The association of microtubules with the plasmalemma in epidermal tendon cells of the river crab

The mode of association of microtubules (MTs) with the plasmalemma in epidermal tendon cells of the river crab, Potamon dehaani was studied by thin-section electron microscopy. In the leg muscle, the tendon cells connect striated muscle cells with the cuticle, forming specialized junctions at both ends. At the muscle-tendon cell junction, the apposed plasmalemmas are interdigitated in a zig-zag pattern separated by a uniform space of about 50 nm, where the basal lamina is shared by two cells. At the tendon cell-cuticle junction, the plasmalemma of the tendon cell forms many conical invaginations, into which dense fibrous material extends from the cuticle. Inside the tendon cell, numerous microtubules run parallel to the direction of tension transmission and are arranged into parallel bundles of various sizes. Within such bundles, fine filamentous structures cross-link adjacent MTs. MTs span the entire length of the cell and attach at their both ends to the junctional domains of the plasmalemma. The junctional plasmalemma is characterized by formation of an electron-dense undercoat, through which MTs are connected with the plasmalemma proper. The ultrastructural features of MT association with the plasmalemma are basically the same at both junctions. At the junctions, MTs usually terminate with free ends and are linked laterally to the plasmalemmal undercoat with fine filamentous structures. These observations emphasize the role of the plasmalemmal undercoat as a device of the attachment of MTs to the plasmalemma.     

A mechanism for rapid transport of colloidal particles by flounder renal epithelium

Large quantities of colloidal particles were rapidly transported around the junctional complex into the lateral intercellular spaces by flounder renal epithelial cells. Large invaginations containing particles developed in the apical cytoplasm of cells when tracer particles were injected into the tubular lumens. Some membranebounded profiles containing particles appeared close to the lateral intercellular spaces. Particles were then found in the lateral intercellular spaces, between the basal plasmalemma and the basement membrane, and within the basement membrane. It is suggested that this transport might operate in situ and provide a morphological mechanism to explain a type of protein transport noted in the renal tubules of another flounder species by Maack and Kinter ('67). It is interesting to consider that perhaps a similar mechanism for the transport of intact proteins might also operate in mammalian nephrons as well.     

Ultrastructure of the integumental melanophores of the Australian lungfish,Neoceratodus forsteri

The integumental melanophores of Australina lungfish, Neoceratodus forsteri, were examined by light and electron microscopy and found to possess essentially the same structural characteristics observed in other vertebrates. The epidermal melanophores are located in the intermediate epidermis and possess round perikarya and slender dendrites extending into nearby intercellular spaces. The dermal melanophores are found immediately below the basement membrane as well as in the deeper dermis. These cells possess flattened nuclei and dendrites running parallel to the basement membrane. Each melanophore contains numerous oval or elliptical, intensely electron-dense melanosomes, relatively large mitochondria, systems of vacuolar endoplasmic reticulum, groups of free RNP particles, and some microfilaments. Only a few, short microtubules could be demonstrated in the perinuclear cytoplasm of the dermal melanophore, while a relatively large number of late premelanosomes are found both in perikarya and dendritic processes of epidermal melanophores. These premelanosomes exhibit a particulate internal structure in cross section. Both melanosomes and premelanosomes occur singly in the cytoplasm of epidermal cells, thereby confirming the existence of the epidermal melanin unit in the lowest vertebrates thus far examined electron microscopically.     

The formation of intercellular spaces in the cotyledons of developing and germinating pea seeds

Summary Electron microscopic observations have shown that the intercellular spaces in the storage parenchyma of the cotyledons of pea (Pisum sativum L.) seeds arise schizogenously. The wall segments adjoining future intercellular spaces display a triple zonation and contain regions with electron-dense material. Space formation starts at the central point of contact between several cells and spreads then further up to the electron-dense, intra-wall structures. As a result of this the electron-dense material is found again in the corners of intercellular spaces. It is proposed that the intra-wall structures may have an important function in limiting the schizogenous process. The localization of the intercellular spaces is thus predetermined. The amount of electron-dense material in their corners increases considerably during further development of the embryo. During germination wall segments between two intercellular spaces diverge resulting in a fusion of several spaces.     

Histochemical and ultrastructural analysis of developing adipocytes in the fetal pig

Histochemical and ultrastructural aspects of adipocyte differentiation in subcutaneous tissue of fetal pigs were analyzed in a longitudinal study. A matrix of collagen fibers surrounding adipocytes developed after the establishment of a distinct and continuous PAS-positive basement membrane. The degree of plasma membrane invagination and specialization was positively correlated with the extent of basement membrane and collagen matrix formation. Close spatial relationships between narrow, smooth endoplasmic reticulum, plasma membrane invaginations, the surface of lipid droplets and mitochondria were observed in differentiating adipocytes. Histochemical and ultrastructural criteria for the identification of preadipocytes are: (1) perivascular location; (2) mitochondria localized in the Golgi zone; (3) cytosolic glycogen; (4) rough endoplasmic reticulum with cisternae uniformly and approximately 600 A wide; (5) free ribosomes and few polysomes, and (6) lipid droplets encased by microfilaments. These criteria permitted clear distinction from obvious fibroblasts and macrophages. Other stromal cells were morphologically abnormal. Occasionally, adipocytes and perivascular cells exhibited close intercellular contacts that were morphologically distinct from intercellular contacts between contiguous endothelial cells.     

The morphology of the gills of the freshwater African crab Potamon niloticus (Crustacea: Brachyura: Potamonidae): A scanning and transmission electron microscopic study

The gills of the African freshwater crab Potamon niloticus -Ortmann have been investigated by scanning and transmission electron microscopy. Potamon has seven pairs of phyllobranchiate gills contained in the branchial chambers. From the central axis of the gills arise bilaterally situated thin flaps, the lamellae. The afferent branchial vessel (the epibranchial vessel) is located on the dorsal aspect of the gill arch and the efferent vessel (the hypobrancial vessel) on the ventral side. Between these two blood vessels, the blood percolates through the lamellar vascular channels where it is oxygenated. The lamellae consist of an epithelial cell layer covered by a thin cuticle which consists of tightly fused but distinct layers. The epithelial cells approach each other at regular intervals and fuse in the middle of the lamellar sinus delineating the vascular channels. Apical profuse membranous infoldings and numerous mitochondria characterize the epithelial cells, features typical of cells involved in active transport of macro- and micromolecules. In Potamon , however, there were no distinct gas exchange and osmoregulatory regions of the gills. On average, the cuticle was 0.78 μm thick while the epithelial cell was 6 μm. Cells that were morphologically similar to the renal glomerular podocytes of the vertebrates were observed in the efferent gill vessel of Potamon. These cells have been said to be phagocytic and may play an important defensive role in the crustaceans. Although basically the morphology of the gills of Potamon is similar to that of the other decapods, fine structural differences were evident as would be intuitively expected in a group of animals that has undergone such remarkable adaptive radiation.     

The surface characteristics of the plasma membrane of the exocrine pancreas.

Regional differentiation of the plasma membrane and related structures of the exocrine pancreas has been studied ultrastructurally and cytochemically. Fixation with an osmium tetroxide-silver acetate solution produced abundant fine precipitates on the luminal and basal surface of the centroacinar but not the acinar cells. Staining with dialyzed iron (DI) revealed the heaviest concentration of anionic sites on the luminal plasma membrane of the acinar cells, including the surface of both the intercellular canaliculi and the main lumen. The reactive sites on the apical acinar plasmalemma appeared to consist of discrete globules. DI-reactivity of the lateral basal membranes was most prominent in the centroacinar cells and essentially absent in the acinar cells but was weak relative to that of the acinar-cell apical plasmalemma. The lamina lucida of the basement membrane of the duct stained with DI, but that of basement membrane under acinar cells did not. Sialidase digestion prior to DI staining abolished the staining of plasma membranes. These results indicate that duct epithelial cells, including most prominently the centroacinar cells, are chiefly responsible for electrolyte and fluid transport.     

Structural organization of ovarian follicle cells in the cotton bug (Dysdercus intermedius) and the honeybee (Apis mellifera)

Summary The somatic epithelia of Dysdercus and Apis follicles were analyzed by electron microscopy, and the patterns of F-actin and microtubules were studied by fluorescence microscopy. The epithelia in both species differ considerably in shape and in the organization of the cytoskeleton. During previtellogenic stages, the epithelium consists of columnar-shaped cells with small (Dysdercus) or no (Apis) lateral intercellular spaces. During vitellogenesis, the follicle cells round up; the intercellular spaces increase in size in Dysdercus follicles, whereas in Apis follicles they remain small. Along the basal surface of the follicle cells, there are conspicuous parallel bundles of microfilaments perpendicular to the anteroposterior axis of the follicles. In the honeybee, these microfilament bundles are present in long filopodia, most of which are embedded in thickenings of the basement membrane and extend over the surfaces of neighbouring cells. In the cotton bug, the basal surface of the follicle cells is thrown into parallel folds. The microfilament bundles are located just underneath the cell membrane where the folds contact the basement membrane. In the polar regions of the Dysdercus follicle, the epithelial cells become flat and adhere to each other without forming intercellular spaces. The basement membrane is particularly thick in the polar areas; this has also been observed in Apis follicles around the intercellular bridge connecting oocyte and nurse cells.     

Arrangement of connective tissue components in the walls of seminiferous tubules of man and monkey.

In primates the membrane separating the seminiferous epithelium from the interstitial space is composed of one to three (monkey) or two to six layers (man) of myoid cells associated with one to two layers of fibrocyte-like adventitial cells. All these cells are separated from each other by irregular spaces filled with various connective tissue intercellular components. Subjacent to the elements of the seminiferous epithelium is a continuous, often redundant, basement membrane. A similar basement membrane-like material forms a layer next to and over small areas of the plasma membrane of myoid cells. Collagen fibrils grouped in bundles of various sizes are seen in all connective tissue layers but are particularly abundant in the space between the seminiferous epithelium and the innermost layer of myoid cells. Elastic fibrils demonstrated by the Verhoeff iron hematoxylin technique are also present. Composed of a homogeneous material, the elastic fibrils are short, irregular, branching entities with a diameter comparable to or smaller than that of collagen fibrils. In addition, an abundance of microfibrils with a diameter of 12-15 nm is present in the various connective tissue layers. These microfibrils have a densely stained cortex and a lightly stained core. When seen close to the myoid cells, bundles of micro fibrils appear to insert on well defined areas next to the plasma membrane. These areas commonly face the patches of electron-dense material observed on the inner aspect of the plasma membrane of the myoid cells and in which the actin filaments are inserted. Bundles of microfibrils often span the gap between myoid cells of the same layer as well as those of adjacent layers. Microfibrils are also closely related to the surface of elastic fibrils and are seen intertwining with collagen fibrils. Thus microfibrils appear to bridge and bind together adjacent myoid cells and anchor the surface of these cells to the bundles of elastic and collagen fibrils present in the intercellular spaces of the limiting membrane.     

Transmission of symbiotic algae through sexual reproduction in hydra: movement of algae into the oocyte

The histological pathway by which intracellular symbiotic Chlorella move into the developing oocytes of hydra was investigated at the ultrastructural level. Algae move from within the digestive cells of the endoderm to within the oocytes of the ectoderm in a three-step process. First, the algae are released by digestive cells into the mesolamella (basement membrane). Second, the algae move as individual cells into the adjacent intercellular spaces of the ectoderm. Third, they are taken up by the oocyte by phagocytosis. This transfer occurs only in the central regions of the ovary, and only after oocytes have reached an advanced stage. Normally the mesolamella is separated from the ectodermal interstitial spaces by a layer of epitheliomuscular cell muscle processes. This layer degenerates in the region where algae will move into the ectoderm. This study shows that algae move as individual cells and not intracellularly within processes of the epithelial cells.     

Potassium pyroantimonate and osmium-zinc iodide reactivity in the tubular epithelium of the opisthonephric kidney of the sea lamprey,Petromyzon marinus L

Pyroantimonate precipitate indicates that the epithelium of the proximal tubule is the only segment of the tubular nephron of the fresh water lamprey where large accumlations of cations are distributed. Unusually large amounts of reaction product are located within the lateral intercellular spaces and within vesicles closely associated with the plasma membrane at the lateral and basal surfaces. This technique suggests the continuity of these vesicles with the plasma membrane and alludes to the possibility of an endomembranous system of vesicles and the intercellular spaces as vehicles for ion transport. Lateral intercellular spaces of proximal tubules of lower vertebrates may play a different role in kidney function that their counterparts in higher vertebrates. Osmium-zinc iodide has a specificity for certain cells within the proximal, intermediate, and distal segments, but no structural differences are noted when these cells are compared to unstained cells. Smooth endoplasmic reticulum remains unstained in the distal segment but the stain has a strong affinity for elements of the Golgi apparatus, lysosomes, and the nuclear envelope of all cell types. This technique does not suggest a structural or functional similarity between cells of the distal segment and the chloride cells of the gills of teleosts.     

Does sulphide detoxication occur in the gills of the hydrothermal vent shrimp, Rimicaris exoculata?Y a-t-il détoxication des sulfures dans les branchies de la crevette hydrothermale, Rimicaris exoculata ?

Ultrastructural observations of the gills of the hydrothermal vent shrimp Rimicaris exoculata reveal that the epithelial cells contain numerous mitochondria clustered around unusual organelles (diameter of 0.7 to 2.5 microns) containing membrane stacks. These organelles were termed sulphide-oxidising bodies (SOBs) by structural analogy with organelles observed in the tissues of species adapted to sulphide-rich environments. Moreover, in the gills of R. exoculata, mitochondria display numerous electron-dense granules in their stroma. Such ultrastructural features suggest that sulphide detoxication may probably occur in the gills of R. exoculata. Comparable structures were also described in the gills of other hydrothermal vent species, as the alvinellid Pompeii worms that, as R. exoculata, are housing ectosymbiotic bacteria.     

Coelomic Cells in the Connective Tissue Spaces of the Clitellar Epithelium of Lumbricus friendi Cognetti, 1904 (Oligochaeta): an Ultrastructural Study

The ultrastructure of the connective tissue spaces in the clitellar epithelium has been studied in the earthworm Lumbricus friendi. Four morphological types of coelomic cells are described: amoebocytes, mucocyte-like cells, pigment cells and crystal-containing cells. The amoebocytes are characterized by the presence of spherical to oval electron-dense granules, phagocytic vacuoles and numerous microtubules located in the Golgi areas. The mucocyte-like cells show the features of the mucocytes reported in enchytraeid worms (globular inclusions with filamentous and homogeneous, moderately electron-dense material, as well as a filopodous process). The pigment cells contain typical spindle-shaped osmiophilic granules, microtubules (not reported before) and glycogen particles. The crystal-containing cells show inclusions which are polygonal in section with a striated substructure (periodicity of about 4.5 nm). Apart from the mucocyte-like cells, the coelomocytes showed cytoplasmic processes attached to the basement membrane of the spaces. The possible functions of these cells are discussed and a common peritoneal origin is postulated on the base of their morphological and cytological features.     

Some ultrastructural observations on the mode of association of microtubules with the plasmalemma in epidermal tendon cells of the river crab

Summary— The ultrastructural aspects of the association of microtubules (MTs) with the plasmalemma in epidermal tendon cells of the river crab, Polamon dehaani, were studied by thin-section electron microscopy combined with detergent treatment. In the tendon cell, MTs were linked laterally by anchoring filaments to the plasmalemma via a submembranous electron-dense layer called the plasmalemmal undercoat. To further clarify how such anchoring filaments are spatially related to the plasmalemma through the undercoat, we carefully examined and compared thin-section images obtained from various specimen preparations using saponin and Triton X-100. When the tissues were treated with saponin or Triton, electron-dense materials in the undercoat were extracted in varying degrees to expose internal substructures. The undercoat appeared to show a two-layer organization, the inner and outer layers. In more extracted samples, filamentous networks became prominent in the outer layer. Anchoring filaments were seen to attach to such filamentous networks, which in turn were linked to the plasmalemma proper. Thus, it may be reasonable to consider that the filamentous network constitutes the core structure of the plasmalemmal undercoat which is structurally reinforced by extractable electron-dense materials.     

Early stages of nodulation in roots of Oxytropis arctobia (Leguminosae) induced by the arctic rhizobial strain N31

Early stages of nodulation of roots of the arctic legume Oxytropis arctobia by the arctic rhizobial strain N31 was characterized under controlled conditions. Root hair deformation occurred as a result of inoculation of seedlings. Infection threads were seen invading target cells in the root cortex of the newly formed nodule tissue. In emergent nodules, bacteria remained surrounded by a moderately electron-dense matrix, within the intercellular space. The target cells were rich in lipid bodies, proplastids, mitochondria and polyribosomes. Fibrillar material, microfilaments and small vesicles were present at the point of bacterial release, where the infection thread was devoid of its wall. Bacteria were found to be encircled by plasmalemma invaginations forming "symbiosomes". Lipid bodies were present near the membrane of the infection thread, close to the site of bacterial release, and in close association with plasmalemma.     

Ultrastructure of spermatogenesis in the sea star,Asterina minor

The ultrastructural features of spermatogenesis were investigated in the hermaphroditic sea star Asterina minor. The primordial germ cells in the genital rachis contain small clusters of electron-dense material (nuage material) and a stack of annulate lamellae. They also have a flagellum and basal body complex situated close to the Golgi complex. After the development of the genital rachis into the ovotestis, spermatogenic cells increase in number and differentiation begins. Nuage material is observed in spermatogonia, but it gradually disappears in spermatocytes. The annulate lamellae do not exist beyond the early spermatogonial stage. By contrast, a flagellum and basal body complex are found throughout spermatogenesis. The Golgi-derived proacrosomal vesicles appear in the spermatocyte and coalesce to form an acrosomal vesicle in the early spermatid. The process of acrosome formation is as follows: (1) a lamella of endoplasmic reticulum (ER) continuous with the outer nuclear membrane encloses the posterior portion of the acrosomal vesicle; (2) the vesicle attaches to the cell membrane with its anterior portion; (3) periacrosomal material accumulates in the space between the acrosomal vesicle and the ER; (4) the nucleus proper changes its features to surround the acrosome; (5) amorphous, electron-dense material is deposited under the electron-dense disk; and (6) the nucleus forms a hollow opposite the electron-dense material.     

An electron microscopic study of absorption of peroxidase-conjugated immunoglobulin G by guinea pig visceral yolk sac in vitro.

In the guinea pig and some other animals, passive immunity is conferred on the developing fetus by passage of immunoglobulin from mother to fetus across the yolk sac. In order to examine the cytological pathway involved in immunoglobulin transport, guinea pig visceral yolk sacs from late in gestation were exposed in vitro to peroxidase-conjugated guinea pig immunoglobulin G (IgG-HRP). Tissue was then fixed, incubated to show the site of localization of peroxidase reaction product and prepared for electron microscopy. The results suggested that the first step in the uptake of IgG-HRP by yolk sac is attachment of the protein to the surface coats of endocytic invaginations at the apical surfaces of the endodermal cells. The endocytic vesicles then appear to pinch off from the surface and move deeper into the cytoplasm. Some of the small endocytic vesicles fuse with large apical vacuoles, which often contain large amounts of reaction product. Other small endocytic vesicles pinch off from the surface, move deeper into the cytoplasm and fuse with the lateral plasmalemma; their protein content is emptied into the intercellular space by exocytosis. From the intercellular spaces the protein presumably diffuses across the basement membrane and connective tissue spaces and enters the vitelline capillary bed. It is postulated that the latter cellular pathway, involving small vesicles and the intercellular spaces, is utilized by those immunoglobulins which are transferred intact across the yolk sac endoderm.     

Extracellular matrix of the human thymus: immunofluorescence studies on frozen sections and cultured epithelial cells

The immunohistochemical detection of elements of the human thymic extracellular matrix in situ and in vitro is described. In the normal thymus, the intracapsular and intraseptal fibers were strongly labeled by anti-type I collagen antiserum. Basement membranes bordering the capsule, septae, and perivascular spaces were intensely stained by anti-type IV collagen, anti-fibronectin, and anti-laminin sera. In hyperplastic myasthenia gravis thymuses, the major changes consisted of discontinuities of the basement membrane adjacent to clusters of epithelial (keratin-containing) cells, among which an unusual connective framework (densely labeled by all the antisera) was observed. In vitro, most epithelial cells were strongly labeled by antifibronectin serum and to a lesser extent by the anti-type IV collagen and anti-laminin sera. In addition, fibronectin, laminin, and type IV collagen were detected in the intercellular spaces bordering the epithelial cells in culture. Results show that thymic epithelial cells participate in the synthesis of extracellular matrix elements, which as a result of their localization and influence on epithelial cell growth, should be regarded as constitutive components of the thymic microenvironment.     

F-Actin and Tubulin During Spermatogenesis in Gamasid Mites (Acari: Parasitina)

Abstract F-actin and tubulin behaviour was investigated using fluorescence probes and electron microscopy in the course of spermatogenesis in two gamasid mites, Porrhostaspis lunulata Müller (Parasitidae) and Pergamasus truatellus Athias-Henriot (Pergamasidae). In spermatogonia and primary spermatocytes of both species, the proteins were localized mainly in the intercellular bridges and, in lesser quantities, in the cytoplasm. Overall, actin was present along the plasma-lemmal contact sites of the gonial cells. At the beginning of spermatid elongation, actin could be detected in two regions: in perinuclear cytoplasm and under the plasmalemma. Subplasmalemmal actin, visible as threads running along acrosome-adhering protrusions of the nuclear envelope, is supposedly located within the electron-dense material filling the subacrosomal gap. Tubulin was found on both sides of each actin thread; its location was consistent with two sets of microtubules adhering to the inner acrosomal membrane. Their involvement in acrosome shaping is suggested. As spermatid elongation terminated, the previous pattern of proteins disappeared. In Pergamasus, however, actin emerged briefly near the centrifugal ends of spermatids (granular bodies zone). In spermatocyte-containing cysts, actin and tubulin fluorescence (more pronounced in Porrhostaspis) was associated with intercellular junctions between the cyst cells. In both species, diffuse actin fluorescence was also detected in the cytoplasm of cyst cells assembling elongated spermatids; the reaction was intensified at the end of the elongation process, when the cytoplasm of cyst cells aggregated around the centripetal ends of spermatids.     

Staining of proteoglycans in mouse lung alveoli. I. Ultrastructural localization of anionic sites

Summary In order to contrast anionic sites, in mouse lung alveoli, two staining procedures were applied: (a) staining with Ruthenium Red and Alcian Blue and (b) staining with Cuprolinic Blue in a critical electrolyte concentration method. The Ruthenium Red-Alcian Blue staining procedure revealed electron-dense granules in the alveolar basement membrane. The granules were closely associated with the epithelial cell membrane and continued to stain even when the procedure was carried out at a low pH, indicating the presence of sulphate groups in the granules.After staining with Cuprolinic Blue, electron-dense filaments, also closely associated with the cell membrane, became visible in the basement membrane of type I epithelial cells. Their length depended on the MgCl₂ concentration used during staining. At 0.4m MgCl₂, the length was mostly within the range 100–180 nm. Using a modified Cuprolinic Blue method, the appearance of the filaments closely resembled that of spread proteoglycan monomers with their side-chains condensed. The basement membrane of type II epithelial cells also contained filaments positive towards Cuprolinic Blue; their length, however, was smaller in comparison with those of type I epithelial cells. The filaments lay in one plane and provided the whole alveolus with an almost continuous sheet of anionic sites. Cuprolinic Blue staining also revealed filaments in the basement membrane of the capillary endothelial cells. Furthermore, Cuprolinic Blue-positive filaments (average length about 40 nm) became apparent in close contact with collagen fibrils and separated from each other according to the main banding period of the collagen fibrils (about 60 nm), indicating a specific ultrastructural interaction between these two components. Filaments connecting collagen fibrils with each other were also detected.