Activities of the nuclear envelope in the salivary glands of Drosophila

Morphological data are presented concerning the single-membrane-bound vesicles ("oval bodies") associated with the nuclear envelopes of larval salivary gland cells of Drosophila. Data are also presented concerning the existence of cytoplasmic annulate lamellae in these same cells. The mode of formation of these structures, as well as the relationships between them and with other cytoplasmic organelles are described. The possible functional significance of these phenomena is discussed.     

Studies on three mutagen-sensitive mutants of mouse L5178Y cells. II. Complementation analyses between two methyl methanesulfonate-sensitive mutants and between two 4-nitroquinoline-1-oxide-sensitive mutants

Three mutagen-sensitive mutants, MS-1, M10 and Q31, were isolated from mouse L5178Y cells. MS-1 cells are sensitive to methyl methanesulfonate (MMS), M10 cells are cross-sensitive to X-rays, MMS and 4-nitroquinoline-1-oxide (4NQO); and Q31 cells are cross-sensitive to UV and 4NQO. MMS-, X-ray- and UV-sensitive markers in these mutants behaved recessively in hybrids between pairs of these mutants as in hybrids between L5178Y and these mutants as reported before (Shiomi et al., 1982b). Complementation analyses were carried out by forming hybrids between two MMS-sensitive mutants (MS-1 and M10) and between two 4NQO-sensitive mutants (M10 and Q31). MMS and 4NQO survivals were measured in these hybrid cells. MS-1 and M10 were found to belong to different complementation groups for MMS-sensitive phenotypes. The hybrid clones between M10 and Q31 were as sensitive to 4NQO as each of the mutants, indicating codominance of 4NQO sensitivity in these mutants. The hybrids constructed with L5178Y and three mutants were stable as to their chromosome constitution for 100 days of cultivation without selective pressure. From the segregation studies on these hybrids, it is concluded that neither the X-ray-sensitive mutation in M10 nor the UV-sensitive mutation in Q31 is located on the X chromosome.     

Different functions of tight junctions in the ascidian branchial basket

The stigmatal cells in the branchial basket of ascidians from a number of genera have been examined as to the nature and distribution of their intercellular junctions. The branchial wall consists of ciliated and parietal cells; the ciliated cells are arranged in seven rows and are associated by junctions with other cells in the same row as well as with those in adjacent rows. They are also associated by junctions with peripheral parietal cells. Junctions between adjacent ciliated cells in all cases exhibit tight junctions or zonulae occludentes. However, these cell borders also possess fasciae or zonulae adhaerentes if they are in the same row and the ciliary rootlets insert-into these junctions. If the cells are in adjacent rows they exhibit adhaerentes junctions only in species belonging to the orders Phlebobranchiata and Aplousobranchiata. In contrast, if the cells in adjacent rows belong to the order Stolidobranchiata. they never exhibit any adhaerentes junctions and the ciliary rootlets of the basal bodies from the cilia insert instead into the tight junctions and the non-junctional membrane below them. At the homologous junctional borders between adjacent parietal cells and also at heterologous junctional borders between parietal and ciliated cells, tight junctions alone occur, with no co-existing adhaerentes junctions along their lateral borders. Again, fibrils from ciliary rootlets insert into zonulae occludentes. This shows that tight junctions are capable both of forming permeability barriers, in that they can be seen to prevent the entry of exogenous tracers such as lanthanum, and of acting as adhesive devices.     

The mast cells of the mammalian central nervous system. III. Ultrastructural characteristics in the adult rat brain.

The brains of young adult male and female Sprague-Dawley rats were studied with the electron microscope to determine the full ultrastructural picture of two types of perivascular granular cell. One of these, referred to here as the type I cell and described by both light and electron microscopy by several authors, including ourselves, has been reported to be a mast cell (MC) almost identical to MCs outside the CNS. The other, referred to here as the type II cell and described by many authors under almost as many names, was dealt with fully by Ibrahim in several reports and regarded by him as a type of MC. It is felt that the results warrant the conclusions that the type I cells are indeed MCs, while the type II cells are closely allied to the type I cells and probably better adapted to the function they subserve in the CNS of mammals. The similarities between the two cell types probably outnumber the dissimilarities and even these have their counterparts in MCs outside the CNS. The problem of the possible confusion between the type II cells and macrophages, whether reportedly within vessel walls or in the form of modified or special 'pericytic' microglia, is discussed. It is concluded that there is no justification for regarding these cells as macrophages. Because of the similarity between the type II cells and MCs, and because of the high lipid content of the type II cells, it is suggested that these elements be called neurolipomastocytes or neurolipomastocytoid cells.     

The ultrastructure of mouse neuroblastoma cells in tissue culture.

Neuroblastoma cells grown in suspension culture are round and have no distinctive structural characteristics. However, cells transferred to substrates flatten, develop long neurites, and assume the morphology of normal neurons. The resemblance of monolayered neuroblastoma cells to normal neurons is amplified by treatment with hypertonic medium; under these conditions, cell division is inhibited and the neurites become long and differentiated. The treated cells contain clusters of clear vesicles, 400-600 A in diameter, which are morphologically indistinguishable from the synaptic vesicles of normal neurons. Specialized cell contacts are observed between the treated cells as well as between confluent cells grown in normal medium.     

Fine structure of the midgut gland of Phalangium opilio (Chelicerata,Phalangida)

Summary The fine structure of the midgut gland and the changes in composition associated with the digestive activity were examined in Phalangium opilio. In the epithelium four different types of cells are present: ferment cells, resorption cells, and digestion cells which probably turn into excretion cells, as can be seen by many intermediate stages. Ferment cells are found only in the midgut gland and in no other epithelia; therefore they should be regarded as a cell type. The relationship between digestion and resorption cells is not yet clear. No regeneration zone or single regeneration cells could be identified.The ultrastructural changes in these different cells during digestion are described, and their functional aspects are discussed. A hypothetical digestive cycle is constructed from these data. The results are compared with those on other chelicerate midgut glands.     

Similarities and distinctions between murine natural killer cells and lymphokine-activated killer cells

Pretreatment of mice with rabbit anti-asialo GM1 removes both natural killer (NK) effector cells and NK cells responsive to interleukin 2 (IL-2). Spleen cells from these mice do possess normal lymphokine-activated killer (LAK) activity. Young mice (less than 3 weeks of age) do not have NK activity and do not possess IL-2-inducible NK effector cells. Similarly to anti-asialo GM1-treated mice, LAK cells can be generated from these mice. While these experiments indicate clear distinctions between a certain level of NK and LAK precursors, the distinctions are not as clear when analyzing mice congenitally deficient in NK cells. Beige mice which lack NK effector cells and IL-2-inducible NK cells also lack the ability to generate LAK cells. The relationships and differences between NK- and LAK-cell precursors and effectors are discussed.     

Two different forms of gap junctions within the same organism, one with cytoskeletal attachments, in tunicates

The cells of the intestinal tract and the stigmatal cells of the branchial basket have been studied in a range of tunicates including phlebobranch, aplousobranch and stolidobranch ascidians, as well as the doliolid and pyrosomatid thaliaceans. The intercellular gap junctions between gut cells appear conventional in thin section as do those found in the lower part of adjacent stigmatal cells. However, save for the stolidobranchs, the stigmatal cells also have a second kind of gap junction which exhibit an unusual fibrous density in association with their junctional cytoplasmic surfaces; these are found in the apical region of the cells. The fibrous density is particularly well demonstrated in specimens treated with tannic acid during fixation, and subsequent en bloc uranyl acetate staining. In the branchial basket the position of these apical gap junctions is at regular intervals between adhaering junctions, which have a more substantial paramembranous fibrous mat; these two kinds of junctions alternate along deeply undulating membrane appositions. With freeze-fracture, after chemical or cryo-fixation, the gap junctions of the gut and those of the lower part of the stigmatal cells appear typical, with P-face connexons, while in the apical part of cells of the branchial basket the two faces of the gap junctions are very difficult to cleave apart. Frequently the P- and E-faces are found to adhere together in replicas, so that in these apical gap junctional regions, plaques of E-face with pits overlie the PF particles. In addition, regions of cytoplasm, into which the dense fibres project, often cleave over these adhaering E-faces of the apical gap junctions. The presence of these unusual gap junctional features in the apical region of the stigmata in the vicinity of cilia is discussed as regards their functional role.     

Some new findings about Hofbauer cells in the chorionic villi of the human placenta

The cytological structure of the Hofbauer cells was investigated in human placentas of the first and second trimesters of gestation. These cells are found in the stromal channel system of the chorionic villi core. Their walls, which are supported by collagen fiber bundles, are produced by reticulum cells and fibroblasts. The cytoplasmic processes of the Hofbauer cells are in contact with the walls of the channels without being associated with them by desmosomal complexes. Some of these cells have features in common with macrophages, such as cytoplasmic processes, larger vacuoles, many pinocytotic vesicles and intracytoplasmic granules. This system of vacuoles and vesicles enables micropinocytotic activity and phagocytosis. This type of Hofbauer cell resembles the typical macrophages. These cells may play a role in the regulation of stromal water content, transportation of ions and the flow of interstitial fluid. The most original finding of this study are long tubes observed in some Hofbauer cells and extending between the nucleus and the extracellular ground substance through the cytoplasm. One of these tubular formations resembles a cilium in structure with three limiting membranes and is filled with a slightly electron-dense substance. This type of Hofbauer cell may transport information between the nucleus and the extracellular ground substance by means of these tubular structures.     

STUDIES ON THE ENDOPLASMIC RETICULUM : V. Its Form and Differentiation in Pigment Epithelial Cells of the Frog Retina

Pigment epithelial cells of the frog's retina have been examined by methods of electron microscopy with special attention focused on the fine structure of the endoplasmic reticulum and the myeloid bodies. These cells, as reported previously, send apical prolongations into the spaces between the rod outer segments, and within these extensions, pigment migrates in response to light stimulation. The cytoplasm of these cells is filled with a compact lattice of membrane-limited tubules, the surfaces of which are smooth or particle-free. In this respect, the endoplasmic reticulum here resembles that encountered in cells which produce lipid-rich secretions. The myeloid bodies comprise paired membranes arranged in stacks shaped like biconvex lenses. At their margins the membranes are continuous with elements of the ER and in consequence of this the myeloid body is referred to as a differentiation of the reticulum. The paired membranes resemble in their thickness and spacings those which make up the outer segments; they are therefore regarded as intracellular photoreceptors of possible significance in the activation of pigment migration and other physiologic functions of these cells. The fuscin granules are enclosed in membranes which are also continuous with those of the ER. The granules seem to move independently of the prolongations in which they are contained. The report also describes the fine structure of the terminal bar apparatus, the fibrous layer intervening between the epithelium and the choroid blood vessels, and comments on the functions of the organelles depicted.     

Comparative functional analysis of helper T lymphocyte responses to soluble and particulate antigens

An adaptable and sensitive assay to analyze the roles of helper T lymphocytes (TH) which recognize soluble or cell-surface bound antigens in the induction of cytotoxic T lymphocyte precursors (CTLp) is described. Long-term T cell lines that recognize purified protein derivative, keyhole limpet hemocyanin, or Corynebacterium parvum were used in these studies. The ability of T cells from these lines to induce cytotoxic T lymphocyte or antibody responses were compared with their ability to proliferate or release interleukin 2 (IL 2). The results demonstrate that these T cell lines are able to react to soluble antigen by proliferation and IL 2 release. Moreover, the same cell lines are able to interact with CTLp or with the precursors of antibody-secreting B cells to induce a response. In the induction of CTLp we observed an inverse correlation between the number of TH cells required and the concentration of antigen used to pulse the antigen presenting cells. However the correlation between the ability of TH lines to proliferate specifically in response to antigen and to act as helpers for CTLp and B cells was not absolute as cells with compromised proliferative capacity were able to efficiently deliver inductive signals.     

Contacts between endocrine and exocrine cells in the pancreas

Summary Close contacts between exocrine and endocrine cells were observed in human and rat pancreas. The presence of junctional specializations, including desmosomes, tight and gap junctions, as well as interdigitations between endocrine and exocrine cells, implies that these cells are structurally and functionally associated.     

Cytoneme-mediated cell-to-cell signaling during development

Cell-to-cell communication is vital for animal tissues and organs to develop and function as organized units. Throughout development, intercellular communication is crucial for the generation of structural diversity, mainly by the regulation of differentiation and growth. During these processes, several signaling molecules function as messengers between cells and are transported from producing to receptor cells. Thus, a tight spatial and temporal regulation of signaling transport is likely to be critical during morphogenesis. Despite much experimental and theoretical work, the question as to how these signals move between cells remains. Cell-to-cell contact is probably the most precise spatial and temporal mechanism for the transference of signaling molecules from the producing to the receiving cells. However, most of these molecules can also function at a distance between cells that are not juxtaposed. Recent research has shown the way in which cells may achieve direct physical contact and communication through actin-based filopodia. In addition, increasing evidence is revealing the role of such filopodia in regulating spatial patterning during development; in this context, the filopodia are referred to as cytonemes. In this review, we highlight recent work concerning the roles of these filopodia in cell signaling during development. The processes that initiate and regulate the formation, orientation and dynamics of cytonemes are poorly understood but are potentially extremely important areas for our knowledge of intercellular communication.     

Regulation of mouse oocyte growth: probable nutritional role for intercellular communication between follicle cells and oocytes in oocyte growth

Intercellular communication, as determined by two different assay procedures, was established in vitro between mouse oocytes free of adhering follicle cells and monolayers of either follicle or 3T3 cells. Both of these cell types are known to be able to form homologous gap junctions, and follicle cells naturally form heterologous gap junctions with oocytes in vivo. Monolayers of L cells that are communication deficient did not establish intercellular communication with oocytes as determined by the two different assays for intercellular communication. The diameter of oocytes cultured for 4 days in medium or on monolayers of L cells decreased markedly, 9.7 and 13.1 micron, respectively. In contrast, oocytes cultured for 4 days on follicle cell monolayers increased on the average about 4.7 micron in diameter. Oocytes cultured for 4 days on monolayers of 3T3 cells decreased slightly in diameter, i.e., 2.1 micron. Results from these experiments support a nutritional role for intercellular communication between follicle cells and oocytes in oocyte growth.     

Atypical gap junctions in the ciliary epithelium of the albino rabbit eye

Gap junctions are present between nonpigmented and pigmented epithelial cells layers, as well as between the adjacent pigmented cells, but not between the nonpigmented epithelial cells which face the posterior chamber. The unusual feature of these gap junctions is a dark banding which regularly appears every three rows of subunits. This dark band is the equivalent width of two rows of facets.     

Caveats: numerical requirements in graph theory based quantitation of tissue architecture.

Graph theory based methods represent one approach to an objective and reproducible structural analysis of tissue architecture. By these methods, neighborhood relations between a number of objects (e.g., cells) are explored and inherent to these methods are therefore certain requirements as to the number of objects to be included in the analysis. However, the question of how many objects are required to achieve reproducible values in repeated computations of proposed structural features, has previously not been adressed specifically. After digitising HE stained slides and storing them as grey level images, cell nuclei were segmented and their geometrical centre of gravity were computed, serving as the basis for construction of the Voronoi diagram (VD) and its subgraphs. Variations in repeated computations of structural features derived from these graphs were related to the number of cell nuclei included in the analysis. We demonstrate a large variation in the values of the structural features from one computation to another in one and the same section when only a limited number of cells (100-500) are included in the analysis. This variation decreased with increasing number of cells analyzed. The exact number of cells required to achieve reproducible values differ significantly between tissues, but not between separate cases of similar lesions. There are no significant differences between normal and malignantly changed tissues in oral mucosa with respect to how many cells must be included. For graph theory based analysis of tissue architecture, care must be taken to include an adequate number of objects; for some of the structural features we have tested, more than 3000 cells.     

STRUCTURAL DEVELOPMENT OF THE CAUDICLE OF AN ORCHID (EPIDENDRUM)

Structural changes during development of the caudicle of Epidendrum ibaguense (Orchidaceae) were followed using light and electron microscopy. The caudicle begins as a mass of small meristematic cells in the microsporangium. The central cells of the mass enlarge and enter a division cycle. Completion of this cycle produces linear tetrads of cells which remain attached to each other throughout the life of the caudicle. These tetrads then accumulate a thick secondary wall on their outer surfaces. These thick-walled cells form the main structural element of the caudicle at maturity. The cells on the periphery of the mass differentiate into a gradation of cell types characterized by their very high content of smooth ER. The majority of these thin-walled cells are located at the junction points between adjacent caudicles and between caudicle and pollinium. The cells in these positions produce large quantities of a lipid polymer. This product is synthesized in association with large tubular ER complexes present in discrete areas in the cytoplasm. Plastids of these cells also produce a lipidic substance. Some thin-walled cells are located among the thick-walled structural cells; however, the lipid polymer is not synthesized in these cells. At maturity, all thin-walled cells undergo breakdown and autolysis resulting in gradual obliteration of cytoplasmic detail and organelles. In the lipid containing thin-walled cells, lysis of the cells releases the lipidic masses which then form the elastic joining compound between adjacent caudicles and between caudicles and pollinia.     

Identification of K(+) channels in the plasma membrane of maize subsidiary cells

The stomatal complex of Zea mays consists of two guard cells with the pore in between them and two flanking subsidiary cells. Both guard cells and subsidiary cells are important elements for stoma physiology because a well-coordinated transmembrane shuttle transport of potassium and chloride ions occurs between these cells during stomatal movement. To shed light upon the corresponding transport systems from subsidiary cells, subsidiary cell protoplasts were enzymatically isolated and in turn, analyzed with the patch-clamp technique. Thereby, two K(+)-selective channel types were identified in the plasma membrane of subsidiary cells. With regard to their voltage-dependent gating behavior, they may act as hyperpolarization-dependent K(+) uptake and depolarization-activated K(+) release channels during stomatal movement. Interestingly, the K(+) channels from subsidiary cells and guard cells similarly responded to membrane voltage as well as to changes in the K(+) gradient. Further, the inward- and outward-rectifying K(+) current amplitude decreased upon a rise in the intracellular free Ca(2+) level from 2 nM to the micro M-range. The results indicate that the plasma membrane of subsidiary cells and guard cells has to be inversely polarized in order to achieve the anti-parallel direction of K(+) fluxes between these cell types during stomatal movement.