of Colpoda cucullus Nag-1 and Cytoskeletal Dynamics\",\"authors\":\"Yoichiro Sogame, K. Kojima, T. Takeshita, Shiho Kikuchi, Yuto Shimada, Rikiya Nakamura, M. Arikawa, S. Miyata, E. Kinoshita, F. Suizu, T. Matsuoka\",\"doi\":\"10.4467/16890027AP.20.009.13264\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Assays of protein contained in water-soluble fraction of encysting cells Colpoda cucullus Nag-1 by two-dimensional electrophoresis (2-D PAGE) and mass spectrometry (MS) revealed that the amount of β-tubulin abruptly increased in 2.5–10 h after encystment induction. Judging from the results that total α-tubulin content did not decrease much until 12 h after encystment induction, the result indicates that disassembly of microtubules may occur soon after encystment is induced. Therefore, we tried to visualize dynamics of microtubules. Immunofluorescence microscopy using anti-α-tubulin antibody indicated that disassembly of axonemal microtubules of cilia became within 1.5 h after encystment induction, and resorbed in 3 days. Although the cytoplasmic microtubules failed to be visualized clearly, encystmentdependent globulation of cells was promoted by taxol, an inhibitor of disassembly of microtubules. It is possible that a temporary formation of cytoplasmic microtubules may be involved in cell globulation.\\n\\nThe phosphorylation level of actin (43 kDa) became slightly elevated just after encystment induction. Lepidosomes, the sticky small globes surrounding encysting cells, were vividly stained with Acti-stain 555 phalloidin, suggesting that 43-kDa actin or its homologues may be contained in lepidosomes.\",\"PeriodicalId\":50883,\"journal\":{\"name\":\"Acta Protozoologica\",\"volume\":\"1 1\",\"pages\":\"\"},\"PeriodicalIF\":1.5000,\"publicationDate\":\"2021-03-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"2\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Acta Protozoologica\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.4467/16890027AP.20.009.13264\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Acta Protozoologica","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.4467/16890027AP.20.009.13264","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of Colpoda cucullus Nag-1 and Cytoskeletal Dynamics
Assays of protein contained in water-soluble fraction of encysting cells Colpoda cucullus Nag-1 by two-dimensional electrophoresis (2-D PAGE) and mass spectrometry (MS) revealed that the amount of β-tubulin abruptly increased in 2.5–10 h after encystment induction. Judging from the results that total α-tubulin content did not decrease much until 12 h after encystment induction, the result indicates that disassembly of microtubules may occur soon after encystment is induced. Therefore, we tried to visualize dynamics of microtubules. Immunofluorescence microscopy using anti-α-tubulin antibody indicated that disassembly of axonemal microtubules of cilia became within 1.5 h after encystment induction, and resorbed in 3 days. Although the cytoplasmic microtubules failed to be visualized clearly, encystmentdependent globulation of cells was promoted by taxol, an inhibitor of disassembly of microtubules. It is possible that a temporary formation of cytoplasmic microtubules may be involved in cell globulation.
The phosphorylation level of actin (43 kDa) became slightly elevated just after encystment induction. Lepidosomes, the sticky small globes surrounding encysting cells, were vividly stained with Acti-stain 555 phalloidin, suggesting that 43-kDa actin or its homologues may be contained in lepidosomes.
期刊介绍:
Acta Protozoologica - International Journal on Protistology - is a quarterly journal that publishes current and comprehensive, experimental, and theoretical contributions across the breadth of protistology, and cell biology of Eukaryote microorganisms including: behaviour, biochemistry and molecular biology, development, ecology, genetics, parasitology, physiology, photobiology, systematics and phylogeny, and ultrastructure. It publishes original research reports, critical reviews of current research written by invited experts in the field, short communications, book reviews, and letters to the Editor.