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.     

THE FINE STRUCTURE OF ACOUSTIC GANGLIA IN THE RAT

Nerve cell bodies in the spiral and vestibular ganglia of the adult rat are surrounded by thin (about ten lamellae) myelin sheaths which differ in several respects from typical axonal myelin. In some instances lamellae surrounding perikarya appear as typical major dense lines, and in others as thin Schwann cell sheets in which cytoplasm persists. Discontinuities and irregularities appear in the structure of perikaryal myelin. Lamellae may terminate anywhere within the sheaths; they may bifurcate; they may reverse their direction; or they may merge with each other. The number of lamellae varies from one part of a sheath to another. In addition, the myelin of a single perikaryal sheath may receive contributions from more than one Schwann cell, which overlap and interleave with each other. The ganglion cells are of two types: those which are densely packed with the usual cytoplasmic organelles but have few neurofilaments (granular neurons), and those which exhibit large areas containing few organelles but have a high concentration of neurofilaments (filamented neurons). The latter cell type is ensheathed by myelin which is generally more compact that that surrounding the former. The formation and the physiologic significance of perikaryal myelin are discussed.     

THE FINE STRUCTURE OF THE TRANSITIONAL EPITHELIUM OF RAT URETER

The fine structure of the transitional epithelium of rat ureter has been studied in thin sections with the electron microscope, including some stained cytochemically to show nucleoside triphosphatase activity. The epithelium is three to four cells deep with cuboidal or columnar basal cells, intermediate cells, and superficial squamous cells. The basal cells are attached by half desmosomes, or attachment plates, on their basal membranes to a basement membrane which separates the epithelium from the lamina propria. Fine extracellular fibres, ca. 100 A in diameter, are to be found in the connective tissue layer immediately below the basement membrane of this epithelium. The plasma membranes of the basal and intermediate cells and the lateral and basal membranes of the squamous cells are deeply interdigitated, and nucleoside triphosphatase activity is associated with them. All the cells have a dense feltwork of tonofilaments which ramify throughout the cytoplasm. The existence of junctional complexes, comprising a zonula occludens, zonula adhaerens, and macula adhaerens or desmosome, between the lateral borders of the squamous cells is reported. It is suggested that this complex is the major obstacle to the free flow of water from the extracellular spaces into the hypertonic urine. The free luminal surface of the squamous cells and many cytoplasmic vesicles in these cells are bounded by an unusually thick plasma membrane. The three leaflets of this unit membrane are asymmetric, with the outer one about twice as thick as the innermost one. The vesicles and the plasma membrane maintain angular conformations which suggest the membrane to be unusually rigid. No nucleoside triphosphatase activity is associated with this membrane. Arguments are presented to support a suggestion that this thick plasma membrane is the morphological site of a passive permeability barrier to water flow across the cells, and that keratin may be included in the membrane structure. The possible origin of the thick plasma membrane in the Golgi complex is discussed. Bodies with heterogeneous contents, including characteristic hexagonally packed stacks of thick membranes, are described. It is suggested that these are "disposal units" for old or surplus thick membrane. A cell type is described, which forms only 0.1 to 0.5 per cent of the total cell population and contains bundles of tubular fibres or crystallites. Their origin and function are not known.     

THE FINE STRUCTURE OF EPENDYMA IN THE BRAIN OF THE RAT

The ciliated ependyma of the rat brain consists of a sheet of epithelial cells, the luminal surface of which is reflected over ciliary shafts and numerous evaginations of irregular dimensions. The relatively straight lateral portions of the plasmalemma of contiguous cells are fused at discrete sites to form five-layered junctions or zonulae occludentes which obliterate the intercellular space. These fusions occur usually at some distance below the free surface either independently or in continuity with a second intercellular junction, the zonula adhaerens. The luminal junction is usually formed by a zonula adhaerens or, occasionally, by a zonula occludens. The finely granular and filamentous cytoplasm contains supranuclear dense bodies, some of which are probably lysosomes and dense whorls of perinuclear filaments which send fascicles toward the lateral plasmalemma. The apical regions of the cytoplasm contain the basal body complexes of neighboring cilia. These complexes include a striated basal foot and short, non-striated rootlets emanating from the wall of each basal body. The rootlets end in a zone of granules about the proximal region of the basal body, adjacent to which may lie a striated mass of variable shape. All components of the basal body complex of adjacent cilia are independent of each other.     

FINE STRUCTURE CHANGES DURING FUNCTION OF THE DIGESTIVE GLAND OF VENUS'S-FLYTRAP

Changes in the structure of the digestive gland cells of Venus's-flytrap during the digestive process have been studied with light and electron microscopy. Large vacuolar lipid-protein inclusions break up and become smaller; however, they never completely disappear during the entire 7-10-day cycle. Dictyosomes in the resting digestive gland are associated with small, inconspicuous vesicles, whereas during the digestive cycle two types of prominent vesicles are observed on the peripheral tubules. Changes in plastid fine structure are complex and involve the disappearance of lipid globules and the tubular complex, followed by the formation of microtubules on the thylakoids and cisternae on the outer plastid membrane. Mitochondrial fine structure changes from the small cristae and light matrix of the resting state to large cristae and a very dense matrix representative of a change to a state where phosphorylation is tightly coupled to electron transport. Pronounced changes which occur in the cell envelope (cell wall and membrane taken together) are apparently associated with secretion of the digestive fluid. Numerous other changes are observed such as polysome formation, multivesicular body formation, mitochondria division, and changes which can be attributed in general to elevated cell activity.     

THE FINE STRUCTURE OF MITOSIS IN RAT THYMIC LYMPHOCYTES

The fine structure of rat thymic lymphocytes from early prophase to late telophase of mitosis is described, using material fixed at pH 7.3 either in 1 per cent OsO4 or in glutaraldehyde followed by 2 per cent OsO4. The structure of the centriolar complex of interphase thymocytes is analyzed and compared with that of centrioles during division. The appearance of daughter centrioles is the earliest clearly recognizable sign of prophase. Daughter centrioles probably retain a secondary relation to the primary centriole, while the latter appears to be related, both genetically and spatially, to the spindle apparatus. The nuclear envelope persists in recognizable form to help reconstitute the envelopes of the daughter nuclei. Ribosome bodies (dense aggregates of ribosomes) accumulate, beginning at late prophase, and are retained by the daughter cells. Cytokinesis proceeds by formation of a ribosome-free plate at the equator with a central plate of vesicles which may coalesce to form the new plasma membrane of the daughter cells. Stages in the formation of the midbody are illustrated.     

THE FINE STRUCTURE OF THE PIGMENT EPITHELIUM IN THE ALBINO RAT

In this report, particular attention is paid to the inclusion bodies found in the apical cytoplasm of the pigment epithelial cell. These bodies are of variable size and form. The smallest (0.4 µ diameter) consist of a granular matrix enclosed by a single membrane, and are similar to the lysosomes of hepatic cells. Larger inclusion bodies contain areas of lamellated material in addition to granular matrix. The largest particles seen (2 µ diameter) are almost entirely lamellar. These different forms seem closely related, for it is possible to find all transitional stages between the smallest and largest particles. The relationship between the lamellar inclusion bodies and the rod outer segments is discussed.     

THE FINE STRUCTURE OF OSTEOBLASTS IN THE METAPHYSIS OF THE TIBIA OF THE YOUNG RAT

The appearance of osteoblasts after fixation with OsO₄ is described in this paper. They have the basic structures found in other types of cells. The most striking feature is the array of rough-surfaced membranes of the endoplasmic reticulum; this feature is in keeping with the osteoblast's function of producing collagen as the bone grows. The sacs formed by these membranes probably represent the protein-containing granules described by other workers using the light microscope. They contain fine fibrillary material, and similar fibrils are to be found free in the cytoplasm. These fibrils could be tropocollagen units, although fibrils recognizable as collagen by their structure are found only outside the cell. The arrangement of the cell organelles does not seem to be related to the formation of collagen, but correlation of the fine structures of the cells with the histochemical and cytochemical findings in these cells reported by other workers leaves no doubt that they are directly concerned in the production of the organic matrix. It has not been possible to show that osteoblasts influence the passage of calcium or phosphate ions from the blood to the bone matrix.     

SOME OBSERVATIONS ON THE FINE STRUCTURE OF THE LUTEIN CELLS OF X-IRRADIATED RAT OVARY

The ovaries of 10- to 13-week-old rats were exteriorized and irradiated with sterilizing doses of X-rays. Following treatment, the animals entered a phase of constant vaginal cornification. Animals were killed 8 to 12 wk after the onset of this phase, and their ovaries were prepared for electron microscopy. Tissue was fixed in glutaraldehyde, postfixed in osmium tetroxide (Millonig's, phosphate-buffered), and embedded in Epon. Lutein cells from these ovaries were compared with those from sham-irradiated controls. The cytoplasm of lutein cells from experimental animals was characterized by an increase in the amount of agranular endoplasmic reticulum and by an increase in the number of mitochondria. These mitochondria are more variable in external form and often possess increased numbers of villiform cristae. Other features noted were a decrease in the amount of cytoplasmic lipid granules and an increase in cell size and surface irregularity. The significance of the morphological findings is discussed in relation to ovarian hormone production in animals sterilized by X-irradiation.     

THE ACTION OF EXCESS OF VITAMIN A ALCOHOL ON THE FINE STRUCTURE OF RAT DERMAL FIBROBLASTS

Rat dermal fibroblasts were grown as monolayers, and changes in the fine structure of the cells that occurred during 12 hr incubation in a medium containing protein and excess of retinol (vitamin A alcohol) were studied by electron microscopy. There is little change during the first 6 hr, although some of the nuclei have highly convoluted membranes. During the subsequent 3 hr, there is some disorganization of the mitochondrial cristae; the cisternae of the rough-surfaced endoplasmic reticulum diminish in number; and the amount of smooth membranous material and free ribosomes increases. There is a rapid decline in the respiratory activity of the cells after 6 hr exposure to the vitamin. It is concluded that the primary action of excess of retinol is to cause alterations in the membranes of the cells and that these alterations affect the functions of the mitochondria and endoplasmic reticulum.     

THE FINE STRUCTURE OF PROLIFERATING CELLS IN PRENEOPLASTIC RAT LIVERS DURING AZO-DYE CARCINOGENESIS

The continuous feeding of the carcinogenic aminoazo dye DAB to rats produces hyperbasophilic foci in the preneoplastic livers. After injections of thymidine-³H into the rats, such foci were isolated from the livers and studied by radioautography with the phase-contrast and electron microscopes. In these foci, the only cells found to be proliferating, as determined by the uptake of thymidine-³H into their nuclei, were a poorly differentiated type; well differentiated hepatocytes in the same regions were not labeled with the isotope. The labeled cells had an irregular cell outline and a high nucleocytoplasmic ratio; the cytoplasm had almost completely lost the specialized elements characteristic of hepatocytes; the irregular nuclei with prominent nucleoli, the altered mitochondria, and the increased free ribosomes noted in these cells are features which are characteristic of neoplastic cells induced by DAB. Thus, it seems likely that the hyperbasophilic foci represent the sites of extensive dedifferentiation of hepatocytes followed by rapid cellular proliferation, leading to neoplastic growth.     

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.     

CHANGES IN FINE STRUCTURE DURING SILK PROTEIN PRODUCTION IN THE AMPULLATE GLAND OF THE SPIDER ARANEUS SERICATUS

The ampullate silk gland of the spider, Araneus sericatus, produces the silk fiber for the scaffolding of the web. The fine structure of the various parts of the gland is described. The distal portion of the duct consist of a tube of epithelial cells which appear to secrete a substance which forms the tunica intima of the duct wall. At the proximal end of the duct there is a region of secretory cells. The epithelium of the sac portion contains five morphologically distinct types of granules. The bulk of the synthesis of silk occurs in the tail of the gland, and in this region only a single type of secretory droplet is seen in the epithelium. Protein synthesis can be stimulated by the injection of 1 mg/kg acetylcholine into the body fluids. 10 min after injection, much of the protein stored in the cytoplasm of the epithelial cells has been secreted into the lumen. 20 min after stimulation, the ergastoplasmic sacs form large whorls in the cytoplasm. Protein, similar in electron-opacity to protein found in the lumen, begins to form in that portion of the cytoplasm which is enclosed by the whorls. The limiting membrane of these droplets is formed by ergastoplasmic membranes which lose their ribosomes. No Golgi material has been found in these cells. Protein appears to be manufactured in the cytoplasm of the tail cells in a form which is ready for secretion.     

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.     

BIOSYNTHESIS OF S-ADENOSYL-L-METHIONINE IN THE RAT PINEAL GLAND

This investigation provides direct evidence that rat pineal gland contains ATP:L-methionine S-adenosyltransferase. The specific activity of the enzyme is about 25 times greater than that of hydroxyindole-O-methyltransferase, the unique marker enzyme in this tissue. The major product of the reaction catalyzed by ATP:L-methionine S-adenosyltransferase has been identified as S-adenosyl-methionine which can function as a methyl group donor for the formation of melatonin. The specific activity of pineal ATP:L-methionine S-adenosyltransferase is at least eight times greater than that of brain, and one-half that of the liver enzyme.     

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 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.     

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.