The fine structure of the pellicle ofEuglena gracilis as revealed by freeze-etching

Summary The ultrastructure of the pellicle ofEuglena gracilis (Klebs) Z strain was studied using the freeze-etching technique and the results were correlated with data obtained from thin sections of fixed material.Examination of freeze-etched pellicles reveals an outer particulate layer and an inner striated layer. The particles of the outer layer measure approximately 150 Å in diameter. The striations of the inner layer are about 50 Å wide and are separated from each other by about 35 Å. A broad repeating pattern is also visible with a periodicity of about 450 Å. When deep etching is employed, a smooth outer layer is seen covering the particulate layer. This is probably the outer surface of the plasma membrane. Mucilage is present on the outer surface of the cell and is seen as a substructure of threads superposed on the smooth layer of plasma membrane.Thin sectioning also shows a striated layer interior to the plasma membrane. This appears to be identical to the striated layer seen after freeze-etching.     

The Distribution of Tetrahymena pyriformis1

SYNOPSIS. This paper is a brief account of both amicronucleate and sexually active strains of Tetrahymena pyriformis and their distribution with some comments on their possible evolution.     

Cytoplasmic membrane changes in conjugating Tetrahymena

Summary The formation of stacks of smooth-surfaced cisternae was studied in samples of Tetrahymena pyriformis that were prepared for electron microscopy after starvation for 2 days in buffer, and in samples of organisms of different mating type that were starved for 2 days and then mixed to induce conjugation. The number of stacks of cisternae was greater in starved ciliates than in those from stock cultures, and the size and number of stacks increased further after mixing animals of different mating type. When cells were either starved or mixed in buffer to which the protein synthesis inhibitor cycloheximide had been added, the formation of the membranous stacks was almost completely abolished. Addition of the RNA synthesis inhibitor actinomycin D, however, did not result in a significant decrease in the size and number of the stacks of saccules. Conjugation did not occur in the presence of either cycloheximide or actinomycin. The results suggest that protein synthesis is required for the formation of the stacks, but RNA synthesis is no longer necessary for their formation at this stage. The previous identification of the stacks as a Golgi apparatus and the possible functions of these membranes are considered.Supported by grants from NSF (GB-32285) and the American Cancer Society (E-500).The authors acknowledge the technical assistance of Mrs. Sue Thompson.     

The transsulfuration pathway in Tetrahymena pyriformis.

Four enzymes necessary for the metabolism of methionine by the trans-sulfuration pathway, methionine adenosyltransferase (EC 2.5.1.6), adenosylhomocysteinase (EC 3.3.1.1), cystathionine beta-synthase (EC 4.2.1.22) and cystathionine gamma-lyase (EC 4.4.1.1) were identified in Tetrahymean pyriformis. The ability of these cells to transfer 35S from E135S]methionine to form [35S] cysteine was also observed and taken as direct evidence for the functional existence of this pathway in Tetrahymena. An intermediate in the pathway and an active methyl donor, S-adenosylmethionine, was qualitatively identified in Tetrahymena and its concentration was found to be greater in late stationary phase cells than in early stationary phase cells.     

Observations on the fine structure of the pellicle pores ofEuglena granulata

Summary The structure or granules associated with the pellicle strips ofEuglena granulata (Klebs) Schmitz has been studied using light and electron microscopy. Two types of bodies can be distinguished in this association on the basis of their staining reactions, distribution, ultrastructure, and other characteristics. The first type, called pellicle pores, is distributed with regularity along certain pellicle strips; the pores are very uniform in size (about 300 m in diameter) and in content. They consist of an aperture and a compartment, the latter lined by the plasmalemma; in each compartment a dense, osmiophilic body and a series of tubules occur. The structure of the pellicle strips is modified at the points where the pellicle pores are present; the aperture extends to the cell surface in the groove between two pellicle strips. The second type of body, which undergoes vital staining with neutral red, is called a pellicle vacuole. These are variable in size (ca. 1 to 2 microns in diameter) and contain a variety of components including myelin figures, proliferated membranes, and sheets of electrondense material. The structure of these elements as well as their possible functional significance are discussed in some detail.     

The Induction of Endocytosis in Starved Tetrahymena pyriformis

SYNOPSIS. The formation of digestive vacuoles by starved Tetrahymena pyriformis could be induced by mixtures of latex particles and a variety of potentially digestible solutes. Latex particles themselves had little effect in inducing vacuole formation. Protein, polypeptide, and RNA were highly effective inducers, while glutamate, amino acid mixtures, polysacharides, and glucose were moderately effective. Sodium-β-glycerophosphate had a slight effect and sodium acetate was ineffective. The possible stimulus to endocytosis is discussed. The endocytic response to inducers does not appear to be an all-or-none phenomenon and varies with the concentration of inducer. The stimulatory effect for protein-related inducers seems to be produced by a large number of stimulatory molecules acting upon a single cell and the magnitude of the response appears to be related to molecular size.     

Isocitrate dehydrogenase of Tetrahymena pyriformis

Summary We have studied the isocitrate dehydrogenase ofTetrahymena pyriformis. This enzyme is able to utilize both NAD and NADP, but kinetic studies suggest that the enzymatic activity with NAD is not of physiological significance.Some of the factors that might regulate the NADP-dependent isocitrate dehydrogenase were also studied. This enzyme has an absolute requirement for divalent cations; Mg²⁺ and Mn²⁺ will serve as cofactors but the latter is more effective than the former.It is known that this enzyme is subject to a concerted inhibition by oxaloacetate and glyoxylate. Either glyoxylate or oxaloacetate alone also are capable of inhibiting the enzyme although higher concentrations are required. We have found concerted inhibition also for the NAD-dependent isocitrate dehydrogenase from rat liver and yeast. The activity of theTetrahymena pyriformis enzyme is inhibited by NADPH. This inhibition is competitive with NADP. The Ki and Km values are, respectively, 23µ m and 18µ m.