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opencc-by-4.0Dec 2023View details →

Fig. 3 in ATP accumulation in early resting cyst formation towards cryptobiosis in Colpoda cucullus

Fig. 3. Measurement of the relative amount of ATP per 10,000 cells in vegetative cells and cells at 12 h after the onset of encystment induction. The columns and attached bars represent the means and standard errors of 9 identical replicates, respectively. Double asterisks represent significant differences at p <0.01.

opencc-by-4.0Dec 2023View details →

Fig. 2 in ATP accumulation in early resting cyst formation towards cryptobiosis in Colpoda cucullus

Fig. 2. Relative gene expression of ATP synthase beta chain by real time PCR analysis. The columns and attached bars represent the means and standard errors of 4 identical replicates, respectively. Double asterisks represent significant differences at p <0.01.

opencc-by-4.0Dec 2023View details →

Fig. 1 in ATP accumulation in early resting cyst formation towards cryptobiosis in Colpoda cucullus

Fig. 1. Visualization of the mitochondrial membrane potential of Colpoda vegetative cells using a Mito PT assay kit; the results from cells at 0 h and cells at 1–24 h after the onset of encystment induction are shown. Differential interference microscopic observation (A) and fluorescence microscopic observation (B). Cells that contain mitochondria with polarized inner membranes show orange fluorescence, whereas those with depolarized mitochondria show green fluorescence. The bar represents 100 μm.

opencc-by-4.0Dec 2019View details →

Fig. 1 in Evidence of Stress Recovery in Free-Living Ciliate Colpoda cucullus: The Repair Capability of Resting Cysts to Damage Caused by Gamma Irradiation

Fig. 1. Excystment assay of Colpoda wet cysts (A) and dry cysts (B). 'Non-irradiated' indicates non-irradiated cysts; 'irradiated' indicates cysts irradiated at 4000 Gy, and 'irradiated-incubated' indicates cysts irradiated at 4000 Gy and incubated for 12 hours before the induction of excystment. Time indicates the number of hours after the induction of excystment. Columns and attached bars correspond to the means and standard errors, respectively, of six measurements. Asterisks and double asterisks represent significant differences at p <0.05 and p <0.01 (Mann-Whitney U test), respectively.

Fig. 2 in Tolerance of Colpoda cucullus Nag-1 Resting Cysts and Presumed Structure for Protection against UV Light

Fig. 2. Nomarski images (A-1−F-1) and their fluorescence photomicrographs (A-2−F-2) showing the formation of the auto-fluorescent cyst wall and NSPs in the encysting cells (wet cysts) after the onset of encystment induction. The cyst age (3 h to 5 days; A–F) is given at the upper right of each photograph. ec/en: ectocyst-endocyst complex, ec: an ectocyst layer, le: lepidosomes, ma: macronucleus, mu: mucus, NSP: nuclei-surrounding particle.

Fig. 2 in Gamma Radiation Tolerance and Protein Carbonylation Caused by Irradiation of Resting Cysts in the Free-living Ciliated Protist Colpoda cucullus

Fig. 2. Analysis of proteins (Left panel) and protein carbonylation by ECL (Right panel) from non-irradiated and 4000 Gy irradiated cells. The samples in the lanes were from non-irradiated cells (NonIR), 4000 Gy irradiated cells (IR), and cells incubated for 12 h after 4000 Gy irradiation (IR incubated). The protein bands and ECL signals were measured and are shown in parentheses for each lane relative to the Non-IR sample.

Fig. 4 in Gamma Radiation Tolerance and Protein Carbonylation Caused by Irradiation of Resting Cysts in the Free-living Ciliated Protist Colpoda cucullus

Fig. 4. Relative viability of Colpoda vegetative cells, wet cysts, and dry cysts after gamma radiation doses of 0 (non-irradiated), 500, 1000, 2000, 3000, and 4000 Gy. The column heights and attached bars are the means and standard errors, respectively, of six measurements at each dose. Double asterisks indicate a significant difference at p <0.01 (Mann-Whitney U test).

Fig. 3 in Gamma Radiation Tolerance and Protein Carbonylation Caused by Irradiation of Resting Cysts in the Free-living Ciliated Protist Colpoda cucullus

Fig. 3. Excystment of Colpoda dry cysts, after gamma irradiation at 0 (non-irradiated), 500, 1000, 2000, 3000, and 4000 Gy, as a function of time after induction of excystment. The points and bars mark the means and standard errors, respectively, of six measurements at each dose. The excystment mean ± SE at 3, 6, 24, and 96 h after the induction of excystment is shown in (a), (b), (c), and (d), respectively. The column heights and attached bars in (a) to (d) are the means and standard errors, respectively, of six measurements at each dose. Asterisks and double asterisks indicate a significant difference at p <0.05 and p <0.01, respectively (Mann-Whitney U test).

Fig. 1 in Tolerance of Colpoda cucullus Nag-1 Resting Cysts and Presumed Structure for Protection against UV Light

Fig. 1. Photomicrographs of yellowish auto-fluorescent structures of the mature cysts (wet cysts) of C. cucullus Nag-1 aged>2 weeks by UV excitation (A−D) and electron micrographs showing auto-fluorescent nuclei-surrounding particles (NSPs) (E, F) and cyst wall components (G). A, B − A Nomarski image (A) of a mature resting cyst and its fluorescence photomicrograph (B) showing autofluorescent structures. cw: cyst wall, ma: macronucleus. C−D − A Nomarski image (C) of a mature resting cyst and its fluorescence photomicrograph (D) showing autofluorescent layers of cyst wall. ec/en: an ectocyst-endocyst layer complex, en: endocyst layer. E−F − Electron micrographs showing NSPs. G − Electron micrograph showing cyst wall components. ec: ectocyst layer, en-1: first-synthesized layer of endocyst, en-2–6: second – sixth layers of endocyst, m: plasma membrane.

Fig. 1 in Gamma Radiation Tolerance and Protein Carbonylation Caused by Irradiation of Resting Cysts in the Free-living Ciliated Protist Colpoda cucullus

Fig. 1. Excystment of Colpoda wet cysts, after gamma irradiation at 0 (non-irradiated), 500, 1000, 2000, 3000, and 4000 Gy, as a function of time after the induction of excystment. The points and bars mark the means and standard errors, respectively, of six measurements at each dose. The excystment mean ± SE at 3, 6, 9, and 36 h after induction of excystment is shown in (a), (b), (c), and (d), respectively. The column heights and attached bars in (a) to (d) are the means and standard errors, respectively, of six measurements. Asterisks and double asterisks indicate a significant difference at p <0.05 and p <0.01, respectively (Mann-Whitney U test).

Fig. 4 in Tolerance of Colpoda cucullus Nag-1 Resting Cysts and Presumed Structure for Protection against UV Light

Fig. 4. Nomarski image (A-1) and its fluorescence photomicrograph (A-2) of a vacant cyst (cyst wall sample) obtained from 1-weekaged mature cysts of C. cucullus Nag-1, and its absorption spectrum (B) obtained by a spectrophotometer equipped with an integrating sphere.

opencc-by-4.0Oct 2016View details →

Fig. 1 in Record of two soil ciliates (Ciliophora: Colpodea: Colpoda) from Korea

Fig. 1. Colpoda cucullus from live (A, DG), protargolimpregnated (B, C, H, I) and silver nitrateimpregnated (J) specimens. A, B, D, E, H. right lateral views. C, I. left lateral views. C. arrow indicates excretory pore. E. arrow marks contractile vacuole, arrowhead indicates notches. F. extrusome at the cell margin (arrows). I. arrow marks excretory pore. J. "Cucullustype" silverline system, arrows indicate granules. CV, contractile vacuole. lP, left oral polykinetid. Ma, macronucleus. Mi, micronucleus. rP, right oral polykinetid. Scale bars: 50 μm.

opencc-by-4.0Oct 2016View details →

Fig. 2 in Record of two soil ciliates (Ciliophora: Colpodea: Colpoda) from Korea

Fig. 2. Colpoda inflata from live (A, DH), protargolimpregnated (B, C, I, J) and silver nitrateimpregnated (K) specimens. A, B, D, E, I. right lateral views. C, J. left lateral views. C. arrow indicates excretory pore. D. arrow indicates notches. E. arrow marks contractile vacuole. F. extrusome at the cell margin (arrows), arrowheads indicate food vacuoles. G. right oral polykinetid (arrow) and left oral polykinetid (arrowhead). J. arrow marks excretory pore. K. "Cucullustype" silverline system, arrows indicate granules. CV, contractile vacuole. lP, left oral polykinetid. Ma, macronucleus. Mi, micronucleus. rP, right oral polykinetid. Scale bars: 50 μm.

opencc-by-4.0Dec 2013View details →

Fig. 1 in Evaluation of Intracellular Ca Concentration by Fura 2 Ratiometry in Encystment-induced Colpoda cucullus

Fig. 1. The elevation in the intracellular Ca2+ concentration (F /F 340 380 ratio) in encystment-induced C. cucullus. F340 and F380 are the fluorescence intensities of fura 2 excited at 340-nm and 380-nm light, respectively. The cells loaded with fura 2-AM were suspended in 1 mM Tris-HCl buffer (pH 7.2) containing 0.1 mM CaCl2 (open circles), the same buffer without addition of CaCl2 (closed squares or closed circles), and the same buffer containing 10 µM EGTA (open squares) at high (50,000 cells/ml) ('H') or low (2,000 cells/ ml) ('L') cell density. Open triangles show the F340/F380 ratios of the cells suspended in 1 mM Tris-HCl buffer (pH 7.2) without addition of 0.1 mM CaCl2 at a low cell density (2,000 cells/ml) together with a high-density (48,000 particles/ml) PLP (26 µm in diameter, Sigma-Aldrich) ('L/PLP'). Points and attached bars correspond to the means of 5 identical measurements and standard errors.

Fig. 1 in Antifreeze Water-Rich Dormant Cysts of the Terrestrial Ciliate Colpoda cucullus Nag-1 at -65 ℃: Possible Involvement of Ultra-Antifreeze Polysaccharides

Fig. 1. Tolerance and antifreeze activity of wet resting cysts of C. cucullus Nag-1 in response to cooling (−65℃) and their osmolality. (A-1) Tolerance of encysting cells at various encystment stages. The abscissa indicates the cyst age of encysting cells. The ordinate indicates the excystment rates (%) of resting cysts (aged 1 day or more) or viability (%) of vegetative cells. Closed and open circles show the excystment rate or viability of the cells cooled at −65℃ for 24 h or more and those without cooling, respectively. The viability (%) of frozen vegetative cells is expressed as a percentage of the total number of tested cells (> 50 cells). The rate of excystment was expressed as a percentage of the total number of observed cells (50 cells). Points and attached bars correspond to the means of 6 measurements and the standard errors (SE), respectively. In each set of experiments (i.e., the cooling group and control group experiments), the same lot samples were used. (A-1, inset photograph) 1-week-old cysts cooled at −65℃ for 24 h, and thawed at room temperature. 'a': a living cyst, 'b': a cyst whose cell body shrank and detached from the cyst wall (killed cysts). (A-2) Inhibition of ice crystal growth in cell fluid obtained from C. cucullus Nag-1 resting cysts (2 or more weeks old). Upper and lower photomicrographs are pure water and cell fluid obtained from Colpoda cysts cooled at −65℃ for 30 min. (B-1) Changes in cell size of 2-week-old cysts after transfer from encystment-inducing medium without sucrose to medium containing 1 M sucrose. One run of measurement was done in the same cells. Points and attached bars correspond to the mean diameter obtained from 5 cells and the SE, respectively. (B-1, inset photograph) A 1-week-old resting cyst kept in encystment-inducing medium (left), and the same cyst transferred and kept for 5 min in the encystment-inducing medium containing 1 M sucrose (right). (B-2) Cell size of 2-week-old resting cysts immersed for 10 min in encystment-inducing medium containing various concentrations of sucrose. One run of measurement (0, 0.05, 0.1, 0.3, 1 M sucrose) was done in the same cell. Points and attached bars correspond to the mean diameter obtained from 10 cells and the SE, respectively.

Fig. 3 in Antifreeze Water-Rich Dormant Cysts of the Terrestrial Ciliate Colpoda cucullus Nag-1 at -65 ℃: Possible Involvement of Ultra-Antifreeze Polysaccharides

Fig. 3. PAS-stained SDS-PAGE gel (left) analyzing total proteins in encystment-induced cells, and CBB-staining of PAS-stained gel (right).

Fig. 8. A-D in Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of Colpoda cucullus Nag-1 and Cytoskeletal Dynamics

Fig. 8. A-D − Nomarski (leftmost lane), FITC-immunofluorescence images labeled with anti-α-tubulin monoclonal antibody and their magnified images (middle two lanes), and red fluorescence images (rightmost lane) stained with Acti-stain 555 phalloidin (detection for F-actin) of encysting cells of C. cucullus Nag-1. Each set of photomicrographs arranged in a horizontal row shows an identical cell except for Fig. 8C (FITC image, inset). A − Vegetative cell. B-D − Encysting cells of C. cucullus Nag-1 at 1.5 h (B), 3 h (C) and 3 days (D) after encystment induction. E − Nomarski image (left), red fluorescence images (middle) stained with Acti-stain 555 phalloidin, and a Nomarski image superimposed with a red fluorescence image obtained by Acti-stain 555 phalloidin staining (right) in encysting cells of C. cucullus Nag-1 at 3 h after encystment induction. F − Silver impregnation of a 3-day-aged cyst showing the basal structure of cilia. This photograph was reproduced from our previous work (Watoh et al. 2005, Fig. 9b). ant: anterior end, le: lepidosome, mu: mucus layer, ec/en: ectocyst layer lined with endocyst layer, m: plasma membrane. B − arrowheads: swollen tip of cilia. C − arrowhead: oral apparatus.

Fig. 6 in Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of Colpoda cucullus Nag-1 and Cytoskeletal Dynamics

Fig. 6. Ca2+/overpopulation-stimulated in vivo phosphorylation of p43 (actin, identified by MS) during resting cyst formation of C. cucullus Nag-1, detected by biotinylated Phos-tag/ECL assays (A), and blots stained with CBB after the biotinylated Phos-tag/ECL detection (B). Figures above the photographs indicate time lapse after onset of encystment induction.

Fig. 5 in Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of Colpoda cucullus Nag-1 and Cytoskeletal Dynamics

Fig. 5. Photomicrographs (Nomarski images) (A) and transmission electron micrographs (B) of C. cucullus Nag-1 after onset of encystment induction, showing resorption of cilia. (A) Vegetative cell at 0 h (A-1) and 2.5 h (A-2) after onset of encystment induction. (B) Encysting 3-h-aged cell (B-1) and 4-h-aged cell (B-2). ci: cilia, m: plasma membrane, ec: ectocyst layer, le: lepidosome. (B-2) a different electron micrograph of the same ultrathin section used in a previous paper (Funatani et al. 2010; Fig. 3).

Fig. 3 in Analysis of Water-Soluble Proteins by Two-Dimensional Electrophoresis in the Encystment Process of Colpoda cucullus Nag-1 and Cytoskeletal Dynamics

Fig. 3. Immunoblotting assay using anti α-tubulin antibody showing total α-tubulin content during resting cyst formation of C. cucullus Nag-1. Figures above the photographs indicate time lapse after on- set of encystment induction.