FINE STRUCTURE OF MEMBRANOUS AND MICROFIBRILLAR SYSTEMS IN THE CORTEX OF PARAMECIUM CAUDATUM

An electron microscope study of the cortex of Paramecium caudatum has revealed new details pertinent to several unresolved problems. The lateral boundaries of the alveoli do not regularly follow the crests of the polygonal ridges and thus their staining with silver cannot account for the external lattice seen by light microscopists. A granulo-fibrillar material is present, however, within the peaks of the ridges, which would account for the external lattice if so stained. Perforations are present between adjacent alveoli which make the whole mosaic of alveolar sacs within the cell's cortex continuous—both the membranes and the lumen. A microfibrillar system exhibiting a cross-striated pattern lies in the superficial cortex. These bands are inserted at their ends in the epiplasm and have a fine structure and arrangement suggesting a muscular function. The infraciliary lattice is a branching system of fibers with electron-opaque posts at the center of each branching locus. This system is distinct from the striated bands in morphology and in space. The epiplasm is discontinuous along the crests of the ridges, which may account for the pellicles' disposition to tear along these lines. A three-dimensional drawing is presented to show the interrelationships between the above membranous and microfibrillar systems.     

THE FINE STRUCTURE OF NEURONS

1. Thin sections of representative neurons from intramural, sympathetic and dorsal root ganglia, medulla oblongata, and cerebellar cortex were studied with the aid of the electron microscope. 2. The Nissl substance of these neurons consists of masses of endoplasmic reticulum showing various degrees of orientation; upon and between the cisternae, tubules, and vesicles of the reticulum lie clusters of punctate granules, 10 to 30 mµ in diameter. 3. A second system of membranes can be distinguished from the endoplasmic reticulum of the Nissl bodies by shallower and more tightly packed cisternae and by absence of granules. Intermediate forms between the two membranous systems have been found. 4. The cytoplasm between Nissl bodies contains numerous mitochondria, rounded lipid inclusions, and fine filaments.     

THE FINE STRUCTURE OF THE PURKINJE CELL

This paper describes the fine structure of the Purkinje cell of the rat cerebellum after fixation by perfusion with 1 per cent buffered osmium tetroxide. Structures described include a large Golgi apparatus, abundant Nissl substance, mitochondria, multivesicular bodies, osmiophilic granules, axodendritic and axosomatic synapses, the nucleus, the nucleolus, and the nucleolar body. A new and possibly unique relationship between mitochondria and subsurface cisterns is described. Possible functional correlations are discussed.     

THE FINE STRUCTURE OF ELASTIC FIBERS

The fine structure of developing elastic fibers in bovine ligamentum nuchae and rat flexor digital tendon was examined. Elastic fibers were found to contain two distinct morphologic components in sections stained with uranyl acetate and lead. These components are 100 A fibrils and a central, almost amorphous nonstaining area. During development, the first identifiable elastic fibers are composed of aggregates of fine fibrils approximately 100 A in diameter. With advancing age, somewhat amorphous regions appear surrounded by these fibrils. These regions increase in prominence until in mature elastic fibers they are the predominant structure surrounded by a mantle of 100 A fibrils. Specific staining characteristics for each of the two components of the elastic fiber as well as for the collagen fibrils in these tissues can be demonstrated after staining with lead, uranyl acetate, or phosphotungstic acid. The 100 A fibrils stain with both uranyl acetate and lead, whereas the central regions of the elastic fibers stain only with phosphotungstic acid. Collagen fibrils stain with uranyl acetate or phosphotungstic acid, but not with lead. These staining reactions imply either a chemical or an organizational difference in these structures. The significance and possible nature of the two morphologic components of the elastic fiber remain to be elucidated.     

THE FINE STRUCTURE OF GREEN BACTERIA

The fine structure of several strains of green bacteria belonging to the genus Chlorobium has been studied in thin sections with the electron microscope. In addition to having general cytological features typical of Gram-negative bacteria, the cells of these organisms always contain membranous mesosomal elements, connected with the cytoplasmic membrane, and an elaborate system of isolated cortical vesicles, some 300 to 400 A wide and 1000 to 1500 A long. The latter structures, chlorobium vesicles, have been isolated in a partly purified state by differential centrifugation of cell-free extracts. They are associated with a centrifugal fraction that has a very high specific chlorophyll content. In all probability, therefore, the chlorobium vesicles are the site of the photosynthetic apparatus of green bacteria.     

THE FINE STRUCTURE OF ANNULATE LAMELLAE

Certain lamellar structures have been described from snail (Otala) and clam (Spisula) oocytes, the acinar cells of amphibian (Ambystoma) pancreas, and from rat spermatids. These structures are alike in possessing numerous rings or annuli, resembling those in the nuclear membrane. Thus the name "annulate lamellae" has been proposed for them. It is suggested that they may function in the transfer of specificities from nucleus to cytoplasm.     

THE FINE STRUCTURE OF GIARDIA MURIS

Giardia is a noninvasive intestinal zooflagellate. This electron microscope study demonstrates the fine structure of the trophozoite of Giardia muris in the lumen of the duodenum of the mouse as it appears after combined glutaraldehyde and acrolein fixation and osmium tetroxide postfixation. Giardia muris is of teardrop shape, rounded anteriorly, with a convex dorsal surface and a concave ventral one. The anterior two-thirds of the ventral surface is modified to form an adhesive disc. The adhesive disc is divided into 2 lobes whose medial surfaces form the median groove. The marginal grooves are the spaces between the lateral crests of the adhesive disc and a protruding portion of the peripheral cytoplasm. The organism has 2 nuclei, 1 dorsal to each lobe of the adhesive disc. Between the anterior poles of the nuclei, basal bodies give rise to 8 paired flagella. The median body, unique to Giardia, is situated between the posterior poles of the nuclei. The cytoplasm contains 300-A granules that resemble particulate glycogen, 150- to 200-A granules that resemble ribosomes, and fusiform clefts. The dorsal portion of the cell periphery is occupied by a linear array of flattened vacuoles, some of which contain clusters of dense particles. The ventrolateral cytoplasm is composed of regularly packed coarse and fine filaments which extend as a striated flange around the adhesive disc. The adhesive disc is composed of a layer of microtubules which are joined to the cytoplasm by regularly spaced fibrous ribbons. The plasma membrane covers the ventral and lateral surfaces of the disc. The median body consists of an oval aggregate of curved microtubules. Microtubules extend ventrally from the median body to lie alongside the caudal flagella. The intracytoplasmic portions of the caudal, lateral, and anterior flagella course considerable distances, accompanied by hollow filaments adjacent to their outer doublets. The intracytoplasmic portions of the anterior flagella are accompanied also by finely granular rodlike bodies. No structures identifiable as mitochondria, smooth endoplasmic reticulum, the Golgi complex, lysosomes, or axostyles are recognized.     

FINE STRUCTURE OF THE CILIA OF ROTIFERS

The fine structure of the coronal cilia of the rotifer Philodina citrina has been studied in detail. Specimens were fixed with OsO₄ and embedded in butyl—methyl methacrylate, Epon 812, or Vestopal and sectioned with a Porter-Blum microtome. The details of structure of the rootlets, basal bodies, basal plates, and free cilia are described. The general structure of the rotifer ciliary apparatus conforms well to that established for other species. One of the main observations is the difference in structure of the peripheral filaments in the opposing halves of a cross-section of the free cilium. Also, in longitudinal sections evidence is offered for the existence of a helical structure in the peripheral filaments.     

THE FINE STRUCTURE OF DIPLOCOCCUS PNEUMONIAE

The fine structure of an unencapsulated strain of Diplococcus pneumoniae is described. A striking feature of these bacteria is an intracytoplasmic membrane system which appears to be an extension of septa of dividing bacteria. The possible function of these structures and their relationship to the plasma membrane and other types of intracytoplasmic membranes found in pneumococcus is discussed.     

THE FINE STRUCTURE OF RHODOSPIRILLUM RUBRUM

The fine structure of Rhodospirillum rubrum grown under a series of defined conditions has been examined in thin sections prepared by the methods of Ryter and Kellenberger. In cells grown anaerobically at different light intensities, the abundance of 500 A membrane-bounded vesicles in the cytoplasm is inversely related to light intensity, and directly related to cellular chlorophyll content. When the chlorophyll content of the cell is low, the vesicles are exclusively peripheral in location; they extend more deeply into the cytoplasm when the chlorophyll content is high. Typical vesicles also occur, though rarely, in cells grown aerobically in the dark, which have a negligible chlorophyll content. When synthesis of the photosynthetic pigment system is induced in a population of aerobically grown cells by incubation under semianaerobic conditions in the dark, the vesicles become increasingly abundant with increasing cellular chlorophyll content, and the cells eventually acquire the cytoplasmic structure that is characteristic of cells growing anaerobically at a high light intensity. Poststaining with lead hydroxide reveals that the membranes surrounding the 500 A vesicles are indistinguishable in structure from the cytoplasmic membrane, and continuous with it in some areas of the sections. The bearing of these observations on current notions concerning the organization of the bacterial photosynthetic apparatus is discussed.     

THE FINE STRUCTURE OF BONE CELLS

An electron microscopic study of Araldite-embedded, undecalcified human woven and chick lamellar bone is presented. The fine structure of the cells of bone in their normal milieu is described. Active osteoblasts possess abundant granular endoplasmic reticulum, numerous small vesicles, and a few secretion droplets. Their long cytoplasmic processes penetrate the osteoid. The transition of osteoblasts into osteoid osteocytes and then into osteocytes is traced and found to involve a progressive reduction of cytoplasmic organelles. Adjoining the osteocytes and their processes is a layer of amorphous material which is interposed between the cell surfaces and the bone walls of their respective cavities. Osteoclasts contain numerous non-membrane-associated ribosomes, abundant mitochondria, and little granular endoplasmic reticulum, thus differing markedly from other bone cells. The brush border is a complex of cytoplasmic processes adjacent to a resorption zone in bone. No unmineralized collagen is seen at resorption sites and it appears that collagen is removed before or at the time of mineral solution. All bone surfaces are covered by cells, some of which lack distinctive qualities and are designated endosteal lining cells. The structure of osteoid, bone, and early mineralization sites is illustrated and discussed.     

FINE STRUCTURE OF THE OCTOPUS RETINA

The fine structure of the visual and the supporting cells and of the blood capillaries in the octopus retina is described. Lamellated structures contained in the proximal segment of the visual cell consist of compact arrays of dense membranes each of which is quintuple-layered and divides at its margins into two thinner sheets or membranes which are connected directly with the agranular or granular endoplasmic reticulum. Proximal to the deeper extremities of the rhabdomeres, the lateral plasma membranes of two adjoining visual cells contact each other forming a quintuple-layered compound membrane, which results in occlusion of the intercellular space. The central layer of the compound membrane is of high density, so that the membrane, as a whole, appears to be a single thick layer at low magnifications. The supporting cells are connected with the neighboring visual cells by two types of junctions. Long slender processes extend from the supporting cells to the surface of the retina through narrow spaces among the distal segments of the visual cells. The capillary endothelial cells are characterized by luminal surfaces irregularly contoured and by lateral surfaces elaborately interdigitated. The functional significance of the close contact between adjoining visual cells is discussed.     

THE FINE STRUCTURE OF THE MID-BODY OF THE RAT ERYTHROBLAST

The development of the mid-body has been studied in mitotic erythroblasts of the rat bone marrow by means of thin sections examined with the electron microscope. A differentiated region on the continuous spindle fibers, consisting of a localized increase in density, is observed at the equatorial plane. The mid-body seems to develop by the aggregation of such denser lengths of spindle fiber. Its appearance precedes that of the cleavage furrow. A plate-like arrangement of fibrillary material lies transversely across the telophase intercellular bridge. Later, this material becomes amorphous and assumes the form of a dense ring closely applied to a ridge in the plasma membrane encircling the middle of the bridge. Although the mid-body forms in association with the spindle fibers, it is a structurally distinct part, and the changes which it undergoes are not shared by the rest of the bundle of continuous fibers.     

FINE STRUCTURE OF THE NERVOUS SYSTEM OF HYDRA

Fine structural details of the cells and processes of the hydranenous system are reported in this paper. Ganglion cells aresmall bipolar or multipolar cells situated above the muscularprocesses of epitheliomusiular cells. An elaborate Colgi apparatusconsisting of parallel lamellae and small and large vesiclesis present in these cells. Some cells are poor in ribosomeswhile others contain numerous free ribosomes. In the ribosome-richcells, small membranous microtubules originating from the nuclearenvelope extend into the cytoplasm and neurites. The neuritesalso contain vesicles and mitochondria and terminate at thebases of cnidoblasts and on the muscular processes of epitheliomuscularcells. Specialized synapses were not observed. A second cell type contains many membrane-bounded dense granules,1000 A in diameter, and these are considered to be neurosecretorycells. Neurosecretory granules on cnidoblasts and epitheliomuscularcells. Sensory cells are small elongated cells originate inthe Golgi apparatus and are abundant in neurites which alsoterminate situated between the apical surfaces of epithelialand digestive cells. These cells are characterized by an apicalspecialization which appears to be a modified cilium. Neurosensorycells were also observed. The intimate connection of the nervoussystem with cnidoblasts suggests a role in nematocyst discharge.The finding of neurosecretory material supports the hypothesisthat the neural control of regeneration in hydra is regulatedby material released at nerve endings.     

FINE STRUCTURE OF THE EYE OF A CHAETOGNATH

Electron microscopy reveals a star-like pigment cell at the center of the eye of the arrow-worm, Sagitta scrippsae. Between the arms of the pigment cell are clusters of photoreceptor cell processes, each process consisting of: (1) a tubular segment containing longitudinally arranged microtubules about 500 A in diameter and 20 µ in length; (2) a remarkable conical body, composed of cords and large granules, situated at the base of the tubular segment; and (3) a connecting piece which, like that of rods and cones, connects the process with the sensory cell proper and through which runs a fibrillar apparatus consisting of nine peripheral double tubules. Beneath the connecting piece lies a typical centriole with a striated rootlet. The receptor cell process is deeply recessed into the sensory cell which may possess a corona of microvilli at its inner surface. A nerve fiber arises from the outer end of the cell and passes into the optic nerve. Additional features are some supporting cells, an external layer of flattened epithelial cells, and an over-all investment of basement membrane and thick fibrous capsule. The fine structure and function of these elements of the eye are discussed in relation to earlier studies with the light microscope. The ciliary nature of the photoreceptor cell process in S. scrippsae points to a probable evolutionary relationship of chaetognaths to echinoderms and chordates.