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.     

A quantitative analysis of the incidence of certain cytoplasmic structures in the ovum of the rat during cleavage

Summary The volumetric density of most cytoplasmic organelles in the segmenting ovum of the rat was determined by morphometric techniques described by Weibel, at the 1-, 2-, 4-, and 8-cell stages, and at the early and late blastocyst stages.During the course of cleavage, the volumetric density of mitochondria remained practically unchanged. The volumetric density of the Golgi complex was too small to permit assessment of the differences between the individual stages of development. Significant changes were found in both granular and agranular endoplasmic reticulum, secondary lysosomes, multivesicular bodies, lamellar structures and lipid droplets. Granular endoplasmic reticulum was first observed as late as the 8-cell ovum stage and its volumetric density increased further in the early and late blastocysts. The relative volume of agranular endoplasmic reticulum was highest in the 1-cell ovum and decreased during the course of cleavage. The same is true for multivesicular bodies. The volumetric density of secondary lysosomes increased during cleavage, reaching the highest values in the 8-cell ovum. Lamellar structures were the most voluminous part of the cytoplasm of the segmenting ovum at all stages. Their volumetric density, however, decreased during the course of cleavage. Lipid droplets occur in very small quantities in the 1- to 4-cell ova, but at later stages their volumetric density increased.Our findings underline the importance of acquiring quantitative information about changes in cell organelle populations for assessing morphological and functional relationships during the early stages of cleavage of the ovum.     

Fine structure of spermatogonia and primary spermatocytes in rabbits

Summary The fine structure of rabbit Spermatogonia and primary spermatocytes in meiotic prophase has been studied with different methods of preparation, including a technique for acid phosphatase activity. The spermatogonial cytoplasm is rich in free ribosomes and containes moderate amounts of vesicular, smooth-surfaced endoplasmic reticulum and mitochondria, a simple Golgi-apparatus, some micropinocytotic vesicles, and occasional multivesicular bodies, vacuoles and dense bodies with acid phosphatase activity. The large type A Spermatogonia have a prominent nucleolus and their mitochondria sometimes form clusters with a dense intermitochondrial substance, similar to that in spermatocytes.The nucleus and cytoplasm of primary spermatocytes increase markedly in volume and density during meiotic prophase. The Golgi apparatus enlarges and becomes more differentiated and finally forms small proacrosome granules. The endoplasmic reticulum produces numerous small, mainly smooth vesicles and might also be the source of a new organelle: numerous piles of narrow cisternae with opaque contents. These piles disintegrate late in prophase. The mitochondria become aggregated in clusters with dense intermitochondrial substance and their internal structure is characterized by highly dilated cristae and small particles, interpreted as mitochondrial ribosomes, in the matrix. The role of these structures in the formation of new mitochondria is discussed. The clusters of mitochondria finally disperse and their cores of dense intermitochondrial substance, possibly containing ribonucleoprotein, coalesce into a large chromatoid body similar to that in spermatids. Micropinocytosis and a few lysosomes occur in most spermatocytes. The pachytene nuclei show prominent nucleoli and a distinct sex vesicle without any synaptinemal complex.The importance for spermatid differentiation of some events taking place in the cytoplasm of primary spermatocytes is emphasized.Financial support for this study was received from the Swedish Medical Research Council.     

Light and electron microscopic immunocytochemical localization of clathrin in rat cerebellum and kidney

The precise cellular and subcellular locations of coated vesicle protein, clathrin, in rat kidney and cerebellum have been visualized by immunocytochemical techniques. In the renal tubular epithelia, clathrin-positive products were found on both free ribosomes and on those attached to rough endoplasmic reticulum (RER) and the nuclear envelope. No clathrin was observed in the cisternae of RER or the Golgi apparatus. Clathrin-positive reaction products could also be seen on coated pits, coated vesicles, Golgi-associated vesicles, basolateral cell membrane, the ground substance, and in the autophagic vacuoles. In cerebellar Purkinje and granule cell bodies, reaction products were seen localized on coated vesicles, on the budding areas from the Golgi-associated membrane and Golgi-associated vesicles. Furthermore, the membrane of the multivesicular body, the bound-ribosomes, and the ground substance were also stained. In the myelinated axon, the clathrin appeared to be concentrated on certain segments and seemed to fill in the space between neurotubules and some vesicles. In certain synaptic terminals clathrin was often seen attached to presynaptic vesicles, presynaptic membrane, and post-synaptic membrane. However, in most mossy fibers, some synaptic vesicles were not stained. These observations suggest that clathrin is synthesized on bound and free ribosomes and discharged into the cytosol where it becomes associated with a variety of ground substances and assembles on coated pits, coated vesicles, Golgi-associated vesicles, presynaptic vesicles, and pre- and postsynaptic membranes. Clathrin may be finally degraded in autophagic vacuoles.     

Twenty-four-hour variations in subcellular structures of rat type II alveolar epithelial cells

Summary Subcellular structures of type II alveolar epithelial cells in the rat lung were analyzed at six evenly spaced times over 24 h (light period: 06.00 h–18.00 h), using a morphometric technique. The cell volumes were maximal at 16.00 h and minimal at 08.00 h. The volume and surface densities of rough endoplasmic reticulum and mitochondria were low during the light period, and high during the dark period. Morphometric parameters of multivesicular bodies did not significantly fluctuate over 24 h, but they increased from 04.00 h to 08.00 h. The volume densities of lamellar bodies increased from 16.00 h to 20.00 h, and decreased from 00.00 h to 08.00 h. The change in numerical densities of lamellar bodies was inversely correlated to that in the volume densities. As shown by electron microscopy, small lamellar bodies predominated at 08.00 h, larger lamellar bodies increasing at 16.00h. Composite bodies often appeared at 08.00 h and 12.00 h. Type II cells thus appear to fluctuate, showing three phases over 24 h: formation, accumulation and secretion of lamellar bodies. In particular, it is noteworthy that the accumulation stage occurs during the resting phase of the rat, whereas the secretion stage occurs during its body-active phase.     

Genital primordium of the free-living marine nematode Halichoanolaimus sonorus (Chromadoria,Chromadorida)

Summary

The genital primordium of the first stage juvenile (J1) of the free-living marine nematode Halichoanolaimus sonorus (Chromadorida: Selachinematidae) was studied using transmission electron microscopy. The primordium consists of four undifferentiated cells: two primordial germ cells (PGC) 5–6 μm in diameter and two somatic cells. The PGC have a large nucleus with nucleolus. The centriole was detected in close vicinity of the PGC nucleus. Most of the cell mitochondria are in close contact with the nuclear envelope. The mitochondria are interspersed by 0.2–0.3 μm particles of an electron-dense diffuse substance devoid of surrounding membrane. Both PGC are closely attached to each other and to the neighboring somatic cells of the genital primordium. The elongated somatic cells contain nuclei devoid of nucleoli; the cytoplasm is filled with free ribosomes and contains occasional cisternae of rough endoplasmatic reticulum (RER), Golgi bodies, mitochondria, and transparent vesicles. The genital primordium is separated by a narrow space from of the intestine (dorsally) and the somatic muscles (ventrally). The PGC of H. sonorous are devoid of typical P granules known for previously studied nematodes as distinct markers of germ line cell lineage. Perinuclear particles of dense diffuse substance found in PGC of H. sonorous could be considered as germ determinants analogous to P granules.     

PATTERN OF OSMIUM DEPOSITION IN THE PARIETAL CELLS OF THE STOMACH

Parietal cells of the stomach of the hamster show extensive amounts of dense material in a variety of organelles after prolonged exposure to a solution of osmium tetroxide. Conspicuous amounts of reduced osmium compounds are evident within the granular endoplasmic reticulum, perinuclear cisterna, and vesicular elements of the Golgi complex. Dense material is also apparent within cristae of the mitochondria, the surface coat of the microvilli of the intracellular canaliculus, and vesicular elements of the multivesicular bodies. Multivesicular bodies, containing numerous small osmiophilic elements, are often seen surrounding and/or in close contact with mitochondria. The proximity of the multivesicular bodies to the mitochondria appears to be related to an autophagic process involving degradation of mitochondria. The distribution and intensity of the precipitates within the organelles of the parietal cells vary in different regions of the gastric glands. The findings of this study emphasize that cell structures other than the Golgi complex may contain large concentrations of reduced osmium compounds after prolonged exposure to a solution of osmium tetroxide.     

The Ultrastructure of Lankesterella hylae

SYNOPSIS. The ultrastructure of Lankesterella hylae was studied and numerous points of similarity to Plasmodium, Toxoplasma, Sarcocystis and Lankesterella garnhami were found. The protozoa were intracellular and lay within vacuoles containing vesicles, unusual membrane formations and dense granular material. The parasite was invested by a double membrane and had a micropyle, as well as membranous processes extending from the surface. At the anterior end were conoid and apical rings. The cell contained a nucleus, nucleolus, bipolar paranuclear vacuoles or bodies, a series of microtubules beneath the pellicle, endoplasmic reticulum, mitochondria, toxonemes and a variety of vacuoles. In addition, dense particles, similar to those related to the endoplasmic reticulum, were scattered throughout the cytoplasm.
The unusual membrane formations and vesicles in the periparasitic vacuoles were striking observations possibly related to the nutrition of the parasite.     

Effects of starvation on the ultrastructure of the mouse parathyroid gland

Ultrastructural changes of the parathyroid glands of starved mice were examined. The parathyroid glands of the starved mice showed a decrease in the volume of Golgi complexes and storage granules and an increase in the volume of lipid droplets, and contained more heterogeneously dense bodies and multivesicular bodies compared with that of the control mice. In addition, the volume of mitochondria, cisternae of granular endoplasmic reticulum and secretory granules and the number of prosecretory granules appeared to be decreased compared to those of the control mice. Myelin-like structures were observed in the parathyroid glands of the starved mice. The results of our study provide support for the hypothesis that starvation exerts an inhibitory influence not only on the synthesis but also on release of parathyroid hormone.     

薏苡种子胚芽鞘细胞的结构

观察了薏苡浸泡种子胚芽鞘的结构。胚芽 外,内表皮薄壁组织及２个侧位的维管束组成。在外表皮两处,观察到径向壁不边原细胞群,它们实际是合胞体。薄壁细胞含丰富的核糖体,内质网小泡和线粒体,说明代谢活动已经活跃。初生纹孔场内有胞间连丝,显示胞间已存在物质的共质运转。     

Stereological analysis of hepatic fine structure in the Fischer 344 rat. Influence of sublobular location and animal age

Stereological analysis of hepatic fine structure in Fischer 344 male rats at 1, 6, 10, 16, 20, 25, and 30 mo of age revealed differences in the amounts and distributions of hepatocellular organelles as a function of sublobular location or animal age. Between 1 and 16 mo of age, both the centrolobular and periportal hepatocytes increased in volume by 65 and 35%, respectively. Subsequently, the cell volumes declined until the hepatocytes of 30-mo-old rats approached the size of those found in the youngest animals. Regardless of animal age, the centrolobular cells were consistently larger than the corresponding periportal hepatocytes. The cytoplasmic and ground substance compartments reflected similar changes in their volumes, although there was no significant alteration in the nuclear volume. The volumes of the mitochondrial and microbody compartments increased and decreased concomitant with the changes in average hepatocyte size. Both lobular zones in the 30-mo-old rats contained significantly smaller relative volumes of mitochondria than similar parenchyma in 16-mo-old animals. The volume density of the dense bodies (lysosomes) increased markedly in both lobular zones between 1 and 30 mo of age, confirming reports of an age-dependent increase in this organelle. The surface area of the endoplasmic reticulum in the centrolobular and periportal hepatocytes reached its maximum level in the 10-mo-old rats and subsequently declined to amounts which approximated those measured in the 1-mo-old animals. This age-related loss of intracellular membrane is attributable to a significant reduction in the surface area of the smooth-surfaced endoplasmic reticulum (SER) in animals beyond 16 mo of age. The amount of rough-surfaced endoplasmic reticulum (RER) in the periportal parenchymal cells was unaffected by aging, but the centrolobular hepatocytes of 30-mo-old animals contained 90% more RER than similar cells in the youngest rats. The centrolobular parenchyma contained more SER and the portal zones more RER throughout the age span studied. These quantitative data suggest that (a) certain hepatic fine structural parameters undergo marked changes as a function of animal age, (b) there exists a gradient in hepatocellular fine structure across the entire liver lobule, and (c) there are remarkable similarities in hepatocyte ultrastructure between very young and senescent animals, including cell size and the amount of SER.     

The inositol 1,4,5,-trisphosphate receptor in cerebellar Purkinje cells: quantitative immunogold labeling reveals concentration in an ER subcompartment

The Ca2+ mobilization effect of inositol 1,4,5-trisphosphate, the second messenger generated via receptor-stimulated hydrolysis of phosphatidylinositol 4,5-bisphosphate, is mediated by binding to intracellular receptors, which are expressed in high concentration in cerebellar Purkinje cells. Partially conflicting previous reports localized the receptor to various subcellular structures: elements of ER, both rough and smooth-surfaced, the nuclear envelope, and even the plasma membrane. We have now reinvestigated the problem quantitatively by using cryosections of rat cerebellar tissue immunolabeled with polyclonal monospecific antibodies against the inositol 1,4,5-trisphosphate receptor. By immunofluorescence the receptor was detected only in Purkinje cells, whereas the other cells of the cerebellar cortex remained negative. In immunogold-decorated ultrathin cryosections of the Purkinje cell body, the receptor was concentrated in cisternal stacks (piles of up to 12 parallel cisternae separated by regularly spaced bridges, located both in the deep cytoplasm and beneath the plasma membrane; average density, greater than 5 particles/micron of membrane profile); in cisternal singlets and doublets adjacent to the plasma membrane (average density, approximately 2.5 particles/micron); and in other apparently smooth-surfaced vesicular and tubular profiles. Additional smooth-surfaced elements were unlabeled. Perinuclear and rough-surfaced ER cisternae were labeled much less by themselves (approximately 0.5 particles/micron, two- to threefold the background), but were often in direct membrane continuity with heavily labeled, smooth-surfaced tubules and cisternal stacks. Finally, mitochondria, Golgi cisternae, multivesicular bodies, and the plasma membrane were unlabeled. In dendrites, approximately half of the nonmitochondrial, membrane-bound structures (cisternae, tubules, and vesicles), as well as small cisternal stacks, were labeled. Dendritic spines always contained immunolabeled cisternae and vesicles. The dendritic plasma membrane, of both shaft and spines, was consistently unlabeled. These results identify a large, smooth-surfaced ER subcompartment that appears equipped to play a key role in the control of Ca2+ homeostasis: in particular, in the generation of [Ca2+]i transients triggered by activation of specific receptors, such as the quisqualate-preferring trans(+/-)-1-amino-1,3-cyclopentamedicarboxylic acid glutamatergic receptors, which are largely expressed by Purkinje cells.     

Ultrastructural distribution of Ca++ within neurons. An oxalate pyroantimonate study

We used the oxalate-pyroantimonate technique to determine the ultrastructural distribution of Ca++ in neurons of the rat sciatic nerve. The content of the precipitate was confirmed by X-ray microanalysis and appropriate controls. In the cell bodies of the dorsal root ganglia, Ca++ precipitate was found in the Golgi, mitochondria, multivesicular bodies and large vesicles of the cytoplasm but not in lysosomes, and was prominently absent from regions of rough endoplasmic reticulum and ribosomes. It was seen in the nucleus but not in the nuclear bodies or nucleolus. Within the axon itself, Ca++ precipitate was also found sequestered in mitochondria and smooth endoplasmic reticulum. In addition Ca++ precipitate found diffusely throughout the axoplasm exhibited a discrete and heterogeneous distribution. In myelinated fibers the amount of precipitate decreased predictably in the axoplasm beneath the Schmidt-Lanterman cleft and in the paranodal regions at the nodes of Ranvier. This correlated with the presence of dense precipitate in the Schmidt-Lanterman cleft themselves and in the paranodal loops of myelin. Intracytoplasmic ionic Ca++ is maintained at 10(-7) M by balanced processes of influx, sequestration and extrusion. The irregular distribution of Ca++ precipitate in the axoplasm of myelinated fibers suggests that there may be specific regions of preferential efflux across the axolemma.     

Morphometric study of normal and phytohemagglutinin-stimulated lymphocytes

Summary The morphological changes of human lymphocytes living in vitro under the influence of phytohemagglutinin (PHA) were estimated quantitatively by stereological methods. Mean volume and volume fractions for average human lymphocyte and average blastoid cell from 72-hour culture with PHA were determined. It was found that the lymphocyte volume increases on an average 3.7 times mainly because of cytoplasmic volume increment by 4.3 times. The greatest enlargement concerned endoplasmic reticulum, the nucleolus, lysosomes and mitochondria. In addition a way of correction of the error in experimentally obtained volume fractions, resulting from the exclusive use of sections involving the nucleus is presented.     

Alterations Induced by the Antifungal Compounds Ketoconazole and Terbinafine in Leishmania

ABSTRACT. The antiproliferative effects and ultrastructural alterations induced in vitro by two antifungal compounds, the azole ketoconazole and the allylamine terbinafine on Leishmania amazonensis are reported. Promastigotes treatment with ketoconazole and terbinafine induced growth arrest and cell lysis in 72 hours. Combination of the two agents produced additive effects on promastigote axenic growth and synergistic effects on intracellular amastigote proliferation. The amastigotes, either axenically grown or infecting murine macrophages, were about 100-fold more sensitive to the drugs. These compounds induced the appearance of large multivesicular bodies, especially after ketoconazole treatment, increased amount of lipid inclusions as well as numerous, polymorphic volutin granules, particularly in terbinafine-treated cells. Multivesicular bodies were observed in close apposition with organelles such as mitochondria, which also showed alterations in the distribution and appearance of cristae, and the formation of paracrystalline arrays within the matrix. Some cells presented large portions of cytoplasm wrapped by endoplasmic reticulum and many parasites also presented myelin-like endoplasmic reticulum profiles. Such alterations together with the strong acid phosphatase activity observed in the multivesicular bodies and volutin granules may indicate the existence of an unusual autophagic process in cells treated with ergosterol biosynthesis inhibitors.     

Effect of tobacco mosaic virus strains on the ultrastructure of tobacco leaf parenchymal cells

Accumulation of considerable amounts of viral particles has been demonstrated in parenchymal cells of young leaves in tobacco cultivar Samsun systemically infected with any of studied tobacco mosaic virus (TMV) strains isolated from pepper (TMV-p), tomato (TMV-t), and eggplant (TMV-e). Abnormal (swollen and thin) virions were found, which points to their destruction. Cell infection with all studied strains was accompanied by the activation of the lysosomal compartment manifested as formation of nascent dictyosomes, elements of smooth endoplasmic reticulum, cytoplasmic vacuoles, various vesicles, invaginated mitochondria, and multivesicular bodies. The studied viral strains could be arranged in the following sequence according to the degree of lysosomal compartment stimulation and induction of intracellular lytic processes mediating the destruction of viral particles and cell structures: TMV-p > TMV-e > TMV-t.     

Immuno-gold localisation of arabinogalactan protein in the developing style of Nicotiana alata

Arabinogalactan-protein was localised ultrastructurally in the transmitting tissue of the Nicotinana alata Link & Otto style from the bud stage to anthesis using monoclonal antibody directed to terminal α-L-arabinosyl residues followed by goat anti-mouse IgG linked to 20 nm gold particles. Gold particles were localised on the Golgi apparatus, multivesicular bodies, cell wall and intercellular matrix with the highest concentration over multivesicular bodies. During flower development there was an increase in the total number of gold particles with a relative decrease on the multivesicular bodies and an increase on the intercellular matrix. There was also an increase during development in the proportion of intercellular matrix relative to all other components of the transmitting tissue. Vesiculation of the endoplasmic reticulum and the production of cup-shaped vesicles by the Golgi bodies were observed during development. It is concluded that secretion of arabinogalactan-protein from the intracellular to the extracellular location occurs via multivesicular bodies produced by the Golgi apparatus and possibly the endoplasmic reticulum.     

LYSOSOMES AND GERL IN NORMAL AND CHROMATOLYTIC NEURONS OF THE RAT GANGLION NODOSUM

The rat ganglion nodosum was used to study chromatolysis following axon section. After fixation by aldehyde perfusion, frozen sections were incubated for enzyme activities used as markers for cytoplasmic organelles as follows: acid phosphatase for lysosomes and GERL (a Golgi-related region of smooth endoplasmic reticulum from which lysosomes appear to develop) (31–33); inosine diphosphatase for endoplasmic reticulum and Golgi apparatus; thiamine pyrophosphatase for Golgi apparatus; acetycholinesterase for Nissl substance (endoplasmic reticulum); NADH-tetra-Nitro BT reductase for mitochondria. All but the mitochondrial enzyme were studied by electron microscopy as well as light microscopy. In chromatolytic perikarya there occur disruption of the rough endoplasmic reticulum in the center of the cell and segregation of the remainder to the cell periphery. Golgi apparatus, GERL, mitochondria and lysosomes accumulate in the central region of the cell. GERL is prominent in both normal and operated perikarya. Electron microscopic images suggest that its smooth endoplasmic reticulum produces a variety of lysosomes in several ways: (a) coated vesicles that separate from the reticulum; (b) dense bodies that arise from focal areas dilated with granular or membranous material; (c) "multivesicular bodies" in which vesicles and other material are sequestered; (d) autophagic vacuoles containing endoplasmic reticulum and ribosomes, presumably derived from the Nissl material, and mitochondria. The number of autophagic vacuoles increases following operation.     

Shedding of peripheral cytoplasm - a mechanism of liver cell atrophy in human amyloidosis.

A liver biopsy specimen from a case of primary amyloidosis was investigated by electron microscopy. The cytoplasmic periphery of the hepatocytes showed degenerativechanges which are interpreted as indicating shedding of peripheral parts of the cytoplasm. Two main variants of this process could be discerned: 1) Protrusion and sequestration of hernia-like blebs of cytoplasm, and 2) shedding of vesicles derived from degenerated endoplasmic reticulum. In the latter case transient defects of the plasma membrane seem to be relevance. Endoplasmic reticulum and cytoplasmic ground substance appeared to be shed preferentially, whereas mitochondria are retained within the cell. As a consequence the fractional volume of the mitochondria in the cytoplasm of atrophic cells is markedly increased. Shedding of peripheral cytoplasm, therefore, seems to be an effective mechanism enabeling the cell to adapt the mass and the composition of its cytoplasm to an unfavourable environment.     

Localization of calmodulin in rat cerebellum by immunoelectron microscopy

Calmodulin, a multifunctional Ca(++)-binding protein, is present in all eucaryotic cells. We have investigated the distribution of this protein in the rat cerebellum by immunoelectron microscopy using a Fab-peroxidase conjugate technique. In Purkinje and granular cell bodies, calmodulin reaction product was found localized both on free ribosomes and on those attached to rough endoplasmic reticulum (RER) and the nuclear envelope. No calmoduline was observed in the cisternae of RER or the Golgi apparactus. Calmodulin did not appear to be concentrated in the soluble fraction of the cell under the conditions used. Rather, peroxidase reaction product could be seen associated with membranes of the Golgi apparatus the smooth endoplasmic reticulum (SER), and the plasma membrane of both cell bodies and neuronal processes. In the neuronal dendrites, calmodulin appeared to be concentrated on membranes of the SER, small vesicles, and mitochondria. Also, granular calmodulin was observed in the amorphous material. In the synaptic junction, a large amount of calmodulin was seen attached to the inner surface of the postsynaptic membrane, whereas very little was observed in the presynaptic membrane or vesicles. These observations suggest that calmodulin is synthesized on ribosomes and discharged into the cytosol, and that it then becomes associated with a variety of intracellular membranes. Calmodulin also seems to be transported via neuronal processes to the postsynaptic membrane. Calmodulin localization at the postsynaptic membrane suggests that this protein may mediate calcium effects at the synaptic junction and, thus, may play a role in the regulation of neurotransmission.