Studies on the metabolism of the Protozoa. III. Metabolism of the ciliate Tetrahymena pyriformis (Glaucoma piriformis).
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1. Cell-free extracts from Epidinium ecaudatum (Crawley) hydrolysed the three hemicellulose fractions of pasture plants, but at different rates. 2. All of the constituent monosaccharides are released from the hemicellulose fractions, galactose and uronic acids being liberated at much slower rates than pentoses. 3. An arabinofuranosidase, which removes arabinose from highly branched arabinoxylan before the xylan chain can be hydrolysed, was isolated free from other pentosanases. 4. A xylanase hydrolysing xylan (by random cleavage) and xylodextrins of degree of polymerization (D.P.) > 3 to xylotriose and xylobiose was isolated free from other pentosanases. 5. A separate xylodextrinase hydrolysing (by random cleavage) xylodextrins of D.P. > 2 to xylobiose and xylose was also obtained; this enzyme did not hydrolyse xylan or xylobiose and the original extracts themselves possessed very weak xylobiase activity. 6. The epidinial extracts hydrolysed laminaribiose, laminarin, lichenin and cellodextrins of D.P. < 7 rapidly, cellobiose and gentiobiose slowly but cellulose not at all. 7. Polysaccharide glucose associated with plant linear B hemicellulose was liberated with cellobiose and possibly laminaribiose as intermediates. 8. The cellodextrinase hydrolysed cellopentaose initially to cellobiose plus cellotriose and is a distinctly different enzyme from the xylanase and xylodextrinase. 9. Extracts from Entodinium species and Eremoplastron bovis also hydrolysed all three types of plant hemicellose.
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The electron microscope was used to study the structure and three dimensional relationships of the components of the body cortex in thin sections of Paramecium multimicronucleatum. Micrographs of sections show that the cortex is covered externally by two closely apposed membranes (together approximately 250 A thick) constituting the pellicle. Beneath the pellicle the surface of the animal is molded into ridges that form a polygonal ridgework with depressed centers. It is these ridges that give the surface of the organism its characteristic configuration and correspond to the outer fibrillar system of the light microscope image. The outer ends of the trichocysts with their hood-shaped caps are located in the centers of the anterior and posterior ridges of each polygon. The cilia extend singly from the depressed centers of the surface polygons. Each cilium shows two axial filaments with 9 peripheral and parallel filaments embedded in a matrix and the whole surrouned by a thin ciliary membrane. The 9 peripheral filaments are double and these are evenly spaced in a circle around the central pair. The ciliary membrane is continuous with the outer member of the pellicular membrane, whereas the plasma membrane is continuous with the inner member of the pellicular membrane. At the level of the plasma membrane the proximal end of the cilium is continuous with its tube-shaped basal body or kinetosome. The peripheral filaments of the cilium, together with the material of cortical matrix which tends to condense around them, form the sheath of the basal body. The kinetodesma connecting the ciliary kinetosomes (inner fibrillar system of the light microscopist) is composed of a number of discrete fibrils which overlap in a shingle-like fashion. Each striated kinetosomal fibril originates from a ciliary kinetosome and runs parallel to other kinetosomal fibrils arising from posterior kinetosomes of a particular meridional array. Sections at the level of the ciliary kinetosomes reveal an additional fiber system, the infraciliary lattice system, which is separate and distinct from the kinetodesmal system. This system consists of a fibrous network of irregular polygons and runs roughly parallel to the surface of the animal. Mitochondria have a fine structure similar in general features to that described for a number of mammalian cell types, but different in certain details. The structures corresponding to cristae mitochondriales appear as finger-like projections or microvilli extending into the matrix of the organelle from the inner membrane of the paired mitochondrial membrane. The cortical cytoplasm contains also a particulate component and a system of vesicles respectively comparable to the nucleoprotein particles and to the endoplasmic reticulum described in various metazoan cell types. An accessory kinetosome has been observed in oblique sections of a number of non-dividing specimens slightly removed from the ciliary kinetosome and on the same meridional line as the cilia and trichocysts. Its position corresponds to the location of the kinetosome of the newly formed cilium in animals selected as being in the approaching fission stage of the life cycle.
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The macronuclei of synchronously dividing mass cultures of Tetrahymena pyriformis (strain WH(6)) were examined with the electron microscope for changes during two division cycles. Samples were prepared at 30-minute intervals for a period of 8(1/2) hours which included the time required to induce synchrony by five heat shocks (4(1/2) hours). The interphase macronucleus contains peripheral, crescent-shaped nucleoli and evenly distributed chromatin bodies. Centrally located RNA bodies, composed of fibers, appear 1 to 2 hours following the initial heat shock. They are completely destroyed with ribonuclease whereas the nucleoli are only partially so. Following the third heat shock the RNA bodies move to the periphery and disintegrate; the nucleoli aggregate and form blebs which protrude into the cytoplasm where they appear to pinch off and may contribute to the cytoplasmic ribonucleic acid. Cytokinesis does not occur at this time. Instead the nuclear events are repeated during the 4th and 5th hours, even though the heat shocks are terminated at 4(1/2) hours. Cytokinesis takes place at about 6 hours. The second division occurs about 2(1/2) hours later during which all the macronuclear events noted above are repeated.
1. The functional unity of cirri and membranelles can result structurally only from extensions of the ciliary membrane. 2. The pellicle is composed of an outer pellicular membrane and an inner cytoplasmic membrane. 3. The ciliary rootlets are composed of numerous filaments 120 A in diameter with central areas of low density. They have no periodic structure. 4. The ciliary membrane is a double-layered structure continuous with the pellicular membrane. The cilia show the typical arrangement of nine double, peripheral and two single, central fibrils. All fibrils pass into the basal region, the peripheral ones joining with the rootlet filaments, while the central fibrils from the extreme proximal position of the basal region turn back toward the pellicle and appear to unite just beneath the cytoplasmic membrane. 5. The cilia (300 mmicro diameter) taper at their tips to a diameter at least as small as 50 mmicro. At a diameter of about 150 mmicro, the fibrils begin to show a reduction in number. 6. The central ciliary fibrils may determine the possible directions of ciliary beat. These fibrils show an intrafibrillar structure in their basal portion, which involves regularly spaced 40 A granules. 7. These observations on Euplotes, together with the other evidence cited, are consistent with the hypothesis that ciliary motion is produced by the contraction of the peripheral fibrils, while the central fibrils perhaps determine the plane in which the cilia can bend.
1. Structurally the "sensory bristles" in Euplotes patella are typical cilia, but no ciliary rootlets connect their bases. 2. The "neuromotor fibrils" are composed of filaments 21 mmicro in diameter. At the point of junction of the filaments with the peripheral ciliary fibrils a granular structure 65 to 90 mmicro in diameter is seen which has dense central and peripheral zones separated by a less dense layer. Information on the interconnection of organelles is expanded. 3. A system of subpellicular fibrils is described. The external fibrillar system described by others could not be found. 4. The motorium is shown to be a mass of intertwining rootlet filaments. 5. The micronucleus is shown to have a spongy, dense material in a less dense material, all of which is surrounded by a double-layered membrane. 6. The double-layered macronuclear membrane contains annuli whose outside diameter is 70 mmicro; the macronuclear bodies are sometimes closely applied to the membrane. In the macronuclear reorganization bands, the solution plane is a fine network, while the reconstruction plane is devoid of structure at the level of resolution observed. 7. The mitochondria are composed of tubules, only occasionally oriented, usually embedded in a surrounding material of lower density. 8. Microbodies whose diameters are 250 to 350 mmicro are frequently observed in close association with mitochondrial surfaces. 9. The food vacuoles, contractile vacuoles, and ciliary vacuoles are bounded by single-layered membranes. In the food vacuoles, the bacteria are surrounded by membranes individually or in small groups. 10. Cytoplasmic rods localized in the oral region, and cytoplasmic granules dispersed at random, are described. No typical ergastoplasm, endoplasmic reticulum, or Golgi material was observed.