Junctional complex revisited by high-resolution scanning electron microscopy

The present study correlates the ultrastructural morphology of junctional complexes as revealed by transmission electron microscopy (TEM) with that observed by high-resolution scanning electron microscopy (HRSEM), thanks to a new modification of the osmium tetroxide maceration technique. The removal of all cytoplasmic organelles by this technique allows the inspection of the inner side of the plasmalemma. With this treatment, a continuous band of tightly packed particles is observed at the most apical portion of lateral membranes. Just below this band, irregular clusters of apparently identical particles are placed all around the cellular contour. The topographical correspondence among these clusters and spot desmosomes seen by TEM identifies them as desmosomes. The continuous band seems to represent the combination of both zonulae, occludens and adherens. Regarding the nature of the particles, we suppose that they probably consist of peripheral membrane proteins clustered at the cytoplasmic surface of intercellular junctions and involved in the linkage between cytoskeleton and plasmalemma.     

Ultrastructure of Paneth cells in the intestine of various mammals

Paneth cells in the following species were observed under an electron microscope: human, rhesus monkey, hare, guinea pig, rat, nude rat, mouse, golden hamster, and insect feeder bat. Secretory granules containing homogeneous electron-dense materials were observed in the Paneth cells of humans, monkeys, hares, guinea pigs, and bats; mouse Paneth-cell granules were bipartite (central core and peripheral halo), and the Paneth cells in rats and golden hamsters had secretory granules showing various electron densities. In humans, monkeys, and bats, immature granules near the Golgi apparatus sometimes showed bipartite substructure. The number and size of secretory granules were also diverse among various animal species. Some lysosome-like bodies were commonly observed in peri- or supranuclear regions, though the size and shape of the bodies differed from cell to cell. In apical cytoplasm, small clear vesicles (100–200 nm diameter) were more-or-less observed in all species examined, and it was especially note that rat Paneth cells contained many clear vesicles. Small dense-cored vesicles (150–200 nm diameter) were rare. It is unlikely that the various ultrastructural features of Paneth cells correlate with the phylogenetical classification.     

Role of the golgi apparatus in cellular pathology

The Golgi apparatus response to pathological disorders is predominantly as an intermediary component of membrane biogenesis where it is involved in processing, sorting and secretion of materials via secretory granules, and in the formation of lysosomes. A common initial response of the Golgi apparatus to any stress is an alteration or cessation of secretory activity. In the transformed cell, the Golgi apparatus is altered both morphologically and biochemically, suggesting a shift from a secretory to a membrane-generating mode of functioning. However, since fewer or less well-developed Golgi apparatus are frequently found in transformed cells, analytical methods of membrane isolation developed for normal tissues may not always yield equivalent results when applied to tumors. Cell surface alterations characteristic of malignant cells may result from modifications occurring at the level of the Golgi apparatus. Some lysosomal dysfunctions may result from underglycosylation of acid hydrolases by the Golgi apparatus. The use of cell-free systems between endoplasmic reticulum and Golgi apparatus or within Golgi apparatus cisterane is providing a new approach to the elucidation of the role of the Golgi apparatus in normal as well as pathological states.     

Tales of tethers and tentacles: golgins in plants

As plant Golgi bodies move through the cell along the actin cytoskeleton, they face the need to maintain their polarized stack structure whilst receiving, processing and distributing protein cargo destined for secretion. Structural proteins, or Golgi matrix proteins, help to hold cisternae together and tethering factors direct cargo carriers to the correct target membranes. This review focuses on golgins, a protein family containing long coiled-coil regions, summarizes their known functions in animal cells and highlights recent findings about plant golgins and their putative roles in the plant secretory pathway.     

Retaining ionic concentrations during in vitro storage of tissue for microanalytical studies

When human or animal tissue is to be investigated by X-ray microanalysis, it is sometimes necessary to store the tissue between removal from the organism and freezing. However, when excised tissue is stored in buffer, the elemental concentrations in the cell may change. In the present study, it was attempted to develop a storage buffer that would retain the cellular elemental concentrations close to their in situ values. To start, the NaCl component in Krebs–Ringer buffer was exchanged for K-gluconate and KCl for NaCl. This buffer was called a ‘100% high K⁺ solution’. Starting from this solution, part of the K-gluconate was replaced by an equivalent amount of NaCl. Incubation of excised rat liver (4 °C, 4 h) in 85% high K⁺ solution resulted in retention of cellular Na, K, Ca, S and Mg concentration most closely to the in situ state, whereas cellular Cl was retained best when the tissue was incubated in 75% high K⁺ solution. For rat submandibular gland, incubation in 80% high K⁺ solution resulted in optimal retention of cellular Na, K, Ca, P, S and Mg, while Cl was retained best in a 70% high K⁺ solution. Based on these results, an optimally modified Krebs–Ringer solution for the liver would consist of 119 mM K⁺, 26 mM Na⁺ and 45 mM Cl^?. An optimally modified Krebs–Ringer bicarbonate solution for the submandibular gland would be composed of 96 mM K⁺, 53 mM Na⁺ and 46 mM Cl^?. After incubation in the modified solutions (at 4 °C), cellular Na, Mg, S, Cl, K and Ca in both tissues were maintained close to the in situ state throughout a 6-h incubation. The cellular P concentration was reduced after incubation for 1 h; thereafter, in the liver cells it remained at this lower level for the rest of the incubation, whereas in the submandibular gland tissue it increased again after 4 h. The increase in cell volume (oedema) was less in tissue stored in the modified solutions, than in the 100% high K⁺ solutions. Incubation in high Na⁺ buffers (4 °C, 6 h) resulted in a progressive increase in the percentage of cells showing trypan blue uptake. A similar increase in trypan blue uptake was seen in the modified solution, but this increase levelled off after 4 h. After cholinergic stimulation in high Na⁺ solution (25 °C, 1 min), the expected decrease in cellular Cl concentration was seen in submandibular gland cells that had previously been preserved (4 °C, 4 h) in the modified solution, but not in those that had been preserved in the 100% high K⁺ solution.     

Differential morphometric values induced in Golgi apparatus of higher plant cells by aldehyde and permanganate fixation

In order to determine the best conditions to carry out quantitative ultrastructural studies in plant specimens, five different fixation techniques, including some of the most reported electron microscopy fixatives (glutaraldehyde-paraformaldehyde, osmium tetroxide, potassium permanganate), were assayed in onion root meristems to check their ability to induce morphometric changes in Golgi apparatus ultrastructure. Although the parameters evaluated showed in all cases the same tendency, values obtained after permanganate fixation were always higher than those found after aldehyde techniques (especially aldehyde-osmium). Aldehyde followed by osmium fixation appears as the most indicated fixation method when accurate quantitative ultrastructural studies are to be developed.     

Perisynaptic Schwann cells of the vertebrate motor endplate bear modified cilia

Perisynaptic Schwann cells (PSCs), descendants of the myelinating Schwann cells, cover the axon terminal of the vertebrate motor endplate of the skeletal muscle fiber. PSCs are assumed to support the function of the axon terminal. This function suggests a net material transport in the direction of the axon terminal. Morphologically it is to be expected that these cells have a cytoskeleton aligned to the axon terminal. Investigations clarifying this statement have not yet been undertaken. From previous investigations we know, however, that the PSCs have a microtubule-organizing center, which is a part of this cytoskeleton. The centrioles of the organizing center may also participate in the formation of a modified cilium structure whose function is unknown. In the present investigation, characteristic ultrastructural features of the modified cilium structure and its relationship to the Golgi apparatus and the axon terminal are presented. A function for the modified cilium structure is discussed.     

Electron microscopic study of the morphology and morphogenesis of virus of hemorrhagic fever with renal syndrome

The morphology and morphogenesis of the virus of hemorrhagic fever with renal syndrome (HFRS) and the associated ultrastructural changes in neurons of the infected mouse brain were examined by electron microscopy. The primary location of the infection in large neurons was in the Golgi apparatus, which had highly proliferated laminar and vesicular profiles. A small number of matured virus particles were found later individually or in small groups within the distended Golgi cisternae and vesicles. Most of the virus particles were round, oval, or elongated and measured about 70–110 nm in diameter. A lipid bilayered viral envelope with an external fringe of surface projections could be resolved at high magnification. The maturation (budding) of the virus occurred exclusively at smooth membrane vesicles, and predominantly at membranes in, or adjacent to, Golgi cisternae. Viral inclusion bodies containing fine filamentous material were seen frequently in close proximity to sites of virus maturation. The known morphological and morphogenetic characteristics of the virus particles observed in infected mouse brain gave further evidence for taxonomic identification of HFRS virus as a member of the family of Bunyaviridae.     

Trans-Golgi reticulum

The trans-Golgi apparatus reticulum is that portion of the Golgi apparatus located in the trans-most aspect of the stack exhibiting certain characteristic morphological and functional characteristics. The membranes of the trans-Golgi reticulum are reticular in form, thickened with plasma membrane-like characteristics and with a considerable portion of their surface covered by clathrin coats. The enzymes thiamine pyrophosphatase and sialyl- and galactosyl transferases are functional markers. Correlative studies show the trans-Golgi apparatus reticulum to be involved in glycoprotein, enzyme and receptor processing and sorting along multiple pathways. Sorting and transfer of constituents to lysosomes, to secretory granules, or to the plasma membrane emerge as dominant functions.     

ERES (ER exit sites) and the "secretory unit concept"

The higher plant Golgi apparatus consists of hundreds of individual Golgi stacks which move along the cortical ER, propelled by the actomysin system. Anterograde and retrograde transport between the endoplasmic reticulum (ER) and the plant Golgi occurs over a narrow interface (around 500 nm) and is generally considered to be mediated by COP-coated vesicles. Previously, ER exit sites (ERES) have been identified on the basis of to localization of transiently expressed COPII-coat proteins. As a consequence it has been held that ERES in higher plants are intimately associated with Golgi stacks, and that both move together as an integrated structure: the "secretory unit". Using a new COPII marker, as well as YFP-SEC24 (a bona fide COPII coat protein), we have made observations on tobacco leaf epidermis at high resolution in the CLSM. Our data clearly shows that COPII fluorescence is associated with the Golgi stacks rather than the surface of the ER and probably represents the temporary accumulation of COPII vesicles in the Golgi matrix prior to fusion with the cis-Golgi cisternae. We have calculated the numbers of COPII vesicles which would be required to provide a typical Golgi-associated COPII-fluorescent signal as being much less than 20. We have discussed the consequences of this and question the continued usage of the term "secretory unit".