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Biomedical subjects

A Reichenbach

Publications and source records attributed to A Reichenbach.

At least 127 records · Page 7Linked to original sources
PubMed1991

Development of the rabbit retina. IV. Tissue tensility and elasticity in dependence on topographic specializations.

A method is introduced for the quantification of specific compliance and the elasticity of small pieces of living retinal tissue. These pieces are fixed at their margins by means of tissue glue, and loaded with a small iron spherule the bending force of which can be gradually enhanced by the action of an electromagnet. Retinal bending caused by such calibrated forces is measured by a horizontal light microscope, and used for estimations of specific compliance and elasticity of the tissue. Three different particular regions of the rabbit retina--periphery, visual streak, and (prospective) medullary rays--were tested at several post-natal developmental stages. From very early stages on (day 2 p.p.) up to adulthood the peripheral retina was found to be significantly more tensile than the two other central regions. This can be shown to depend greatly on the thickness of the tissue which is lower in the retinal periphery. During early post-natal development, all retinal regions except the (prospective) medullary rays become thinner. The tensility of the tissue increases, with the exception of the medullary rays which reduce their compliance strongly. In the adult retina, however, the tensility of all retinal regions is reduced as compared with the neonatal tissue. This seems to be caused by a constant gradual increase of the elasticity of the retina during development which, in turn, may be caused by several developmental parameters, e.g. the formation of synapses, the outgrowth of glial side branches ensheathing neighbouring neuronal cells, or a reduction in extracellular clefts. It is proposed that these differences in tensility between different retinal regions, may be the cause for differential retinal expansion driven by the intraocular pressure. Thus, simple mechanical features of the tissue may contribute to the formation of important topographic specializations of the retina, e.g. the visual streak as the site of highest visual acuity.

Aging↗
PubMed1990

Rabbit retinal Müller cells in cell culture show gap and tight junctions which they do not express in situ.

Retinae of early postnatal rabbits were enzymatically dissociated and explanted in a culture system. The prospective myelinated region was discarded in order to avoid the presence of astrocytic or mesenchymal cells. After about 14 days in vitro (DIV), outgrowing glial (Müller) cells formed what light optically appeared to be confluent monolayers but by electron microscopy was shown to consist of flat epithelioid cells which overlapped considerably by extension of cytoplasmic tongues. Applying the freeze-fracture technique, apposed membranes of these cells were demonstrated to express infrequently but consistently both gap and tight junctions. This kind of junctions has never been observed on the membrane of rabbit Müller cells in situ. In comparison with Müller cell membranes in situ, the density of intramembrane particles was considerably reduced. Orthogonal arrays of particles which are characteristic elements of Müller cells in situ were not detected. Our results suggest that in homogeneous cell culture, Müller cells form some kind of epithelium-like specialized intercellular junctions. This situation resembles that of closely related glial cell types which form homogeneous layers in situ as e.g. retinal pigment epithelium cells expressing tight junctions, and marginal astrocytes being coupled by extensive gap junctions.

Animals↗
PubMed1990

Orthogonal arrays of intramembranous particles in the Müller cell and astrocyte endfoot membrane of rabbit retina. Postnatal development and adulthood.

The freeze-fracture technique was applied to the retina of early postnatal and adult rabbits to investigate the distribution and density of orthogonal arrays of intramembrane particles (OAP) within the vitread endfoot membranes. In adult animals, two distinct types of endfoot membranes were observed within the central myelinated retina but not in the retinal periphery. One type of endfoot membranes contained low density of individual 'background' particles, and a more or less stripe-like pattern of OAP; this type was found only within the myelinated centre and is concluded to represent membranes of retinal astrocytes. The other type of endfoot membranes was rather tightly packed with individual 'background' particles, and contained OAP which formed rows only at the margins of footplates; this kind of membrane was found throughout the whole retina, and represents Müller cell endfeet. The density of OAP in both types of endfoot membranes was higher within the central myelinated retina than in the Müller cell endfeet of the retinal periphery. In early postnatal retinae, a discrimination between the two types of endfeet was impossible. At the day of birth, only very few OAP were observed, and the majority of footplates were free of OAP. Within the next 10 days, both density and size of OAP were found to increase but fail to reach adult levels. Quantitative data are presented with the hope of providing a basis for future correlation with functional maturation of rabbit retinal glia.

Animals↗
PubMed1990

Membrane ultrastructure preservation and membrane potentials after isolation of rabbit retinal glial (Müller) cells by papain.

Enzymatically isolated retinal glial (Müller) cells have been the subject of many electrophysiological studies. Local high membrane conductivities for potassium ions have been speculated to correspond with local occurrence of orthogonal arrays of intramembranous particles (OAP) observed in freeze-fracture replicas of retinal Müller cells in situ. We studied whether such OAP are preserved after enzymatic digestion of the retinal tissue which is necessary for isolation of living cells for electrophysiology. We found that strong papain digestion leads not only to disturbances in the cell's ultrastructure as seen in ultrathin sections but evokes both a redistribution of intramembranous particles and a disappearance of OAP as seen in the freeze-fracture replica. Furthermore, such isolated cells have low membrane potentials and lose their topographical specialization in K+ conductance. If, however, the retinae were exposed to papain as short as possible to get just some isolated cells, their cytoplasmic and membranous ultrastructure was preserved very well, and high resting membrane potentials were recorded in cells with marked regional specialization of membrane conductivity. Our results show that indeed sites of high K+ conductance may correspond with the occurrence of OAP, even in isolated cells.

Animals↗
PubMed1990

Determination of the extracellular tortuosity in nuclear layers of the central nervous system by resistance measurements on a geometrical model.

The electrical resistivity of nuclear layers within CNS tissues is simulated by a densely packed array of glass spherules with a thin silver layer on their surfaces. Various mean spherule diameters and different thicknesses of the silver layer are tested; the relations between spherule diameter and thickness of silver mantle are in the same range as those between cell soma diameters and width of the extracellular clefts. Measuring the contribution of the silver layers to the total volume--corresponding to the extracellular volume fraction alpha within CNS tissues--and the resistance of columns of silvered spherules, tortuosity factors lambda 2 are calculated. Means of lambda 2 are found to vary within 5.6 and 9.2, independent of both spherule diameters and thickness of the silver layer, but strongly dependent on the packing density of the spherules. This latter dependence is described by a simple formula. These results are used to calculate the resistivities of the nuclear layers of the retina and of the granular layer in the hippocampal area dentata, based on morphometric data gained from own studies and literature reports, respectively. It is shown that such layers with densely packed cells express very high resistivity because of both low extracellular volume fraction and high tortuosity. Implications for current source density analyses, and for pathological events like epileptogenesis, are discussed. In an appendix, an analytical solution of the problem is given for the case of a cubic array of surface-conducting spheres.

Animals↗
PubMed1990

Primary neurulation in teleosts--evidence for epithelial genesis of central nervous tissue as in other vertebrates.

Early teleostian embryos were studied by scanning electron microscopy. Transversal and longitudinal views of the neural anlage clearly demonstrate that it has an ordinary epithelial organization consisting of parallel columnar cells. Unless it has been supposed earlier, there is no solid thickened neural plate but the neural ectoderm is tightly folded forming a very narrow neural groove. Thus, primary neurulation in teleosts is shown to occur similar to that in other vertebrates viz. by folding of the neural plate to get the neural tube. That means that the neuroepithelial cells retain their polarity instead to become organized from a randomly oriented mass of unpolarized cells, as it is thought to occur in secondary neurulation.

Animals↗
PubMed1990

Radial glial cells are present in the velum medullare of adult monkeys.

As a rule, in thin mammalian CNS tissues such as the median eminence and the retina, radial glia remains the dominating macroglia in adulthood, whereas in most other regions of the brain radial glial is substituted by multipolar macroglia i.e. astroglia. The Velum medullare is another thin CNS tissue but there are no reports on the dominating macroglia forms of this structure. Thus, Golgi-impregnated sections of adult monkey brains were studied for the presence of radial glial cells. Indeed, this structure was found to be transversed by many radial glial fibres terminating with pial endfeet whereas in adjacent thick brain tissues the glia limitans was formed by marginal astrocytes. It is concluded that fibrous radial glia may dominate in adult mammalian and even primate CNS tissues with a thickness of up to 1 mm.

Animals↗
PubMed1989

Cell type-specific distribution of cathepsin B and D immunoreactivity within the rabbit retina.

The cellular localization of cathepsin B and D immunoreactivity was demonstrated at the light microscopic level in the retina of adult rabbits by use of the peroxidase-antiperoxidase technique. Antisera were raised against rat liver enzymes. Whereas cathepsin D immunoreactivity was confined to Müller (glial) cells, cathepsin B was demonstrated in some, but not all, neuronal cell types. It is proposed that the two enzymes might carry different functions within the neuronal versus glial compartment.

Animals↗
PubMed1989

Glia:neuron index: review and hypothesis to account for different values in various mammals.

The present paper proposes a hypothesis to account for different values of the glia:neuron index in comparable central nervous system tissues of various mammals. This hypothesis assumes that K+ ions released by active neurons are a mitogenic signal for glial cells. The thicker the tissue (for example, the brain wall), the more difficult is efficient K+ clearance, and more perinatal glial cell proliferation should occur. Thus, this hypothesis accounts for higher glia:neuron indices in mammals with thicker brain walls.

Aging↗
PubMed1989

Size and density of glial and neuronal cells within the cerebral neocortex of various insectivorian species.

Morphometric measurements were done on frontal sections through the somatosensory neocortex of various insectivorian species. All measured parameters varied with the size of animals; there was a better correlation with the ventriculartopial brain wall thickness than with the brain weight. The following rules were evaluated: with increasing brain wall thickness, 1) lamina I becomes thinner; 2) the nuclei of both neuronal and glial cells become larger; 3) the volume density of neuronal cells decreases greatly; 4) the volume density of glial cells increases slightly; and 5) as a result, the glia:neuron index increases markedly. There was no equal number of neurons under a unit surface area in the cortices of any species studied. Developmental processes that might account for the above-mentioned rules are discussed in this report.

Animals↗
PubMed1989

Attempt to classify glial cells by means of their process specialization using the rabbit retinal Müller cell as an example of cytotopographic specialization of glial cells.

The rabbit retinal Müller cell is one of the most widely studied glial cell types, and it has all forms of contacts that a glial cell can express, viz. 1) to a (ventricular) fluid space, 2) to a mesenchymal borderline (basal lamina), and 3) to neuronal compartments. This cell demonstrates the local adaptation of cell processes to the microenvironment with which they are in contact. Summarizing available data on Müller cells and other glial cell types, it is concluded that the structure with which the process is in contact determines the type of glial cell process that develops. The type I process has microvilli, desmosome-like junctions, and high Na+,K+-ATPase activity; this type of process is in direct contact with a fluid such as cerebrospinal fluid. The type II endfoot-bearing process contains gliofilaments and has a high K+ conductivity; this type of process is covered by a basal lamina and is in contact with mesenchyme. The type III sheath-bearing process insulates neuronal compartments and expresses suitable membrane properties for glia-neuronal communication. Since structurally similar processes have been shown to have similar physiological properties, a new systematic classification of glial cells is proposed, based on the presence or absence of defined types of cell processes. This approach is believed to provide new insights into the function of neuroglia in both the central and peripheral nervous systems, in vertebrates and invertebrates, and even during ontogenetic development.

Animals↗
PubMed1989

The structure of rabbit retinal Müller (glial) cells is adapted to the surrounding retinal layers.

Radial glial (Müller) cells of the rabbit retina were studied by various techniques including Golgi impregnation, scanning electron microscopy, horseradish peroxidase application, and staining of enzymatically isolated cells. This combination of methods produced detailed information on the specialized morphology of the Müller cells within the different topographical regions of the retina, and of the Müller cell processes within the various retinal layers. As a general rule, the retinal periphery contains short thick Müller cells with big endfeet, whereas the thick central retina is occupied by long slender cells with small endfeet. Independent of their location within the retina, Müller cell processes were found to be adapted to the structure of the surrounding retinal layers. Within the outer and inner nuclear layers, Müller cell processes (and somata) extend thin cytoplasmic "bubbles" ensheathing the neuronal somata, as do the "velate" astrocytes in the brain. In the plexiform layers, Müller cells extend many fine side branches between the neuropil, comparable to the protoplasmic astrocytes of the brain. In the thick myelinated nerve fibre layer of the central retina the Müller cell processes are rather smooth, similar to those of fibrous astrocytes. It is concluded that the neuronal microenvironment determines the morphology of a given glial process, or even of a part of a glial process running through a specialized neuronal compartment.

Animals↗
PubMed1989

Potassium as a signal for both proliferation and differentiation of rabbit retinal (Müller) glia growing in cell culture.

Retinal glial (Müller) cells were grown from explants of early postnatal rabbit retinae. The resulting monolayers of flat cells were exposed to control media (containing 5.85 mM K+), and to media with enhanced K+ concentrations (10 and 20 mM) or arginine-vasopressin (AVP, 20 micrograms/ml) or epithelial growth factor (EGF, 10 ng/ml). Autoradiographically, protein synthesis was quantified as L-[3H]-lysine incorporation, and DNA synthesis as [3H]-thymidine incorporation. Furthermore, the activity of Na+,K(+)-ATPase was measured radiochemically. Short exposure to either moderately enhanced K+ concentrations (10 mM) or to AVP, stimulated L-[3H]-lysine incorporation into the cells. Long-lasting exposure to either high K+ concentrations (20 mM) or to EGF stimulated [3H]-uptake. The Na+,K(+)-ATPase activity of cell cultures increased with increasing K+ concentration of the media. It is suggested that release of K+ by active neuronal compartments stimulates local protein synthesis of glial cells, resulting in the formation of glial sheaths with active K+ uptake capacity. Strong K+ release may even induce glial proliferation.

Animals↗
PubMed1989

Organelle-free cytoplasmic volume fraction of rabbit retinal Müller (glial) cells.

Retinal Müller (glial) cells are thought to act as "cables" carrying spatial buffering K+ currents from the sites of neuronal release into the reservoir of the vitrous body. In order to calculate the amplitude of such currents it is necessary to know the intracellular volume fraction which is able to carry these currents. Thus, this organelle-free volume fraction was measured in transmission electron microscopic photograms of rabbit Müller cells. This volume fraction was found to vary between 0.7 and more than 0.9 in various retinal layers except at the "external limiting membrane" where it was reduced to 0.24 by the accumulation of mitochondria. In enzymatically isolated cells all values are slightly increased by cell swelling.

Animals↗
PubMed1989

The course of axons within the postnatal rabbit retina.

The course of optic axons within the postnatal rabbit retina was studied by scanning electron microscopy (SEM) of the nerve fibre layer exposed by HCl and collagenase treatment, and by freeze-fracture electron microscopy. In the first days after birth, axon bundles were found to run along a rather tortuous way in between the developing Müller cell endfeet. Later on and in adults, the axon bundles show a more straight course. Single axons running into a bundle show a very irregular course before they assume their path in parallel to the other fibres of the bundle. These results support the idea that the course of growing axons is somehow determined by free "channels" in between the endfeet of glial cells.

Animals↗
PubMed1988

Cytotopographical specialization of enzymatically isolated rabbit retinal Müller (glial) cells: K+ conductivity of the cell membrane.

Müller (radial glial) cells were isolated from rabbit retinae by means of papaine and mechanical dissociation. Regional membrane properties of these cells were studied by intracellular microelectrode recordings of potential responses to local application of high K+ solutions. When different parts of the cell membrane were exposed to high K+, the amplitude of the depolarizing responses varied greatly, indicating a strong regional specialization of the membrane properties. Using morphometrical data of isolated rabbit Müller cells, and a simple circuit model, we calculated the endfoot membrane to constitute more than 80% of the total K+ conductance of the cell; the specific resistivity of the endfoot membrane was about 400 omega cm2, i.e., more than 40 times less than that of the membrane of the vitread process, which is immediately adjacent. This kind of regional membrane specialization seems to be optimized in respect to the Müller cells' ability to carry spatial buffering K+ currents.

Animals↗
PubMed1988

Efficient K+ buffering by mammalian retinal glial cells is due to cooperation of specialized ion channels.

Radial glial (Müller) cells were isolated from rabbit retinae by papaine and mechanical dissociation. Regional membrane properties of these cells were studied by using the patch-clamp technique. In the course of our experiments, we found three distinct types of large K+ conducting channels. The vitread process membrane was dominated by high conductance inwardly rectifying (HCR) channels which carried, in the open state, inward currents along a conductance of about 105 pS (symmetrical solutions with 140 mM K+) but almost no outward currents. In the membrane of the soma and the proximal distal process, we found low conductance inwardly rectifying (LCR) channels which had an open state-conductance of about 60 pS and showed rather weak rectification. The endfoot membrane, on the other hand, was found to contain non-rectifying very high conductance (VHC) channels with an open state-conductance of about 360 pS (same solutions). These results suggest that mammalian Müller cells express regional membrane specializations which are optimized to carry spatial buffering currents of excess K+ ions.

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