Establishment of epidermal cell columns in mammalian skin: computer simulation

The present investigation shows an example of tissue formation in relation to individual cell properties. Epidermis of mammalian skin has recently been shown to be organized during ontogeny into neat vertical cell columns, in which a cell approximates the flattened form of Kelvin's tetrakaidecahedron, a 14-sided body with eight hexagonal faces and six square faces. Such an epidermal architecture is compatible with the organization and turnover of stacked cells, a constant loss of surface cells and a supply that cells from a basal layer differentiate during migration to the surface. The developmental process from irregular cell aggregate into neat columns in skin is simulated on a digital electronic computer with the assumption as follows: a new cell migrating upward from a basal layer jostles and settles at the less crowded area among upper cells. After migration of many cells to the surface, the simulation demonstrates that cells have been stacked in neat columns, and the stability of the architecture is also shown.     

A computer simulation of cell stacking for even thickness in mammalian epidermis

The method of even stacking of epidermal cells in mammalian skin was studied by computer simulation. The epidermis consists of neat vertical columns of stacked, flattened, tetrakaidecahedral cells. Cells which have been proliferated in a basal layer migrate upwards, occupy the bottom regions of vertical columns, and become members constituting columns. Computer simulations demonstrated that the column height becomes considerably varied if the cells are randomly supplied from the basal layer. In contrast, if the cells are assumed to have an ability to find the uppermost region among the column's bases consisting of one base where the cell has reached and its neighbouring bases, the cells stack into columns whose heights are remarkably uniform even if the cells are randomly supplied. The results indicated that an epidermal structure consisting of the flattened polyhedral cells could itself function as a control mechanism of the epidermal thickness.     

Organized Protoplasmic Units of the Plant Cell: I. THEIR OCCURRENCE, ORIGIN, AND STRUCTURE

The development of the placental tissue of the tomato fruithas been investigated and it has been found that the protoplasmof cells of this tissue separates into organized units of protoplasmas the fruit matures and ripens. Similar organized protoplasmicunits have been detected in other fruits at a comparable stageof development. A study has been made of the structure and stability of theseorganized units, and it has been deduced that these organizedunits of protoplasm are in fact compartments of the protoplasmof the cell which have become separated from one another. Eachorganized protoplasmic unit is able to exist independently ofthe cell for several days, and is apparently surrounded by amembrane of the endoplasmic reticulum. The possible significance of these new findings is discussedin relation to the structural and biochemical organization ofthe plant cell.     

Die epidermale Columnärstruktur

Zusammenfassung Die strukturelle Organisation von Hornschicht und nichtverhornter Epidermis des Mäuseohres wurde an FITC-gefärbten, säurenachbehandelten Schnitten sowie an Aufsichtspräparaten untersucht und mit Meerschweinchen- und menschlicher Epidermis verglichen. Bei der für eine dünne Epidermis charakteristischen Zellsäulenbildung entstehen 5–6eckige Hornzellen, die sich nur geringgradig und regelmäßig überlappen. In der Hornschicht wie in der columnär geordneten lebenden Epidermis ist die Überlappung (oder Stufung) die Folge eines abwechselnden Einrückens differenzierender Zellen in die Säulen. Dabei lassen sich 2 Formen der Zellbewegung abgrenzen: a) das seitliche Einschieben unterhalb der Nahtstelle zweier Zellsäulen und b) das senkrechte Aufwandern subcolumnärer Basalzellen. Ein unterschiedliches Zellalter scheint für die Bildung von Zellsäulen erforderlich zu sein.

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The columnar structure of the epidermisPossible mechanism of differentiation

Summary The structural organisation of the Stratum corneum and the nonkeratinized epidermis of the mouse ear has been analyzed with a fluorescence-acid-staining technique and was compared to human and guinea pig epidermis. In cell columns, which are typical for a thin epidermis, 5–6-sided horny cells overlap each other regularly and to a constant extent. In the horny layer as well as in the stacked portion of the living epidermis this overlap results from an alternatant movement of differentiating cells into the stacks. Two forms of cellular movement can be observed: sideward slipping into the column below the junction of two cell stacks and vertical upward movement of subcolumnar basal cells. The age-dependent differences of non-stacked cells seem to be one of the factors necessary for the organization of thin mammalian epidermis.

Für die finanzielle Unterstützung dieser Arbeit sei der Deutschen Forschungsgemeinschaft gedankt.     

Overview of the structure of all-AT oligonucleotides: organization in helices and packing interactions

We present the crystalline organization of 33 all-AT deoxyoligonucleotide duplexes, studied by x-ray diffraction. Most of them have very similar structures, with Watson-Crick basepairs and a standard average twist close to 36 degrees. The molecules are organized as parallel columns of stacked duplexes in a helical arrangement. Such organization of duplexes is very regular and repetitive: all sequences show the same pattern. It is mainly determined by the stacking of the terminal basepairs, so that the twist in the virtual TA base step between neighbor duplexes is always negative, approximately -22 degrees. The distance between the axes of parallel columns is practically identical in all cases, approximately 26 A. Interestingly, it coincides with that found in DNA viruses and fibers in their hexagonal phase. It appears to be a characteristic distance for ordered parallel DNA molecules. This feature is due to the absence of short range intermolecular forces, which are usually due to the presence of CG basepairs at the end of the oligonucleotide sequence. The duplexes apparently interact only through their diffuse ionic atmospheres. The results obtained can thus be considered as intermediate between liquid crystals, fibers, and standard crystal structures. They provide new information on medium range DNA-DNA interactions.     

Ultrastructure and development of the rhabdomeric eyes in Lineus viridis (Heteronemertea, Nemertea)

Nemerteans are undoubtedly members of the Spiralia, although their phylogenetic relationships are still a matter of debate. The apparently acoelomate organization suggests a relationship with the platyhelminths, whereas the blood-vascular system has been interpreted as an equivalent to coelomic cavities of annelids, indicating a close relation between annelids and nemerteans. Like other spiralians, most nemertean species are known to have one or several pairs of rhabdomeric and subepidermally situated eyes when adult. The development of these eyes as well as the mode in which the eyes are multiplied is as yet unknown. This is the first attempt to investigate eye formation in a nemertean. In the heteronemertean Lineus viridis (Müller, 1774) the everse rhabdomeric eyes are located deeply underneath the epidermis and consist of a few pigment cells that form a cup-like structure with interdigitating processes that contain numerous pigment granules. In hatchlings, the optical cavity contains processes of 12 sensory cells, each bearing a single cilium and various microvilli. The perikarya of these cells are located distally from the pigment cup. During further development the number of cells increases. Eye development starts with a small anlage situated underneath the epidermis, irrespective of whether this is the first eye or any additional one. The anlage consists of five unpigmented cells and three dendritic processes, each bearing apical microvilli and a single cilium. There is no evidence for an epidermal origin of the eyes. In L. viridis eye formation resembles that described in platyhelminths in which eyes also develop as cerebral derivatives. Although this result has the potential to influence the discussion on the position of Nemertea, the data have to be interpreted with care, since development of L. viridis is derived within the Nemertea.     

Eisosome proteins assemble into a membrane scaffold

Spatial organization of membranes into domains of distinct protein and lipid composition is a fundamental feature of biological systems. The plasma membrane is organized in such domains to efficiently orchestrate the many reactions occurring there simultaneously. Despite the almost universal presence of membrane domains, mechanisms of their formation are often unclear. Yeast cells feature prominent plasma membrane domain organization, which is at least partially mediated by eisosomes. Eisosomes are large protein complexes that are primarily composed of many subunits of two Bin-Amphiphysin-Rvs domain-containing proteins, Pil1 and Lsp1. In this paper, we show that these proteins self-assemble into higher-order structures and bind preferentially to phosphoinositide-containing membranes. Using a combination of electron microscopy approaches, we generate structural models of Pil1 and Lsp1 assemblies, which resemble eisosomes in cells. Our data suggest that the mechanism of membrane organization by eisosomes is mediated by self-assembly of its core components into a membrane-bound protein scaffold with lipid-binding specificity.     

Expression of keratin K14 in the epidermis and hair follicle: insights into complex programs of differentiation

Keratins K14 and K5 have long been considered to be biochemical markers of the stratified squamous epithelia, including epidermis (Moll, R., W. Franke, D. Schiller, B. Geiger, and R. Krepler. 1982. Cell. 31:11-24; Nelson, W., and T.-T. Sun. 1983. J. Cell Biol. 97:244-251). When cells of most stratified squamous epithelia differentiate, they downregulate expression of mRNAs encoding these two keratins and induce expression of new sets of keratins specific for individual programs of epithelial differentiation. Frequently, as in the case of epidermis, the expression of differentiation-specific keratins also leads to a reorganization of the keratin filament network, including denser bundling of the keratin fibers. We report here the use of monospecific antisera and cRNA probes to examine the differential expression of keratin K14 in the complex tissue of human skin. Using in situ hybridizations and immunoelectron microscopy, we find that the patterns of K14 expression and filament organization in the hair follicle are strikingly different from epidermis. Some of the mitotically active outer root sheath (ORS) cells, which give rise to ORS under normal circumstances and to epidermis during wound healing, produce only low levels of K14. These cells have fewer keratin filaments than basal epidermal cells, and the filaments are organized into looser, more delicate bundles than is typical for epidermis. As these cells differentiate, they elevate their expression of K14 and produce denser bundles of keratin filaments more typical of epidermis. In contrast to basal cells of epidermis and ORS, matrix cells, which are relatively undifferentiated and which can give rise to inner root sheath, cuticle and hair shaft, show no evidence of K14, K14 mRNA expression, or keratin filament formation. As matrix cells differentiate, they produce hair-specific keratins and dense bundles of keratin filaments but they do not induce K14 expression. Collectively, the patterns of K14 and K14 mRNA expression and filament organization in mitotically active epithelial cells of the skin correlate with their relative degree of pluripotency, and this suggests a possible basis for the deviation of hair follicle programs of differentiation from those of other stratified squamous epithelia.     

Synchronized oscillation in a modular neural network composed of columns

The columnar organization is a ubiquitous feature in the cerebral cortex. In this study, a neural network model simulating the cortical columns has been constructed. When fed with random pulse input with constant rate, a column generates synchronized oscillations, with a frequency varying from 3 to 43 Hz depending on parameter values. The behavior of the model under periodic stimulation was studied and the input-output relationship was non-linear. When identical columns were sparsely interconnected, the column oscillator could be locked in synchrony. In a network composed of heterogeneous columns, the columns were organized by intrinsic properties and formed partially synchronized assemblies.     

Synchronized oscillation in a modular neural network composed of columns

The columnar organization is a ubiquitous feature in the cerebral cortex. In this study, a neural network model simulating the cortical columns has been constructed. When fed with random pulse input with constant rate, a column generates synchronized oscillations, with a frequency varying from 3 to 43 Hz depending on parameter values. The behavior of the model under periodic stimulation was studied and the input-output relationship was non-linear. When identical columns were sparsely interconnected, the column oscillator could be locked in synchrony. In a network composed of heterogeneous columns, the columns were organized by intrinsic properties and formed partially synchronized assemblies.     

Dynamic nature and function of epidermal Langerhans cells in vivo and in vitro: a review, with emphasis on human Langerhans cells.

Summary Epidermal Langerhans cells (LC) are Birbeck granule-containing bone-marrow-derived cells, which are located mainly in the suprabasal layer of the epidermis. They can be readily identified by their strong expression of CDIa and MHC class II molecules. In addition to these classical properties, an extensive phenotypic profile of normal human LC, summarized in this review, is now available. The powerful capacity of LC to activate T lymphocytes is clearly documented and, to date, LC are recognized as the prominent antigen-presenting cells of the skin immune system. They are generally believed to pick up antigens encountered in the epidermis and to migrate subsequently from the epidermis to the skin-draining lymph nodes. Upon arrival in the paracortex of lymph nodes, the antigen-laden LC transform into interdigitating cells and they present antigen to naive T lymphocytes in a MHC class II-restricted fashion; this results in the generation of antigen-specific immune responses. It has also been demonstrated that transformation of LC into interdigitating cells occurs when LC are culturedin vitro. Bothin vivo andin vitro studies have indicated that properties of LC, such as phenotype, morphology and the stimulatory potential to activate T lymphocytes, are dependent on the local microenvironment in which the LC reside. The essential role of LC in the induction of contact allergic skin reactions and skin transplant rejection is well established.     

The Preferred Directions of Conjunctive Grid X Head Direction Cells in the Medial Entorhinal Cortex Are Periodically Organized

The discovery of speed-modulated grid, head direction, and conjunctive grid x head direction cells in the medial entorhinal cortex has led to the hypothesis that path integration, the updating of one’s spatial representation based on movement, may be carried out within this region. This hypothesis has been formalized by many computational models, including a class known as attractor network models. While many of these models propose specific mechanisms by which path integration might occur, predictions of these specific mechanisms have not been tested. Here I derive and test a key prediction of one attractor network path integration mechanism. Specifically, I first demonstrate that this mechanism predicts a periodic distribution of conjunctive cell preferred directions in order to minimize drift. Next, I test whether conjunctive cell preferred directions are in fact periodically organized. Results indicate that conjunctive cells are preferentially tuned to increments of 36°, consistent with drift minimization in this path integration mechanism. By contrast, no periodicity was observed in the preferred directions of either pure grid or pure head direction cells. These results provide the first neural evidence of a nonuniform structure in the directional preferences of any head direction representation found in the brain.     

Liquid crystalline ordering of nucleosome core particles under macromolecular crowding conditions: evidence for a discotic columnar hexagonal phase.

Macromolecular crowding conditions occurring inside the cell nucleus were reproduced experimentally with solutions of mononucleosome core particles to study their supramolecular organization. We report here that under these conditions, and over a large range of monovalent salt concentrations, mononucleosome core particles self-assemble to form a discotic liquid crystalline phase characterized in polarizing and freeze-fracture electron microscopy. Mononucleosomes are stacked on each other to form columns, which are themselves closely packed into an hexagonal array. The nucleosome concentration, estimated from the network parameters, falls in the range of values measured in cell nuclei. We suggest that these concentrated solutions, although their organization cannot be immediately compared to the organization of chromatin in vivo, may be used to investigate the nucleosome-nucleosome interactions. Furthermore, this approach may be complexified to take into account the complexity of the eucaryotic chromatin.     

Laminar processing in the visual cortical column

Sensory regions of neocortex are organized as arrays of vertical columns composed of cells that share similar response properties, with the orientation columns of the cat's visual cortex being the best known example. Interest in how sensitivity to different stimulus features first emerges in the columns and how this selectivity is refined by subsequent processing has fueled decades of research. A natural starting point in approaching these issues is anatomy. Each column traverses the six cortical layers and each layer has a unique pattern of inputs, intrinsic connections and outputs. Thus, it makes sense to explore the possibility of corresponding laminar differences in sensory function, that is, to examine relationships between morphology and physiology. In addition, to help identify general patterns of cortical organization, it is useful to compare results obtained from different sensory systems and diverse species. The picture that emerges from such comparisons is that each cortical layer serves a distinct role in sensory function. Furthermore, different cortices appear to share some common strategies for processing information but also have specialized mechanisms adapted for the demands of specific sensory tasks.     

An epidermal proliferative unit-like structure in the epithelium of mouse bladder observed by backscattered electron imaging

Summary The simultaneous use of a silver-staining technique, backscattered electron imaging and stereo-tilts has made it possible to visualize the spatial distribution of cell nuclei in the stretched epithelium of the bladder of mice. This study has led to the observation that a structural organization resembling the epidermal proliferative unit, previously found in the skin exists also in bladder epithelium. However, the proliferative unit in the bladder was different in that it contained a higher number of cells per unit, and an absence of columns of inactive squamous cells. These findings may indicate that epidermal proliferative unit-like structures represent a common form of organization in some epithelia.     

Self-assembly of thin plates from micrococcal nuclease-digested chromatin of metaphase chromosomes

The three-dimensional organization of the enormously long DNA molecules packaged within metaphase chromosomes has been one of the most elusive problems in structural biology. Chromosomal DNA is associated with histones and different structural models consider that the resulting long chromatin fibers are folded forming loops or more irregular three-dimensional networks. Here, we report that fragments of chromatin fibers obtained from human metaphase chromosomes digested with micrococcal nuclease associate spontaneously forming multilaminar platelike structures. These self-assembled structures are identical to the thin plates found previously in partially denatured chromosomes. Under metaphase ionic conditions, the fragments that are initially folded forming the typical 30-nm chromatin fibers are untwisted and incorporated into growing plates. Large plates can be self-assembled from very short chromatin fragments, indicating that metaphase chromatin has a high tendency to generate plates even when there are many discontinuities in the DNA chain. Self-assembly at 37°C favors the formation of thick plates having many layers. All these results demonstrate conclusively that metaphase chromatin has the intrinsic capacity to self-organize as a multilayered planar structure. A chromosome structure consistent of many stacked layers of planar chromatin avoids random entanglement of DNA, and gives compactness and a high physical consistency to chromatids.     

Keratinization of fish skin with special reference to the catfish Bagarius bagarius

Summary Histochemical reactions indicating keratinization have previously been demonstrated in parts of the epidermis of Bagarius bagarius. Fluorescence histochemistry and electron microscopy have now confirmed these results. Elevated areas of the epidermis are capped by a layer of dead cells with altered contents. On the outer aspect of these cells a dense layer, 18 nm thick, beneath the plasma membrane corresponds to the resistant envelope found in keratinized cells in tetrapod vertebrates. In Bagarius this layer does not extend to all faces of the keratinized cells, but a similar envelope has been detected in two other sites of piscine keratinized epidermis investigated, namely in the breeding tubercles of Phoxinus phoxinus and in the teeth of Lampetra fluviatilis. In the elevated areas of Bagarius-epidermis, the epithelial cells undergo progressive changes in cytoplasmic organization as they become more superficial. The second tier from the surface is sealed by tight junctions and is separated from the overlying keratinized cells by a sub-corneal space resembling that found in keratinized amphibian epidermis. Histochemical evidence of a high lipid content in the outer layers of the epidermis correlates with the presence of lipid inclusions and lamellated membranous profiles in the material studied by electron microscopy. Histochemical results show that the fin skin of Blennius pholis is not keratinized, but secretes a cuticle, histochemically reactive for both proteins and glycoproteins.     

Localization of light-harvesting complex II to the occluded surfaces of photosynthetic membranes

The photosynthetic membranes of green plants are organized into stacked regions interconnected by nonstacked regions that have been shown to be biochemically and structurally distinct. Because the stacking process occludes the surfaces of appressed membranes, it has been impossible to conduct structural or biochemical studies of the outer surfaces of the photosynthetic membrane in regions of membrane stacking. Although stacking is mediated at this surface, it has not been possible to determine whether membrane components implicated in the stacking process, including a major light-harvesting complex (LHC-II), are in fact exposed at the membrane surface. We have been able to expose this surface for study in the electron microscope and directly label it with antibodies to determine protein exposure. The appearance of the newly exposed outer stacked surface highlights the extreme lateral heterogeneity of the photosynthetic membrane. The surface is smooth in contrast to the neighboring nonstacked surface that is covered with distinct particles. Although some investigators have suggested the existence of a cytochrome b6/f-rich boundary region between stacked and nonstacked membranes, our results provide no structural support for this concept. To explore the biochemical nature of the occluded membrane surface, we have used an mAb against the amino terminal region of the LHC-II. This mAb clearly labels the newly exposed outer stacked surface but does not label the inner surface or the outer nonstacked surface. These experimental results confirm the presence of the amino terminal region of this complex at the outer surface of the membrane in stacked regions, and also show that this complex is largely absent from nonstacked membranes.     

The epidermal and inner epithelial lining of the operculum inClarias batrachus (Clariidae,Siluriformes)

The operculum may be divided into the proximal, the distal and the intermediate regions. The epithelium lining the inner surface of the operculum (EISO) and the opercular epidermis (OE) of these regions inClarias batrachus show significant differences in their thickness, in the density, in the dimensions of mucous cells and club cells, and in the distribution of lymphocytes, melanocytes, taste buds and ampullary organs. These differences in structural organization are associated with the different conditions prevailing at these locations. Rich vascularization observed in the tissues underlying the OE has been correlated with assisting the fish in accessory respiration. In contrast, the tissues underlying the EISO are poorly vascularized. Accessory respiration in this region may not be so advantageous.     

Self organization of the microtubule network. A diffusion based model

Microtubules form organized polymer networks in cells. Experimental evidence indicates that their mechanical properties do not play a significant role in the generation of such regular patterns. This spatial organization seems closely related to their dynamic behavior. Here we use mathematical modeling to define conditions under which microtubular dynamics result in self organization. We demonstrate that random diffusional processes can generate regular microtubule organizations under specified kinetic conditions which are found to be compatible with the known properties of tubulin polymers. The organizing forces are the tubulin concentration gradients which are generated by the polymer growth. The present analysis has been restricted to the phase of establishment of the microtubule network. The same conceptual framework, applied to steady state pattern maintenance suggests that the kinetic requirements for self organization might ultimately be responsible for such extraordinary in vivo microtubule dynamics, as the rapid turnover and “dynamic instability” of the interphase network.