Fine structure of kappa in Paramecium aurelia.
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The fine structure of the ciliate Stentor has been studied by means of the electron microscope and the results have been correlated with observations made on the living organism by means of light microscopy; special reference has been made to structural features which may be responsible for contraction and extension in Stentor. Descriptions have been given of the structure of the macronucleus, the vacuolated cytoplasm, mitochondria and the pellicle; a detailed study has also been made of the adoral membranelles. About 250 membranelles encircle the peristomal cap and each is composed of 3 rows of cilia, with 20 to 25 cilia in each row; a fibrillar root system connected with the membranelles depends into the endoplasm for about 20 micro and each is essentially in the shape of a fan, the terminal ends of each root bifurcating to connect to neighbouring roots. The membranelles thus form a cohesive unit and this morphological arrangement may have a bearing on the motion and coordination of the whole system. Two structural features extending throughout the length of the animal have been identified per cortical stripe in the body wall of Stentor; first, km fibres lying just beneath the pellicle are composed of stacks of fibrillar sheets and are identical with the birefringent fibres observed in the living animal. The individual fibrils of the sheets are in turn connected to the kinetosomes of the body cilia; thus the km fibres are homologous to kinetodesmata. Secondly, M bands lie beneath the km fibres and form an interconnected system in contact with the surrounding vacuolated cytoplasm; the thickness of the M bands is greatest at the base of a contracted animal. The contractile and extensile properties of these organelles have been discussed in the light of experimental results and theoretical considerations.
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The ribbon-like macronucleus of Euplotes eurystomus pinches in half amitotically at each cell division. Several hours before the actual division two lightly staining duplication bands (reorganization bands) appear at the ends of the nucleus and approach each other slowly, finally meeting near the middle. Distal to the bands, that is, in regions through which the bands have already passed, the concentration of DNA (Feulgen) and "histone" (alkaline fast green) is greater than in the central zone. These facts suggest the hypothesis that DNA-histone synthesis takes place in a sequential fashion starting at the tips of the nucleus and proceeding to the middle. That this hypothesis is correct is shown by autoradiographic and photometric observations. Tritium-labelled thymidine is incorporated only in a limited region immediately distal to the bands. The average amount of Feulgen dye bound by the nucleus rises as the duplication bands approach each other, and is double the presynthesis value by the time the bands meet. A similar rise in the alkaline fast green dye is seen in duplicating nuclei, although no completely post-synthesis values were obtained in this study. The quantitative data are consistent with the assumption that the macronucleus contains a number of DNA-histone "units," presumably chromosomes, each of which duplicates once and only once.
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1. The amount of visible or long ultraviolet light (UV) required to photoreactivate Colpidium colpoda injured with known dosages of short UV (2654 A) was determined. 2. The effect of the short UV was tested by the delay in division of exposed animals compared to controls. Photoreactivation was tested by the effect of postillumination on the delay of division of treated colpidia compared to controls. 3. Colpidia were used in two physiological states: well fed and starved in balanced medium for 48 hours. The latter are much more sensitive to short UV although less susceptible of photoreactivation. 4. Photoreactivation occurred over the entire span from 3350 A to 4350 A for the well fed colpidia, from 3130 A to 5490 (green) for starved colpidia. 5. The photoreactivating effect of a single quantum of blue (4350 A) or long UV (3660 A) delivered per quantum of 2654 A used to injure colpidia was too slight to be considered significant. The effect of 10 quanta was usually more pronounced, but only after 100 quanta had been delivered was the photoreactivation nearly maximal for well fed colpidia. 6. The quantum requirement for maximal photoreactivation of the starved animals was greater at all wave lengths tried: 3660, 4050, 4350, and 5460 A being of the order of 800 incident quanta per incident quantum of 2654 A. 7. The transmission of UV(2654 A), blue, yellow, and red light by a suspension of colpidia was determined. 8. Large dosages of blue, violet, or long UV were slightly injurious to starved colpidia. In a few cases large dosages of 3660 A killed starved colpidia, especially after a non-lethal dose of short UV(2654 A). 9. Photoreactivation seems to be a balance between the slight injurious effect produced by the visible light or UV of long wave lengths and the injury produced by short wave length UV. 10. Possible reasons for the large number of quanta of photoreactivating light required per quantum of short UV are discussed.
1. A pigment can be extracted from Blepharisma undulans by heat treatment of a concentrated suspension of the deeply pigmented animals. 2. In the presence of this pigment, various colorless protozoans are sensitized to light and killed if exposed long enough. The protozoans show a differential sensitivity, some being much more sensitive than others. 3. Bleached colorless blepharismas are not sensitized to their own pigment even after prolonged exposure in the most concentrated solutions available. 4. Blepharisma is also less sensitive than any of the protozoans tested to such photodynamic dyes as rose bengal. 5. The pigment is not extracted from wet blepharismas by non-polar solvents, but is readily extracted into such polar organic solvents as the alcohols. 6. When the alcohol extract is dried, the amorphous residue is readily soluble in a variety of organic solvents, but not in the most non-polar. 7. The pigment is highly stable in alcohol extract and has been kept in the dark for years. 8. The pigment is bleached by light in a photooxidation. 9. Absorption maxima are found at wave lengths 5800, 5400, 4800, and 3300 A, the latter being the largest. Similar peaks are found in alcohol and water solutions, although the heights are not exactly the same. In both alcoholic and aqueous solutions pH had an effect on the absorption spectrum. Heat has little effect but illumination with intense visible light or exposure to ultraviolet light bleaches the pigment with decreases in the characteristic peaks. 10. Preliminary absorption column experiments indicate a single pigment in the alcohol extract. 11. Experiments on the migration of zoopurpurin in agar and gelatin gels indicate that it diffuses at about the same rate as eosin and is therefore probably not a large molecule.
1. The effect of the nutritional state of Didinium nasutum on its resistance to short ultraviolet (UV) radiation (2654 A) and its recovery from the injury following illumination with visible light (4350 A, blue) was studied. 2. The resistance of a didinium to UV is considerably increased by feeding it a paramecium 15 to 60 minutes before exposure to UV. If fed just before exposure to UV, the resistance is less than that of an unfed control. 3. Photoreversal is only slightly greater in didinia fed after irradiation with UV but before exposure to visible light as compared to those fed after exposure to visible light. 4. Irradiated paramecia are eaten by didinia, provided they have not started to cytolyze. Didinia fed on irradiated paramecia divide at about the same rate as controls or slightly faster. 5. The available stock of Didinium declines in vigor with lapse of time after excystment, as measured by the time required for division. The sensitivity of Didinium to UV did not change essentially during the 5 month period over which tests were made. 6. The theoretical implications of the results are considered.
Regeneration of Blepharisma undulans variety japonicus from which the hypostome has been removed is retarded by dosages of 3000 to 4600 ergs/mm.(2) at wavelength 2654A most strongly when the fragment is exposed soon after cutting. Dosages greater than 4600 ergs/mm.(2) prevent regeneration. Regeneration is also retarded strongly when the Blepharisma are cut soon after irradiation. Starvation retards regeneration and potentiates the effect of ultraviolet radiations. Division after regeneration of Blepharisma is also retarded by ultraviolet radiations about equally, regardless of when the Blepharisma are cut indicating a more lasting effect of the radiations upon the cells. Blepharisma cut after irradiation usually recover from the effects of the radiations sooner than uncut individuals given the same dosage. Retardation of division by ultraviolet radiation is subject to photoreversal by visible light, especially in a nitrogen atmosphere, provided the ultraviolet dose is not excessive. Visible light alone if prolonged, retards regeneration or may even kill the cut fragments of Blepharisma.
The same dosage of ultraviolet (UV) radiation retards division of several protozoans more effectively when the light is intermittent than when it is continuous, and especially at temperatures of 25-35 degrees C. At lower temperatures the difference between the effects of intermittent and continuous radiations is less marked. Somewhat similar results were obtained with the ciliates Paramecium caudatum, Blepharisma japonicum, and Colpidium colpoda, the disparity between intermittent and continuous light decreasing in the order given. The data are taken to indicate that thermochemical dark reactions succeed the absorption of UV radiations by the cells. In Blepharisma, besides initial delay in division, the cells stop dividing after one or two divisions, a "stasis" ensuing. Stasis is marked when the cells are irradiated at higher temperatures but is slight when they are irradiated at low temperatures, as if the temperature-sensitive reaction involved stasis (in all cases cultures are grown at 25 degrees C). The data are related to findings in the literature.
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