Antigenic Differences Among Some Classical Tetrahymena pyriformis and T. vorax Strains*

SYNOPSIS. Suspensions of whole, killed ciliates were diffused against rabbit antisera for the respective strains to observe cross precipitation. The strains fell into the following groups: I. GL, H, ChS, GP, Aq, L-I, L-II, L-2, and V₁; II. W, L-3, Gl-R, and V₂; III. PR and F; and IV. BF and Lava. T. vorax strains V₁ and V₂ each resembled certain T. pyriformis strains more closely than they resembled each other. The same grouping of strains emerged in comparing antigenic suspensions and in comparing antisera and was confirmed by comparing absorption properties of antigens from the different strains.     

Electrophoretic Characterization of Classical Tetrahymena pyriformis Strains*

The electrophoretic mobility patterns of 8 enzymes have been examined in 43 classical strains of Tetrahymena pyriformis. The strains may be assorted into sets on the basis of a high degree of similarity of their mobility patterns. Strains of similar designation are frequently in different sets, whereas differently labeled strains may be in the same set. It is proposed that new strain designations be made on the basis of phenotypic similarity.     

Further Electrophoretic Characterization of Strains of Tetrahymena pyriformis*

Mobility patterns of 5 isozymes in strains of Tetrahymena pyriformis were demonstrated using polyacrylamide disc gel electrophoresis. Six stock strains were compared in these patterns to 4 strains representative of each of the previously described 4 major “phenotypic sets.” Stock strains segregated into predicted “phenosets,” and essentially confirmed validity and reproducibility of such a discrimination method. The proposal that new strain designations be assigned on a basis of “phenoset” conformity is reaffirmed.     

Biochemical and Ultrastructural Changes in Tetrahymena pyriformis During Starvation*

SYNOPSIS. Certain of the ultrastructural and biochemical changes occurring during the first 25 hr of starvation in Tetrahymena pyriformis were studied. Ultrastructurally, numerous profiles of degenerating mitochondria were seen in the early stages of starvation. The presence of oxidizable substrate such as glucose and acetate did not prevent this degeneration. Numerous large nucleoli were formed, many of which seemed to be passing into the cytoplasm as forming autophagic vacuoles. There was a transient increase in Oil Red O-positive bodies, presumably lipid (triglycerides). The extent and duration of this increase were pronounced in the presence of acetate. The lipid droplets appeared to arise within the cisternae of the endoplasmic reticulum. Lipid reserves were apparently utilized prior to carbohydrates, as the disappearance of lipid droplets preceded glycogen utilization, both in the presence of acetate and in the absence of exogenous substrate. A considerable loss of cellular protein also occurred. In cells from inorganic medium supplemented with glucose, glycogen occupied much of the cell, leaving only islands of cell organelles. Acid phosphatase was localized, ultrastructurally, mainly in autophagic vacuoles which contained mitochondria and other cell organelles, and in association with small, double-membraned structures which seemed to be sequestering small areas of cytoplasm. Such sequestered areas also appeared within larger autophagic vacuoles. Residual bodies containing concentric whorls of myelin-like membranes surrounding a more solid core accumulated during starvation. Acid phosphatase activity decreased in amount but not in specific activity. The specific activity of cathespin doubled or tripled, but there was little change in total enzyme.     

Cortical Characteristics of Strains of Syngens 10, 11 and 12 of Tetrahymena pyriformis*

SYNOPSIS. The patterns of correlation among cortical structures have been examined in strains of 3 additional breeding groups (syngens) of Tetrahymena pyriformis. As in previous studies, many cytogeometric properties were found to vary within a syngen. Surprisingly, one strain of syngen 12 differed from 3 others in the one property previously found to be constant—the pattern of variation of the position of the contractile vacuole pores. This strain may, however, be a sexually cross-reacting representative of still another syngen. This interpretation would preserve the diagnostic value of CVP position, and permit an ordering of syngens in terms of the distance of the CVP's from meridian #1: 1, 3, 7, 6, 8, 11, 12, 9, 2, 5, 4, 10.     

Macronuclear Cytology of Synchronized Tetrahymena pyriformis*

SYNOPSIS. Heat shock and stationary-phase conditions both cause fusion of nucleoli. In both cases the process is reversed when the cell is returned to normal physiological growth conditions. Fusion of nucleoli during the cell cycle of logarithmically growing cells was not observed. Likewise, fusion of nucleoli was not observed when the Padilla and Cameron(8) method of synchronization was used. The macronuclei of cells synchronized by the 1 cold-shock per cycle method(8) more closely resembled macronuclei of log-phase cells than did the macronuclei of cells synchronized by the Scherbaum and Zeuthen(12) heat-shock method.     

Toxicity of Antioxidants to Tetrahymena pyriformis*

SYNOPSIS. The cytotoxicity of 97 antioxidants of various structural types was determined for Tetrahymena pyriformis grown in a peptone medium. Cytotoxicity fo Tetrahymena, generally, was positively associated with acute oral tomicoiciity to rats but not with antioxidant pdency as measured by the Tetrahymena photodynamic assay for antioxidants. Some commonly used hindered-phenol, lipidsoluble anttioxidants, e.g., 2,6-di-tert.-butyl-P-cresol (BHT), had low toxicity (to Tetrahymena but poor antioxidant activity by the photodynamic test. Nmdihydroguahetic acid, with low toxicity to the rat and high Coxicity (to Tetrahymena, had very high activity in the photodynamic test.     

Some Factors Governing Respiration,Glucose Metabolism and Iodoacetate Sensitivity in Tetrahymena pyriformis*

SYNOPSIS. The physiologic response of Tetrahymena pyriformis W to glucose, as measured by the respiratory rate in a buffered suspension, was found to be altered by exposure of the culture to this carbohydrate in the peptone-based growth medium. Glucose-grown cells had an elevated respiratory rate in the presence of glucose in the Tris-buffered suspension, while cells not grown with the glucose supplement required sodium and potassium for the response. Orthophosphate elevated the rate of oxygen consumption independently of the glucose effect. The endogenous respiratory of glucose and non-glucose cultured cells were significantly diffe The 2 types of cells responded differently to the cations, sod and potassium, and were not identical in sensitivity to iodoacet Iodoacetate poisoning was found to be due to inhibition of “uncompetitive” type, potentiated by glucose, orthophosphate, the absence of potassium.     

Variation in Glyoxylate Bypass Inducibility among Strains of Tetrahymena pyriformis

SYNOPSIS. Seven strains of Tetrahymena pyriformis were assayed for log phase activity of the glyoxylate bypass enzymes isocitrate lyase and malate synthase. In strains 6I, 6II, 6III, and W, isocitrate lyase was induced; in HS, neither enzyme was induced by acetate. During growth in glucose- or acetate-containing media, strains 6III and GL had 2 periods of increased glyoxylate bypass and isocitrate dehydrogenase enzyme activities. Enzyme activities reached a maximum at the end of log phase, declined until the middle of stationary phase, and then increased again to a maximum near the end of stationary phase.     

Ionic Resistance in Strains of the Tetrahymena pyriformis Complex1

ABSTRACT. Strains of Tetrahymena thermophila were examined in an attempt to establish what role certain ions (Na⁺, K⁺, Li⁺, Ba⁺⁺, Ca⁺⁺, Mg⁺⁺, Mn⁺⁺, Al⁺⁺⁺, Fe⁺⁺⁺) play in influencing cell survival time in a culture medium. In short-term experiments (20–30 min), cell survival time in a 1% peptone medium is directly related to the valence of the ion employed. Long-term observations (lasting up to five days) in a 1% peptone medium containing lower ion concentrations revealed that the effects on cell-cycle time are not correlated with the valence state of the ion. Comparisons were made among the ionic resistances of strains of T. thermophila, of T. pyriformis sensu stricto, and of two subspecies of T. pigmentosa. Strains within a species are highly correlated in their patterns of ionic response, while marked differences between species occur. The most distinctive group of strains examined came from one of the subspecies (syngen 6) of T. pigmentosa.     

A Comparison of Cortical Proteins in Tetrahymena vorax Microstomes and Macrostomes1

Proteins of surface membranes and surface-related cytoskeletons in Tetrahymena vorax microstomes and macrostomes were compared by one-dimensional SDS polyacrylamide gel electrophoresis to see if protein differences could be detected that correlate with the transformation from one phenotype to the other. Some differences were observed. However, these alterations appear to result from the heat-shock procedure used to synchronize the microstome-to-macrostome transition. The apparent lack of transformationspecific changes in cortical proteins is discussed. Similarities and differences between cytoskeletal proteins of T. pyriformis GL-C and T. vorax are also noted and discussed.     

Survival of Tetrahymena pyriformis and Paramecium aurelia Following Freezing*

SYNOPSIS. Tetrahymena pyriformis strains E, A-136 31C and IMT II survived freezing in 10% dimethylsulfoxide when the temperature was lowered to freezing at 4.5 C/min. Survival was then obtained for at least 128 days by lowering the temperature rapidly to 95°C. Of the 3 strains, T. pyriformis IMT II was most resistant to the effects of freezing. Its volume averaged about half that of either of the other strains and may have contributed to the differences in survival. In addition to differences among strains, a medium relatively low in the concentration of nutrients, a culture nearing peak population, and a rate of cooling of 4.5 C/min, all gave best survival. Paramecium aurelia regained motility after being frozen in 6 to 7.5% dimethylsulfoxide for as long as 7 days at either –27 or –196 C, but cultures were obtained only after storage for 20 min at –27 C. A concentration of 6 to 7.5% dimethyl-sulfoxide, cooling at 4.5 C/min, and culture media containing Aerobacter aerogenes or composed of a commercially available composition were all required for survival of P. aurelia.     

Cytotoxicity of Synthetic Fuel Products on Tetrahymena pyriformis. I. Phenol*

SYNOPSIS. Phenol is the major organic constituent of coal-conversion scrub water and is therefore a potential environmental contaminant. After Tetrahymena pyriformis strain GL-C, syngen 1 was exposed to phenol, its behavior, cytology, respiration, and growth rates were examined. Concentrations ≥ 75 mg/liter alter cell motility, shape, and contractile vacuole activity. O₂ uptake was abruptly reduced within 3 min of exposure to phenol in concentrations as low as 10 mg/liter. Concomitantly there was an increase in the electron density of the mitochondrial matrix. Recovery to normal rates of O₂ consumption was paralleled by a return to normal matrix density. Alterations of mucocysts, pellicle, and glycogen were also observed. The length of lag phase growth curves increased generally in proportion to concentration of toxicant. Phenol, however, did not affect the rate of cell multiplication during the exponential growth phase. The potential use of this system to examine the effects of other possible organic pollutants is discussed.     

Some Amino Acid-Folic Acid Interrelationships in Tetrahymena pyriformis

DL-serine, DL-methionine or DL-serine + DL-methionine in excess inhibited the growth of Tetrahymena pyriformis H. Excess serine was most inhibitory at high concentration of folic acid, whereas the effect of excess methionine or methionine + serine was most pronounced at low levels of folic acid. Inhibition due to excess serine was relieved by raising the level of methionine or by adding pyrimethamine to lower the effective folic acid level, and was intensified by adding Dl.-ethionine or by raising the level of folic acid. Similarly, inhibition due to excess methionine was relieved by supplying more serine or adding DL-ethionine (which reduced the amount of available methionine) and was intensified by adding pyrimethamine. Inhibition by excess methionine + serine was reversed by increasing threonine, provided there was ample guanine present. Low levels of guanine or the presence of 8-azaguanine prevented this reversal. Comparisons are made with the work of others.     

Some Effects of Chloramphenicol and Ethidium Bromide on Tetrahymena pyriformis

SYNOPSIS. Very similar results are obtained by the addition of 300 μg/ml chloramphenicol or of 15 μg/ml ethidium bromide to cultures of Tetrahymena pyriformis strain ST. In such cultures the exponential growth rates and the yields are reduced. Unlike the untreated ciliates, which retain the pyriform shape throughout the exponential and stationary phases, those exposed to either of the drugs become more spherical. In the control organisms progressively less surface area/unit of cell volume is exposed to the environment during the growth cycle; in the drug-treated ciliates this pattern is reversed. Increased endogenous respiration, reduced specific activities of states 3 and 4, and reduction in the levels of cytochromes a, a₃, b , and c + c ₁ are found in mitochondria from ciliates exposed to chloramphenicol and ethidium bromide.