THE CYTOPLASMIC FINE STRUCTURE OF THE DIATOM, NITZSCHIA PALEA

The cytoplasmic fine structure of the motile, pennate diatom, Nitzschia palea was studied in thin sections viewed in the electron microscope. The cells were fixed in OsO₄, embedded in methacrylate, and immersed in 10 per cent hydrofluoric acid (HF) for 36 to 40 hours to remove the siliceous cell wall prior to sectioning. The HF treatment did not cause any obvious cytoplasmic damage. The dictyosome complex is perinuclear, and located only in the central cytoplasm. Mitochondria are sparse in the central cytoplasm, but abundant in the peripheral cytoplasm, and fill many of the transvacuolar cytoplasmic strands. Characteristic, amorphous oil bodies fill certain cytoplasmic strands and probably are not leucosin. The pyrenoid appears to be membrane limited, and oil droplets are found adjacent to the pyrenoid. The pyrenoid of another diatom, Cymbella affinis, is also membrane-limited. The membrane limiting the pyrenoid may be a composite of the terminal portions of chloroplast discs, facilitating rapid movement of photosynthate into the pyrenoid matrix, where the characteristic oil droplets may be formed. Carinal fibrils are found singly in each carinal pore, and may be involved in the locomotion of Nitzschia palea.     

THE PRIMARY ROOT EPIDERMIS OF PANICUM VIRGATUM L. I. ONTOGENY AND FINE STRUCTURE OF THE EPIDERMAL CYTOPLASMIC INCLUSIONS

The ontogeny of large, globular, epidermal cytoplasmic inclusions (ECI) in P. virgatum roots was studied at the ultrastructural level. These ECI were seen to originate in meristematic cells as small electron translucent vesicles. Subsequently, the ECI, which appeared to be temporary storage sites, were seen to enlarge and increase in density by accumulating masses of a granular matrix as well as some small vesicular inclusions. In the zone of elongation, as the epidermal cells matured, the ECI within each cell gradually fused and the contents were lost. The pattern of the ontogeny of the ECI in the growing epidermal cells was consistent with the presence of cells of different physiologies in the zone of cell elongation of these roots.     

NANOSTRUCTURE OF THE DIATOM FRUSTULE AS REVEALED BY ATOMIC FORCE AND SCANNING ELECTRON MICROSCOPY

The cell wall (frustule) of the freshwater diatom Pinnularia viridis (Nitzsch) Ehrenberg is composed of an assembly of highly silicified components and associated organic layers. We used atomic force microscopy (AFM) to investigate the nanostructure and relationship between the outermost surface organics and the siliceous frustule components of live diatoms under natural hydrated conditions. Contact mode AFM imaging revealed that the walls were coated in a thick mucilaginous material that was interrupted only in the vicinity of the raphe fissure. Analysis of this mucilage by force mode AFM demonstrated it to be a nonadhesive, soft, and compressible material. Application of greater force to the sample during repeated scanning enabled the mucilage to be swept from the hard underlying siliceous components and piled into columns on either side of the scan area by the scanning action of the tip. The mucilage columns remained intact for several hours without dissolving or settling back onto the cleaned valve surface, thereby revealing a cohesiveness that suggested a degree of cross-linking. The hard silicified surfaces of the diatom frustule appeared to be relatively smooth when living cells were imaged by AFM or when field-emission SEM was used to image chemically cleaned walls. AFM analysis of P. viridis frustules cleaved in cross-section revealed the nanostructure of the valve silica to be composed of a conglomerate of packed silica spheres that were 44.8 ± 0.7 nm in diameter. The silica spheres that comprised the girdle band biosilica were 40.3 ± 0.8 nm in diameter. Analysis of another heavily silicified diatom, Hantzschia amphioxys (Ehrenberg) Grunow, showed that the valve biosilica was composed of packed silica spheres that were 37.1 ± 1.4 nm and that silica particles from the girdle bands were 38.1 ± 0.5 nm. These results showed little variation in the size range of the silica particles within a particular frustule component (valve or girdle band), but there may be differences in particle size between these components within a diatom frustule and significant differences are found between species.     

OBSERVATIONS ON THE STRUCTURE OF SOME FORMS OF GOMPHONEMA PARVULUM KÜTZ. III. FRUSTULE FORMATION1

Gomphonema parvulum Kütz. was investigated by electron microscopy for details of frustule formation. An expansion of the cell along the pervalvar plane occurs prior to cell division. After nuclear division the organelles are, separated into 2 entities, either by division or by dispersion. The cell divides into 2 halves by the invagination of the plasmalemma which is derived from Golgi vesicular activity. When cytoplasmic cleavage, is complete, the Golgi actively produces electronlucent vesicles which collect and coalesce beneath the. plasmalemma to form the silicalemma around the silicon deposition vesicle. The endoplasmic reticulum is also closely associated with this vesicular activity. The vesicle gradually expands and becomes extremely electron dense as silica is deposited within it—first in the region, followed by the mantle edge. When the valve is mature, Golgi vesicles collect and fuse to form the silicalemma of the first girdle band. The first girdle band becomes aligned against the mantle edge on completion, by the “sloughing off” of the external silicalemma and plasmalemma. The second and third bands are formed, individually in a similar manner. Separation of the 2 daughter cells commences at the apical pole and progresses to the basal pole. The plasmalemma and external silicalemma are “sloughed off” so that the 2 cells can separate. The inner segment of the silicalemma becomes the new plasmalemma of the daughter cell.     

CYTOPLASMIC FINE STRUCTURE OF SCIARA SALIVARY GLANDS : I. Secretion

Cells from the anterior segment of the salivary glands of Sciara coprophila were found to synthesize and secrete into the gland lumen three morphologically distinct types of granule: 1) A large, electron-lucid granule, up to 1 µ in diameter, staining only faintly with pH 2 fast green and the PAS reaction; 2) an ellipsoid granule of moderate density, strongly fast green and PAS positive; and 3) a small spherical granule of high electron density. The cells contained numerous Golgi areas, up to an estimated 8,000 per cell. Evidence is presented for the transfer of material from the endoplasmic reticulum to the Golgi areas via small vesicles. Three types of Golgi areas were distinguishable, each containing intercisternal material resembling one of the three types of secretion granule. Patterns of secretion granule synthesis varied with the developmental stage of the larva as determined by counts of eye spots in the eye anlage. Lucid granules were most abundant in the youngest larvae, and decreased in abundance as larvae grew older, becoming virtually absent in prepupae. The small, dense granules were present in all gland cells, but became more prevalent in older larvae and prepupae. Ellipsoid granules were only occasionally present, and were independent of larval stage. It is suggested that lucid granules are digestive in function, since their abundance correlates with feeding patterns. Other granules may produce the external slime coating of the larvae, and also the mucoprotein component of the pupal cocoon.     

STUDIES ON THE BIOCHEMISTRY AND FINE STRUCTURE OF SILICA SHELL FORMATION IN DIATOMS. II. THE STRUCTURE OF THE CELL WALL OF NAVICULA PELLICULOSA (BRÉB.) HILSE

The cell wall of the freshwater diatom Navicula pelliculosa (Bréb.) Hilse is composed of the silica shell and an organic skin which surrounds it. Isolated skins can be prepared by first removing the contents of the cell by mechanical shaking, followed by a posttreatment of these isolated cell walls with HF vapor to remove the silica shell. T h e skins can also be seen in sections, particularly well after the silica shell has been removed B y H. F; vapor. The origin and morphological composition of the shin in N. pelliculosa are not yet completely ascertaincd. As parts of the cell wn11, both the silica shell and the skin are extracellularly located. The growth of the silica shell, however, occurs intracellularly inside a vesicle delimited by a triple-layered membrane, the silicalemma. This membrane or secondary excreted organic material or both in various proportions may compose the skin.     

OCCURRENCE AND FINE STRUCTURE OF SUBCORTICAL CYTOPLASMIC ISLETS IN FERTILIZED EGGS OF CYNOPS PYRRHOGASTER

Fertilized and unfertilized eggs of Cynops pyrrhogaster were examined by light and electron microscopy. In fertilized eggs that have just been laid, there are numerous small cytoplasmic patches free of granules in the pigmented layer of the animal hemisphere. Many of these granule-free cytoplasmic islets gradually grow out subcortically from the pigmented layer and fuse to form a subcortical layer of yolk-free cytoplasm of varying thickness by the time of the first cleavage division. The cytoplasmic islets are present in 100% of the fertilized eggs, but not in unfertilized eggs. Electron microscopic observations showed that the cytoplasmic islets contain tubules and that development of a complex system of cortical tubules constitutes the basis of the early growth of the cytoplasmic islets. The cortical tubules are transient structures and are no longer observable a few hours after the eggs are laid. These phenomena are considered to be a response of the egg to the fertilization stimulus.     

FURTHER OBSERVATIONS AND SOME COMMENTS ON THE FINE STRUCTURE OF THE CENTRIC DIATOM AULACOSEIRA ISLANDICA (BACILLARIOPHYCEAE)1

Using light and electron microscopy, the diatom species Aulacoseira islandica (O. Müll.) Sim. was examined with special emphasis on the following characteristics: structure of the valve areolae, heterovalvy, and distribution of the rimoportulae. The mantle and valve face areolae were pores containing volate occlusions. However, observations only using transmission electron microscopy may result in an incomplete interpretation because of the fragility of the dissected system of volae. Relief valves with a stepped mantle and intaglio valves with a plain mantle occurred. Another form of heterovalvy resulted from the formation of separation valves. Linking valves had spatulate spines while separation valves bore tapering spines. In Aulacoseira, the rimoportulae usually occurred near the “Ringleiste.” The presence of several rimoportulae on the mantle was one of the most striking features in Aulacoseira islandica.     

THE FINE STRUCTURE OF CAPILLARIES AND SMALL ARTERIES

Details of capillary endothelia of the mammalian heart are described and compared with capillaries of other organs and tissues. Continuous invagination and pinching off of the plasma membrane to form small vesicles which move across the cytoplasm are suggested as constituting a means of active and selective transmission through capillary walls (12). This might be designated as cytopempsis (transmission by cell). The fine structure of the different layers in the walls of small heart arteries is demonstrated. Endothelial protrusions extend through windows of the elestica interna to make direct contact with smooth muscle plasma membranes. The elastica interna appears to vary greatly in both thickness and density, and probably restricts filtration, diffusion, and osmosis to such an extent that windows and the transport mechanisms described (cytopempsis) are necessary for the functional integrity of the smooth muscle layer. The contractile material consists of very fine, poorly oriented filaments.     

OBSERVATIONS ON THE FINE STRUCTURE OF OEDOGONIUM. II. THE SPERMATOZOID OF O. CARDIACUM

Hoffman , L. R., and Irene Manton . (U. Leeds, England.) Observations on the fine structure of Oedogonium. II. The spermatozoid of O. cardiacum. Amer. Jour. Bot. 50(5): 455–463. Illus. 1963.—Salient features of the fine structure of the spermatozoid of Oedogonium cardiacum are described and illustrated as they appear in whole mounts and in sections. There is a close resemblance to the zoospore of the same species (Hoffman and Manton, 1962) though the gamete is smaller and in some respects simpler. The flagella, though similar in length to those on the zoospore, are fewer (ca. 30 instead of ca. 120 per cell). The construction of the flagellar ring is similar though there is less mechanical material associated with the flagellar bases in the gamete. Compound “roots” alternating with the flagellar bases are identical in structure and relative position in both types of motile cells; there is no direct connection with the nucleus. Other details of resemblance and difference between the spermatozoid and the zoospore are discussed.     

PARTICIPATION OF THE CYTOPLASMIC MEMBRANE IN THE GROWTH AND SPORE FORMATION OF BACILLI

A polyester embedding technique was used to study the early stages of spore formation in members of the genus Bacillus in order to investigate further the origin and nature of the initial spore septum and the resulting forespore envelope. Whereas previously, with a methacrylate procedure, this layer had appeared to be continuous with the cell wall, this study reveals it as a double layer of cytoplasmic membrane. Perisporal, membranous organelles connected both to the developing forspore envelope and to the cytoplasmic membrane were encountered in the four species studied. Similar organelles were prominent during growth at the sites of transverse septa formation. These were connected to, or continuous with, the cytoplasmic membrane and often adherent to the chromatin bodies of the dividing bacilli.     

THE FINE STRUCTURE OF Streptomyces coelicolor : II. The Nuclear Material

Colonies and spore suspensions of Streptomyces coelicolor were fixed for electron microscopy by the method of Kellenberger, Ryter, and Séchaud (1958). In thin sections the nuclear regions have a lower average density than the cytoplasm and the outlines of these regions correspond well with the profiles of the chromatinic bodies observed with the light microscope. The nuclear regions contain fibrils, about 5 mµ in diameter. In contrast, after fixation by the method of Palade (1952) the nuclear material is coagulated into irregular dense masses and tubular structures about 20 mµ in diameter, lying in a nuclear "vacuole." The significance of these observations is discussed in relation to the observations of other workers on the fine structure of the nuclear material of other bacteria and the chromosomes of higher cells.     

ON THE FINE STRUCTURE AND COMPOSITION OF THE NUCLEAR ENVELOPE

Nuclei from nearly ripe eggs of Rana pipiens were isolated and cleaned in 0.1 M KCl. The whole nucleus was then digested to various degrees with ribonuclease or trypsin, followed by washing and fixation in either osmium tetroxide or potassium permanganate. The nuclear envelope was dissected off, placed on a grid, air dried, and compared with undigested controls in the electron microscope. Some envelopes were dehydrated, embedded in methacrylate, and sectioned. Annuli around "pores" are composed of a substance or substances, at least partially fibrillar, which is preserved by osmium but lost during permanganate fixation. Material within the "pores" is also preserved by osmium but partially lost after permanganate. No evidence of granules or tubules in the annuli was found in air dried mounts although a granular appearance could be seen in tangentially oriented thin sections. Thin sections of isolated envelopes give evidence of diffuse material within the "pores" as well as a more condensed diaphragm across their waists. In whole mounts of the envelope the total density within "pores" is relatively constant from "pore" to "pore." All material within "pores," including the condensed diaphragm, is removable by trypsin digestion. Wispy material from the "pore" structure projects into the nucleus and annular material extends into the cytoplasm. Both annular and diaphragm materials remain with the envelope when it is isolated and are thus considered a part of its structure, not merely evidences of material passing through. There is no evidence of ribonuclease-removable material in any part of the "pore" complex.     

OBSERVATIONS ON THE FINE STRUCTURE OF DICTYOCHA FIBULA EHRENBERG. II. THE PROTOPLAST1

Dictyocha fibula in exponential phase cultures displays a range of morphological variants of which the “sunburst” is most common. In this form, the perinuclear cytoplasm (perikaryon) contains an average of 72 dictyosomes, assorted vesicles, mitochondria, endoplasmic reticulum, and ribosomes. Cytoplasmic processes, globose to irregular, extend on fine cytoplasmic strands from the perikaryon into an extensive, viscous wall, structureless in electron micrographs except for scattered electron-opaque leaflets near the perikaryon. Mitochondria with tubular cristae occur within the globose process and occasionally within the connecting strands. Chloroplasts, with 3-disk lamellar bands and with pyrenoids not crossed by lamellae, are confined to the cytoplasmic processes in the sunburst from. A structure which may be the “flagellum” occasionally occurs attached to the perikaryon. However, no flagellar structures containing microtubules, nor flagellar root structures, have been found.     

FINE STRUCTURE AND REPLICATION OF THE KINETOPLAST OF TRYPANOSOMA LEWISI

The kinetoplastic DNA of Trypanosoma lewisi is described as a filamentous body lying within a mitochondrion, with the filaments oriented parallel to the long axis of the cell. The manner of fixation, the replicative state, and perhaps the physiological state of the cell, may result in slight morphological differences among such bodies. The kinetoplastic DNA replicates to form "left" and "right" rather than "upper" and "lower" members, and both the kinetoplast and nucleus incorporate radiothymidine as shown by radioautography. Radioautographic analyses suggest a random incorporation of radiothymidine by kinetoplasts. Silver grains were occasionally observed over centriolar elements. Finally, the observations are discussed with respect to the sequential replication of the aforementioned organelles by T. lewisi.