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.     

STRUCTURE AND DEVELOPMENT OF THE CHLOROPLAST IN CHLAMYDOMONAS : I. THE NORMAL GREEN CELL

The cytoplasmic organization of a normal green strain of the alga Chlamydomonas reinhardi has been investigated with the electron microscope using thin sections of OsO₄ fixed material. The detailed organization of the chloroplast has been of special interest. The chloroplast, a cup-shaped organelle, surrounded by a double membrane, consists of: (1) discs about 1 micron in diameter, considered to represent the basic structural unit of the chloroplast, and each composed of a pair of membranes joined at their ends to form a flat closed vesicle; the discs are grouped into stacks resembling the grana of higher plants; (2) matrix material of low density in which the discs are embedded; (3) starch grains; (4) the pyrenoid, a non-lamellar region associated with starch synthesis, and containing tubules which connect with the lamellae; (5) the eyespot, a differentiated region containing two or three plates of hexagonally packed, carotenoid-containing granules, located between discs, and associated with phototaxis. In addition to the chloroplast, the cytoplasm contains various membranous and granular components, including mitochondria, endoplasmic reticulum, and dictyosomes, identified on the basis of morphological comparability with structures seen in animal cells. The nucleus, not investigated in detail in this study, contains a large, granular nucleolus and is surrounded by a nuclear envelope which is provided with pores and exhibits instances of continuity with the endoplasmic reticulum of the cytoplasm.     

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.     

La Structure Fine de Chlorogonium elongatum. I. Etude Systematique au Microscope Electronique.

The flagellate Chlorogonium elongatum has been studied in the electron microscope by thin-sectioning techniques. The nucleus, limited by a porous membrane, contains a large nucleolus and some dense masses, probably of chromatin. When the medium is rich or the culture young, the chloroplast has few lamellae, and very few pyrenoids. Inversely, when the medium is poor or jthe culture old, the lamellae are abundant, appearing as piled discs or as a tortuous lamellar system. The pyrenoids are then numerous and are surrounded by starch grains. They are composed of an opaque, finely granular substance, and are never traversed by chloroplast lamellae. The stigma, within the chloroplast, is a circular or oval plate composed of a single tier of dense granulations. The cytoplasm contains mitochondria which are concentrated near the pellicle of the cell, an endoplasmic reticulum, an abundance of ribo-nucleoprotein particles or ribosomes, and a Golgi apparatus. These organelles resemble those observed in other protozoa. The two fiagella are encased in cylinders situated at the anterior end of the cell. At the base of the fiagella are the basal corpuscles or kinetosomes, which repose within a cupule. A dense granule which probably corresponds to the centrosome appears near this structure.     

ULTRASTRUCTURE OF THE LAMELLAE AND GRANA IN THE CHLOROPLASTS OF ZEA MAYS L

The fine structure of the chloroplasts of maize (Zea mays L.) has been investigated by electron microscopic examination of ultrathin sections of leaves fixed in buffered osmium tetroxide solutions. Both the parenchyma sheath and mesophyll chloroplasts contain a system of densely staining lamellae about 125 A thick immersed in a finely granular matrix material (the stroma), and are bounded by a thin limiting membrane which often appears as a double structure. In the parenchyma sheath chloroplasts, the lamellae usually extend the full width of the disc-shaped plastids, and grana are absent. The mesophyll chloroplasts, however, contain numerous grana of a fairly regular cylindrical form. These consist of highly ordered stacks of dense lamellae, the interlamellar spacing being ca. 125 A. The grana are interlinked by a system of lamellae (intergrana lamellae) which are on the average about one-half as numerous as the lamellae within the grana. In general, this appears to be due to a bifurcation of the lamellae at the periphery of the granum, but more complex interrelationships have been observed. The lamellae of the parenchyma sheath chloroplasts and those of both the grana and intergrana regions of the mesophyll chloroplasts exhibit a compound structure when oriented normally to the plane of the section. A central exceptionally dense line (ca. 35 A thick) designated the P zone is interposed between two less dense layers (the L zones, ca. 45 A thick), the outer borders of which are defined by thin dense lines (the C zones). Within the grana, the C zones, by virtue of their close apposition, give rise to thin dense intermediate lines (I zones) situated midway between adjacent P zones. A model of the lamellar structure is proposed in which mixed lipide layers (L zones) are linked to a protein layer (P zone) by non-polar interaction. Chlorophyll is distributed over the entire lamellar surface and held in the structure by van der Waals interaction of the phytol "tail" with the hydrocarbon moieties of the mixed lipide layers. The evidence in favour of the model is briefly discussed.     

ELECTRON MICROSCOPY OF THE VOLVOCACEAE AND ASTREPHOMENACEAE

Lang, Norma J. (U. Texas, Austin.) Electron microscopy of the Volvocaceae and Astrephomenaceae. Amer. Jour. Bot. 50(3): 280-300. Illus. 1963.—Clonal cultures of Gonium sociale, G. pectorale, Pandorina morum, Eudorina elegans, Eudorina sp., Volvulina steinii, V. pringsheimii, Platydorina caudata, Pleodorina illinoisensis, P. californica, Volvox aureus, V. tertius, V. globator, V. barberi, and Astrephomene gubernaculifera representing the Volvocaceae and Astrephomenaceae in the Volvocales were examined with the electron microscope and their ultrastructure compared. The ultrastructure of the various organelles is basically similar in the species studied and no increase in cellular complexity is found to accompany the evolutionary trends evidenced in the Volvocaceae. The ultrastructure of a colonial cell is basically that of Chlamydotnonas. A cytoplasmic membrane having a unit membrane structure encompasses a cell and is continuous with the double-membraned flagellar sheaths. The flagella contain the typical 9 + 2 fibril arrangement with the 2 axial fibrils terminating in a cylinder at the flagellar base and the 9 peripheral pairs continuing into the cytoplasm as a basal body. The organelles comprising the cytoplasm are: mitochondria with plate-like cristae; dictyosomes composed of stacks of agranular cisternae; small, rough or smooth-surfaced vesicles; an endoplasmic reticulum of granule-bearing and agranular tubules, lamellae and broad cisternae; vacuoles which are either contractile, contain fine granular and fibrillar material, or have dense contents probably representing polyphosphate; lipid bodies; and dense granules 100–150 A which have been called ribosomes. The finely granular nucleoplasm is surrounded by a porous, double-membraned nuclear envelope and contains a centric nucleolus composed of dense, spherical granules. The outer membrane of the nuclear envelope bears granules and may have granular extensions into the perinuclear cytoplasm. Each extension appears to encompass one or several dictyosomes and has been termed an “amplexus.” The amplexi are agranular on the surface contiguous to a dictyosome. A double-membraned chloroplast envelope is continuous around the single, cup-shaped chloroplast. The basic chloroplast units are discs closed at each end, occurring in stacks of varying number parallel to the envelope. The presumed proteinaceous matrix of the basal pyrenoid is penetrated by elongated, tubular elements which connect with the lamellar discs. Multiple rows of granules, associated with individual discs, form the anterior stigma within the chloroplast envelope. The colonial matrix is not a structureless, mucilaginous material uniting the cells in colonies, but it has rather a highly complex structure especially around the periphery of the colony and the flagellar channels. The apparent substitution of a fibrillar layer of the colonial matrix for the discrete compact cell wall, such as is found in Chlamydomonas, implies a greater degree of complexity in the evolution of these colonial genera than is generally assumed.     

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.     

OOCYTE DIFFERENTIATION IN THE SEA URCHIN, ARBACIA PUNCTULATA, WITH PARTICULAR REFERENCE TO THE ORIGIN OF CORTICAL GRANULES AND THEIR PARTICIPATION IN THE CORTICAL REACTION

This paper presents morphological evidence on the origin of cortical granules in the oocytes of Arbacia punctulata and other echinoderms. During oocyte differentiation, those Golgi complexes associated with the production of cortical granules are composed of numerous saccules with companion vesicles. Each element of the Golgi complex contains a rather dense homogeneous substance. The vesicular component of the Golgi complex is thought to be derived from the saccular member by a pinching-off process. The pinched-off vesicles are viewed as containers of the precursor(s) of the cortical granules. In time, they coalesce and form a mature cortical granule whose content is bounded by a unit membrane. Thus, it is asserted that the Golgi complex is involved in both the synthesis and concentration of precursors utilized in the construction of the cortical granule. Immediately after the egg is activated by the sperm the primary envelope becomes detached from the oolemma, thereby forming what we have called the activation calyx (see Discussion). Subsequent to the elaboration of the activation calyx, the contents of cortical granules are released (cortical reaction) into the perivitelline space. The discharge of the constituents of a cortical granule is accomplished by the union of its encompassing unit membrane, in several places, with the oolemma.     

THE FINE STRUCTURE OF THE NUCLEOLUS DURING MITOSIS IN THE GRASSHOPPER NEUROBLAST CELL

The behavior of the nucleolus during mitosis was studied by electron microscopy in neuroblast cells of the grasshopper embryo, Chortophaga viridifasciata. Living neuroblast cells were observed in the light microscope, and their mitotic stages were identified and recorded. The cells were fixed and embedded; alternate thick and thin sections were made for light and electron microscopy. The interphase nucleolus consists of two fine structural components arranged in separate zones. Concentrations of 150 A granules form a dense peripheral zone, while the central regions are composed of a homogeneous background substance. Observations show that nucleolar dissolution in prophase occurs in two steps with a preliminary loss of the background substance followed by a dispersal of the granules. Nucleolar material reappears at anaphase as small clumps or layers at the chromosome surfaces. These later form into definite bodies, which disappear as the nucleolus grows in telophase. Evidence suggests both a collecting and a synthesizing role for the nucleolus-associated chromatin. The final, mature nucleolar form is produced by a rearrangement of the fine structural components and an increase in their mass.     

ELECTRON MICROSCOPY OF THE PACINIAN CORPUSCLE

The Pacinian corpuscle has a framework of cytoplasmic lamellae arranged concentrically in the outer zone, and bilaterally in the core. Between these is an intermediate growth zone. The inner core shows an unexpected complexity in that its component lamellae are arranged in two symmetrical groups of nested cytoplasmic sheets. Longitudinal tissue spaces form clefts separating the two groups. The perikarya of the core lamellae lie in or near the intermediate growth zone, and send arms into the clefts. The arms then branch and terminate as lamellae which interdigitate with those of neighboring cells. The single nerve fiber loses its myelin sheath just before it reaches the inner core but retains its Schwann cell cytoplasmic covering for a short additional distance. The Schwann sheath is not continuous with the lamellae of the inner core. Inside the core the fiber contains a striking circumferential palisade of radially disposed mitochondria. The fiber does not arborize. Vascular capillaries penetrate the hilar region of the corpuscle only as far as the myelinated sheath of the nerve, and they have not been seen elsewhere in the corpuscle. There is direct continuity between the clefts of the core and tissue spaces in the vicinity of the capillaries. It is likely that this provides a route whereby metabolites reach the active nerve ending, as well as the cells of the growth zone. The outer zone consists of at least 30 flattened concentric cytoplasmic lamellae separated from one another by relatively wide fluid-filled spaces. Collagenous fibrils are present, particularly on the outer surface of lamellae, and tend to be oriented circularly. The girdle of proliferating cells constituting the growth zone, which is prominent in corpuscles from young animals, is the layer from which the outer lamellae are derived. Osmotic forces probably elevate the lamellae, and maintain turgor pressure.     

THE ISOLATION OF A MAJOR STRUCTURAL ELEMENT OF THE SEA URCHIN FERTILIZATION MEMBRANE

Procedures for isolating the contents of the cortical granules from the ova of the sea urchin, Strongylocentrotus purpuratus, are reported. Dithiothreitol is used to remove the vitelline coat; the "demembranated" eggs are then subsequently activated with butyric acid. By means of these procedures, the hyaline protein and crystalline or paracrystalline material have been isolated from the cortical granules. The crystalline material consists of sheets of cylinders or tubules 150–200 A in diameter. This material is believed to be a major structural element of the fertilization membrane which, in the absence of the vitelline coat, does not form.     

Presence of acid phosphatase activity in heavy bodies of sea urchin eggs

Cytochemical studies were made on the acid phosphatase (Acp-ase) in the heavy bodies of eggs of the sea urchins, Hemicentrotus pulcherrimus and Temnopleurus toreumaticus, from the stage of mature unfertilized eggs to the late gastrula stage. High Acp-ase activity was found in masses composed of ribosome-like granules in the heavy bodies from the early stage of the formation of heavy bodies until the time of their disappearance, whereas the annulate lamellae and the space between the annulate lamellae and the masses packed with ribosome-like granules did not have activity. This activity was inhibited by the addition of 0.01 M NaF to, or the omission of substrate from, the Gomori medium. Using the reaction products of Acp-ase and the masses of ribosome-like granules as markers of heavy bodies, we found small groups of tightly packed ribosome-like granules that were not surrounded by annulate lamellae. The function of Acp-ase in the heavy bodies is still unknown.     

FINE STRUCTURE AND MORPHOGENIC MOVEMENTS IN THE GASTRULA OF THE TREEFROG, HYLA REGILLA

The blastoporal groove of the early gastrula of the treefrog, Hyla regilla, was examined with the electron microscope. The innermost extension of the groove is lined with invaginating flask- and wedge-shaped cells of entoderm and mesoderm. The distal surfaces of these cells bear microvilli which are underlain with an electron-opaque layer composed of fine granular material and fibrils. The dense layer and masses of vesicles proximal to it fill the necks of the cells. In flask cells bordering the forming archenteron the vesicles are replaced by large vacuoles surrounded by layers of membranes. The cells lining the groove are tightly joined at their distal ends in the region of the dense layer. Proximally, the cell bodies are separated by wide intercellular spaces. The cell body, which is migrating toward the interior of the gastrula, contains the nucleus plus other organalles and inclusions common to amphibian gastrular cells. A dense layer of granular material, vesicles, and membranes lies beneath the surface of the cell body and extends into pseudopodium-like processes and surface undulations which cross the intercellular spaces. A special mesodermal cell observed in the dorsal lining of the groove is smaller and denser than the surrounding presumptive chordamesodermal cells. A long finger of cytoplasm, filled with a dense layer, vesicles and membranes, extends from its distal surface along the edge of the groove, ending in a tight interlocking with another mesodermal cell. Some correlations between fine structure and the mechanics of gastrulation are discussed, and a theory of invagination is proposed, based on contraction and expansion of the dense layer and the tight junctions at distal cell surfaces.     

A POSSIBLE MECHANISM FOR THE MORPHOGENESIS OF LAMELLAR SYSTEMS IN PLANT CELLS

A mechanism for the formation of lamellar systems in the plant cell has been proposed as a result of electron microscope observations of young and mature cells of Nitella cristata and the plastids of Zea mays in normal plants, developing plants, and certain mutant types. The results are compatible with the concept that lamellar structures arise by the fusion or coalescence of small vesicular elements, giving rise initially to closed double membrane Structures (cisternae). In the chloroplasts of Zea, the cisternae subsequently undergo structural transformations to give rise to a compound layer structure already described for the individual chloroplast lamellae. During normal development, the minute vesicles in the young chloroplast are aggregated into one or more dense granular bodies (prolamellar bodies) which often appear crystalline. Lamellae grow out from these bodies. In fully etiolated leaves lamellae are absent and the prolamellar bodies become quite large, presumably because of inhibition of the fusion step which appears to require chlorophyll. Lamellae develop rapidly on exposure of the plant to light, and subsequent development closely parallels that seen under normal conditions. The plastids of white and very pale green mutants of Zea similarly lack lamellae and contain only vesicular elements. A specialized peripheral zone immediately below the double limiting membrane in Zea chloroplasts appears to be responsible for the production of vesicles. These may be immediately converted to lamellae under normal conditions, but accumulate to form a prolamellar body if lamellar formation is prevented, as in the case of etiolation and chlorophyll-deficient mutation, or when the rate of lamellar formation is slower than that of the production of precursor material (as appears to be the case in the early stages of normal development).     

THE GOLGI APPARATUS IN NEURONS AND EPITHELIAL CELLS OF THE COMMON LIMPET PATELLA VULGATA

1. In view of widely diverse views held about the identity and structure of the Golgi apparatus in neurons of Mollusca, particularly gastropods, a study has been made on neurons of the common limpet, Patella vulgata, both by light and electron microscopy. A report is given also of observations made on epithelial cells of Patella by electron microscopy. 2. As revealed by Kolatchev's method, the Golgi apparatus in neurons consists basically of black filaments lying to one side of the nucleus. The filaments generally anastomose to form networks of various complexity. Rarely some cells contain only discrete filaments. Associated with some of the filaments is a weakly osmiophilic substance identified as archoplasm. Kolatchev's method also revealed spheroidal bodies (neutral red bodies, "lipochondria," etc.). 3. It has not been possible to demonstrate the Golgi apparatus using either iron-haematoxylin or Sudan black. 4. Examination of Kolatchev's preparations by electron microscopy has revealed that some of the Golgi filaments consist of chromophilic and chromophobic components. The chromophilic component consists of dense lamellae. 5. After fixation in buffered osmium tetroxide solution and examination by electron microscopy, it has been concluded that (a) the chromophilic component of the Golgi apparatus corresponds to a system of paired membranes (which usually enclose an inner dense substance), (b) the chromophobic component corresponds to a substance lying within small dilations of the paired membrane, and (c) the archoplasm corresponds to numerous small vesicles. 6. The paired membranes branch, anastomose, and can often be traced back to a common source. They are interpreted as lamelliform folds, and occasionally tubular processes, of essentially a single Golgi membrane. In cells containing a Golgi network it is suggested that the membrane extends through the whole of the apparatus in such a way that the substance it encloses may be regarded as being in a continuous phase. 7. Epithelial cells of Patella contain a juxtanuclear Golgi apparatus with an ultrastructure similar to that described for neurons.     

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 mmicro 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.     

豚鼠Clara细胞GERL及其在低氧下变化的细胞化学观察

本文对正常和模拟海拔5000米高度24、48和72小时后豚鼠Clara细胞GERL进行了超微结构和ACP活性的电镜酶细胞化学观察。结果表明Clara细胞具发达的GERL,包括ACP阳性的致密膜囊、致密囊泡、有衣小泡和细胞衣被覆区,Clara细胞GERL参与了分泌颗粒的形成。在低氧下GERL肥大,ACP阳性分泌颗粒增多,提示低氧导致GERL功能亢进,形成分泌颗粒的作用增强。     

The histochemistry and ultrastructure of the clitellum of the enchytraeid Lumbricittus rivalis (Oligochaeta: Annelida)

The monostratified clitellar epidermis of Lumbricillus rivalis consists of supporting cells, granular secretory cells, and globular secretory cells, together with the acid mucous cells typical of normal skin in the anterior and posterior transition zones. The secretion of the granular cells is a neutral glycoprotein with low levels of bound lipid, and that of the globular cells is a sulphated acid mucopolysaccharide lacking detectable protein or lipid.
Ultrastructurally the granular cells possess 1 μm, membrane-bound granules of variable electron density suggestive of maturation changes. The formative granules arise from Golgi vesicles and are moderately electron translucent. They contain parallel-aligned, tubule-like inclusions of 14 nm diameter, observed also in the mature regions of the Golgi. Granules in the mid or apical part of the cell show increased electron density, the formative pattern being wholly or partially obscured and the subunit alignment, when discernable, no longer uniform. Moiré fringe patterns are evident in some granules. The patterns and electron density are lost after pronase digestion. The globular cells contain electron translucent, membrane-bound globules of 2–3 μm diameter with finely fibrous contents. They arise from mature Golgi vesicles and are unaffected by pronase treatment.
The results suggest that the granular cells secrete the cocoon wall and the globular cells the material that surrounds the developing embryos, and are compared with published accounts of other microdrile clitella and with those of the clitellum of lumbricid earthworms.