Subcellular Effects of Cytochalasin B and Dimethylsulfoxide on Paramecium aurelia*

SYNOPSIS. Paramecium aurelia syngen 4, stock 57 (sensitive) cultivated in Cerophyl infusion were exposed to cytochalasin B CB and dimethylsulfoxide (DMSO), the solvent for CB, to distinguish between the effects of these agents on a cellular system. DMSO significantly inhibited survival, fission rate, [³H]leucine incorporation, and cell size. CB-treated cells generally had slower division and poorer survival rates than cells exposed to the equivalent DMSO concentration, although the [3H]leucine incorporation was generally greater at the lower CB concentrations than for DMSO alone. As seen by electron microscopy and a new grycerination technic for observing polysomes, DMSO caused nuclear (nucleolar, chromatin) abnormalities as well as membrane degradation and polysomal breakdown; CB caused the formation of aberrant membrane structures and ribosomal tetramers, crystals, and tubes.     

Selective Autolysis of Nuclei as a Source of DNA Precursors in Paramecium aurelia Exconjugants*

The fate of labeled DNA in macronuclear fragments of starving Paramecium aurelia exconjugants was studied by quantitative autoradiography. Labeled material originally contained in DNA of macronuclear fragments is incorporated into macronuclear anlagen. During the starvation period the mean number of macronuclear fragments per cell decreased exponentially while there was an approximately exponential increase in the volume of macronuclear anlagen. Fragments appeared to be selectively and individually autolyzed. Labeled material originally contained in fragments was largely if not completely conserved through 108 hr of starvation during which more than 90% of the fragments were lost. Soluble labeled material was detectable after autolysis of fragments began, but finally almost all labeled material was incorporated into macronuclear anlagen.     

Effects of Actinomycin D on Generation Time and Morphogenesis in Paramecium*

SYNOPSIS. The sensitivity of Paramecium tetraurelia (=P. aurelia syngen 4) cells to pulse treatments with various doses of Actinomycin D (AMD) was estimated by comparing the generation times of treated and untreated sister cells. It was found that the delay of division in treated cells depended on the concentration of AMD, on their “age” at the time of the pulse treatment, and on their individual sensitivity. Sensitivity of Paramecium to AMD changes during the cell cycle in a predictable way. About 3 1/2 hr before the normally expected cell fission (total generation time ～ 5 1/2 hr) there is a decrease of sensitivity. Thereafter, the cell enters a new stage with a progressive increase of sensitivity. This 2nd phase ends at the “transition point” (～ 2 hr before cell division), when sensitivity drops abruptly. The division process itself may be altered and slowed down by high concentrations of AMD, even if the drug is applied after the transition point, but this process can never be completely annulled. The impairment of the division mechanism may lead to morphologic anomalies in the offspring. Resorption of oral anlagen in P. tetraurelia probably never occurs during the cell cycle after AMD treatment. The reason for individual variability of the cells, mechanisms controlling development, and the question of an obligate sequence of gene action in each cell cycle are discussed.     

The Synthesis of the Immobilization Antigen in Paramecium aurelia: In vitro Localization of Antigen in Ribosomal Cell Fractions

SYNOPSIS. The distribution of a cell surface protein, the immobilization antigen, has been studied in homogenates of Paramecium aurelia stock 168, syngen 1. Two fractions, a 9,000 g supernatant fluid and a 25,000 g supernatant fluid, have been investigated for their ability to bind ¹²⁵I-labelled antibodies specific against immobilization antigen. These 2 fractions have been characterized by electron microscopy. Antigen was detected in a pure microsome fraction and in the polysome fraction. Sucrose gradient analysis of the polysome fractions indicates that antigen is associated specifically with polysomes of 228S and 311S, the latter being located primarily in the microsome fraction. These results correlate well with the structure of the immobilization antigen proposed by Steers. A possible mechanism for the synthesis of this protein is outlined.     

Mutations Affecting the Trichocysts in Paramecium aurelia. I. Morphology and Description of the Mutants*

Six types of genic mutants have been isolated. Their phenotypes range from animals with no trichocysts (trichless), to animals with morphologically abnormal trichocysts (football, stubby, pointless, screwy-cigar), to animals which are incapable of extruding otherwise normal looking trichocysts (nondischarge). The football mutant possesses football-shaped trichocysts, which, unlike wild-type trichocysts, do not attach at the cortex. The stubby mutant possesses shorter trichocysts which have a very highly variable morphology. The screwy-cigar animals have thinner and usually longer trichocysts than those found in wild-type cells. The trichocysts of the pointless mutant have all the components of the wild-type organelles but not in their proper relationship. Electron microscopic studies of the mutants have demonstrated that although the morphology of the various mutant trichocysts may differ, their ultrastructure and early developmental stages are comparable to those of trichocysts found in wild type. The mutations are usually pleiotropic, affecting other systems besides trichocysts. The existence of these mutants, particularly trichless, poses some interesting questions regarding the function of trichocysts, and also gives insight into the development of trichocysts.