Biomedical subjects

A Reichenbach

Publications and source records attributed to A Reichenbach.

At least 91 records · Page 5Linked to original sources

PubMed1997

Retinal light damage vs. normal aging of rats: altered morphology, intermediate filament expression, and nuclear organization of Müller (glial) cells.

In retinal light damage, degeneration of photoreceptors results in alterations of glial (Müller) cells. In particular, Müller cells show signs of gliosis such as thickening of their stem processes, and expression of glial fibrillary acidic protein (GFAP) which is normally not detectable by immunocytochemistry. We were interested in a quantification of these morphological alterations, and in possible effects of an application of free radical scavengers (Ginkgobiloba extract EGb 761). For this purpose, we studied Müller cells in retinae of albino rats exposed to enhanced illumination for 24 months, a procedure which causes a complete loss of photoreceptor cells. The cells were labeled by (i) bulk filling with the fluorescent dye, Procion yellow, and by (ii) immunocytochemical demonstration of vimentin and GFAP. One group of rats was fed daily with EGb 761 during the last 8 months of life when the remaining photoreceptors (about 50%) died. The retinae were compared with retinae from 3 months-old albino rats, serving as normal young controls, and with retinae from 24 month-old pigment rats, representing normal aging processes. As age-related changes of the ultrastructure of glial cell (astrocytic) nuclei have been described in the literature, the organization of Müller cell nuclei was also studied by an argyrophilic stain, and by electron microscopy. We found that in the thin light-damaged retinae, Müller cells were shorter but thicker than in age-matched control retinae. The volumes of their vitread stem processes were almost unchanged. Müller cells were GFAP-immunoreactive in the light-damaged retinae but not in the controls. The application of EGb 761 prevented the expression by Müller cells of (detectable levels of) GFAP. By contrast, in retinae from EGb 761-treated animals the volumes of the vitread stem processes were significantly increased in comparison to untreated animals. The number of nuclear organization regions was significantly enhanced in Müller cell nuclei from light-damaged untreated albino rats, as compared with the young controls. Application of EGb 761 prevented much of this increase. Thus, exogeneous free radical scavengers do not prevent the occurrence of an reactive hypertrophy but inhibit the expression of "pathological marker molecules", and the (accompanying) signs of enhanced nuclear activity.

Aging↗

PubMed1996

Immunocytochemical demonstration of alpha 2-M-R/LRP on Müller (glial) cells isolated from rabbit and human retina.

The alpha 2-macroglobulin receptor/low-density lipoprotein receptor-related protein (alpha 2-M-R/LRP) is a multifunctional receptor which has been implicated in lipoprotein metabolism, clearance of proteinase-proteinase inhibitor complexes and regulation of growth factor/cytokine metabolism. This receptor is abundantly present in numerous tissues and organs such as liver, lung, placenta and brain. In brain it is expressed in neurones but not in normal macroglia. Using immunocytochemistry and monoclonal antibodies against the large extracellular receptor subunit we have detected alpha 2-M-R/LRP on enzymatically isolated retinal Müller (glial) cells. This receptor may be involved in vital functions of Müller cells.

Animals↗

PubMed1996

Ammonia-induced taurine release from cultured rabbit Müller cells is an osmoresistant process mediated by intracellular accumulation of cyclic AMP.

A previous study demonstrated the release of newly loaded radiolabelled taurine (Tau) from cultured rabbit Müller glia not only following typical cell volume-increasing treatments with high (65 mM) potassium ions or hypotonic media, but also with ammonium chloride (further referred to as ammonia), in a dose-dependent manner, at doses ranging from physiological (0.25 mM) to those accompanying hyperammonemic coma (5 mM) (Faff-Michalak et al., Glia 10:114-120, 1994). Stimulation of Tau release by ammonia, but not by 65 mM potassium, was correlated with a dose-dependent increase of intracellular cAMP levels. The release, as measured at 5 mM ammonia, was abolished by compounds that prevented cAMP increase: an adenylate cyclase inhibitor, miconazole, a protein kinase A inhibitor HA 1004, an anion channel blocker, niflumic acid, and a Tau transport site agonist, beta-alanine. The release by ammonia differed from potassium-induced release in its resistance to 1) increase of medium tonicity by addition of 50 mM sucrose; 2) addition of the anion/cation cotransport blocker, furosemide; and 3) removal of calcium from the superfusion medium. The results suggest that ammonia-induced Tau release is mediated by intracellular accumulation of cAMP and may occur either via an osmoresistant, cAMP-controlled channel or a cAMP-activated Tau transporter. The release observed at the physiological concentration of ammonium chloride suggest a role for ammonia as a signal molecule.

Ammonia↗

PubMed1996

The Müller cell: a functional element of the retina.

Müller cells are the principal glial cells of the retina, assuming many of the functions carried out by astrocytes, oligodendrocytes and ependymal cells in other CNS regions. Müller cells express numerous voltage-gated channels and neurotransmitter receptors, which recognize a variety of neuronal signals and trigger cell depolarization and intracellular Ca2+ waves. In turn, Müller cells modulate neuronal activity by regulating the extracellular concentration of neuroactive substances, including: (1) K+, which is transported via Müller-cell spatial-buffering currents; (2) glutamate and GABA, which are taken up by Müller-cell high-affinity carriers; and (3) H+, which is controlled by the action of Müller-cell Na(+)-HCO3- co-transport and carbonic anhydrase. The two-way communication between Müller cells and retinal neurons indicates that Müller cells play an active role in retinal function.

Animals↗

PubMed1995

Müller glial cells of the tree shrew retina.

The tree shrew is one of the few mammalian species whose retinae are strongly cone dominated, which is usually the case in reptilian and avian retinae. Müller cells of the tree shrew (Tupaia belangeri) retina were studied by transmission electron microscopy of tissue sections and freeze-fracture replicas, by immunolabeling of the intermediate filament protein vimentin in radial paraffin sections and in whole retinae, as well as by intracellular dye injection in slices of retinae. In addition, enzymatically isolated cells were stained by Pappenheim's panoptic staining method. The cells showed an ultrastructure that is similar to other mammalian Müller cells with two exceptions: Due to the extensive lateral fins of cone inner segments, the apical microvilli of Müller cells are arranged in peculiar palisades, and the basket-like Müller cell sheaths around neuronal somata in both nuclear layers consist of unusual multilayered membrane lamellae. Unlike Müller cells in other mammalian species studied thus far, but similar to reptilian and avian Müller cells, those of tree shrews commonly have two or more vitread processes rather than one main trunk. Müller cell densities range between some 13,000 mm-2 in the periphery and about 20,000 mm-2 in the retinal center. Neuron:(Müller)glial cell ratios were estimated to be 7.9:1 in the center and 6.2:1 in the periphery. For each Müller cell, about 1.5 (cone) photoreceptor cells, four or five interneurons of the inner nuclear layer, and about one cell of the ganglion cell layer were counted. This is a much lower number of neurons per Müller cell than in most other mammals studied.

Animals↗

PubMed1995

Development of A-type (axonless) horizontal cells in the rabbit retina.

The development of A-type horizontal cells (HC) was studied in the rabbit retina between embryonic day (E)24 and adulthood [the day of birth was called postnatal day (P)1 and corresponds to E31-32]. The cells were visualized by several methods 1) by immunolabeling with antibodies to neurofilament 70,000 (NF-70kD), 2) by immunolabeling with antibodies to a calcium binding protein (CaBP-28kD), 3) by two different methods of silver impregnation, and 4) by histochemical demonstration of NADH-diaphorase activity. Most methods labeled A-type HC only in the dorsal retina; thus, our study is restricted to HC of this region. HC densities were determined at each developmental stage. The cells were drawn at scale, and size, quotient of symmetry, and topographical orientation of dendritic trees were studied by image analysis. The growth of HC dendritic fields was correlated with data on the postnatal local retinal expansion, which is known to be driven by the intraocular pressure (after cessation of retinal cell proliferation at P9). This expansion was evaluated in an earlier paper (Reichenbach et al. [1993] Vis. Neurosci. 10:479-498) by using local subpopulations of Müller cells as "markers" of distinct topographic regions of the retinae. After E24, when the final number of HC is established, we can discriminate three distinct developmental stages of A-type HC. During the first stage, between E24 and E27, the young cells are often vertically oriented and may extend their first short dendrites within (the primordia of) both plexiform layers. The irregular HC mosaic at E24 shows a significant difference to all other stages. The second stage begins after birth when the dendritic trees of the cells are already restricted to the outer plexiform layer. Between P3 and P9, their dendritic trees enlarge more than the surrounding retinal tissue expands, and the coverage factor almost doubles from 2.5 to 4.4. The third stage occurs after P9 when the growth rate of dendritic tree areas corresponds to that of the local retinal tissue expansion caused by "passive stretching" of the postmitotic tissue, and the coverage factor remains constant. This is compatible with the view that mature synaptic connections of A-type HC are mostly established after the first week of life and are then maintained.

Animals↗

PubMed1995

Mammalian Müller (glial) cells express functional D2 dopamine receptors.

Dopamine plays important functional roles in the vertebrate retina. Here we show that functional D2 dopamine receptors are present on mammalian retinal Müller (glial) cells. Using an antiserum directed to two oligopeptides predicted from rat D2 receptor DNA, patchy label was demonstrated immunocytochemically on virtually all Müller cells enzymatically isolated from guinea-pig and rat retinae. Application of exogeneous dopamine to voltage-clamped isolated living guinea-pig Müller cells caused either a decrease (40%), an increase (32%), or no change (28%) of the input resistance of the membrane. The D2 receptor agonist quinelorane caused an increase of the membrane's input resistance in 100% of the cells. This effect was completely blocked by the D2 receptor antagonist S(-)-sulpiride. When all voltage-activated K+ channels except the delayed rectifiers were blocked by Ba2+, quinelorane had no effect. Further, the reversal potentials of the responses were near the potassium equilibrium potential. We conclude that the activation of Müller cell D2 receptors closes (inwardly rectifying) K+ channels. The presence of functional dopamine receptors on mammalian Müller cells may have important consequences for retinal K+ clearance, and thus, for information processing in the retina.

Animals↗

PubMed1995

Expression of glial fibrillary acidic protein (GFAP), glutamine synthetase (GS), and Bcl-2 protooncogene protein by Müller (glial) cells in retinal light damage of rats.

In retinal light damage, degeneration of photoreceptors may cause alterations of glial (Müller) cells. We performed immunocytochemical studies on Müller cells isolated from retinae of rats exposed to enhanced illumination for 24 months, a procedure which leads to complete loss of photoreceptor cells. One group of rats was fed daily with Ginkgo biloba extract (EGb 761, an established free radical-scavenger) during the last 8 months of life when the remaining photoreceptors (about 50%) die. We found that (1) Müller cells respond to photoreceptor damage by increased expression of glial fibrillary acidic protein, (2) Müller cells reduce expression of glutamine synthetase when the major glutamate-releasing neurons are lost, and (3) the application of exogenous free radical scavengers prevents the expression by Müller cells of the protooncogene protein Bcl-2, a molecule assumed to activate endogenous free radical-scavenging activities.

Animals↗

PubMed1995

Effects of enhanced extracellular ammonia concentration on cultured mammalian retinal glial (Müller) cells.

Müller (glial) cells of the neonatal rabbit retina were cultured as confluent monolayers and exposed to enhanced concentrations of ammonia (0.25, 0.5, 1, 3, 7, and 10 mM) in medium for various periods (30 min to 10 d). This caused, in a time- and dose-dependent manner, similar changes in the Müller cells as had previously been described in cultured astrocytes. The most conspicuous events were 1) an increasing size of cell nuclei, 2) an accumulation of phagocytotic vacuoles, and 3) a rearrangement of intermediate filaments. 4) A considerable number of cells died when higher ammonia concentrations were applied for more than 1 h. Simultaneous application of dibutyryl-cyclic adenosine monophosphate (dBcAMP) prevented almost completely both the increase in cell nucleus size and the changes of intermediate filaments, but only partly the early cell death of a subpopulation of cells, and the accumulation of phagocytotic vacuoles. Further changes evoked by enhanced ammonia concentration were 5) an accumulation of lipofuscin-like material ("fatty degeneration") revealed by lipophilic stain, 6) reduced immunoreactivity for cathepsin D, and increased immunoreactivity for 7) glial fibrillary acidic protein, 8) glutamine synthetase, and 9) bcl-2 protooncogene protein. These findings are discussed in respect to the possible underlying pathophysiological mechanisms.

Ammonia↗

PubMed1995

Hepatic retinopathy: morphological features of retinal glial (Müller) cells accompanying hepatic failure.

More than 80 years ago, Alzheimer described changes in the brains of patients who had suffered hepatic failure. Astrocytes are primarily affected; their nuclei become swollen, their intermediate filament protein composition is altered and their cytoplasm becomes vacuolated. Cells with these features are called Alzheimer type II astrocytes and these changes have been attributed to the toxic effects of elevated ammonia levels. The present study investigates whether the dominant glia of another part of the central nervous system, the Müller cells of the retina, undergo similar changes. Retinae of patients who had died with symptoms of hepatic failure were processed for histology, histochemistry, and immunocytochemistry. Cell nuclei were measured from brain astrocytes (insula cortex), Müller cells, and retinal bipolar neurons. Hepatic failure resulted in the enlargement of nuclei in astrocytes and Müller cells, and the enhanced expression in Müller cells of glial fibrillary acidic protein, cathepsin D, and the beta-subunit of prolyl 4-hydroxylase (glial-p55). In some retinae, signs of gliosis were also observed. We conclude that increased levels of serum ammonia resulting from hepatic insufficiency cause changes in Müller cells that are similar to those seen in brain astrocytes. We term this condition hepatic retinopathy.

Adult↗

PubMed1995

Alterations of Müller (glial) cells in dystrophic retinae of RCS rats.

We have carried out a light microscopical study of Müller cells in the retinae of rats with inherited retinal dystrophy (Royal College of Surgeons rats). Isolated retinae of both control and Royal College of Surgeons rats were exposed to a Procion Yellow solution which is taken up selectively into Müller cells. The shape of the cells was then studied by confocal microscopy. Enzymatically isolated Müller cells were studied immunocytochemically with antibodies against glial fibrillary acidic protein, cathepsin D, beta-amyloid precursor protein, bcl-2 protooncogene product, and glutamine synthetase. Müller cells from RCS retinae were shorter than those from control retinae, and showed a coarse hypertrophy of their distal (sclerad) processes. In Müller cells isolated from the retinae of Royal College of Surgeon's rats, the expression of glial fibrillary acidic protein, cathepsin D, beta-amyloid precursor protein and bcl-2 protooncogene product was increased, and the expression of glutamine synthetase was reduced. Obviously, loss of neighbouring neurons leads to major alterations of both the shape and metabolism of Müller cells. The expression of enzymes that serve functional glio-neuronal interactions, such as glutamine synthetase, seems to be down-regulated, whereas proteins involved in cell reconstruction (cathepsin D), cell repair (possibly beta-amyloid precursor protein), and protection against apoptotic cell death (bcl-2 protooncogene product), are up-regulated, together with the 'pathological marker' glial fibrillary acidic protein.

Amyloid beta-Protein Precursor↗

PubMed1995

Visualization of Müller (retinal glial) cells by bulk filling with procion yellow.

A method is presented that allows for an easy and reliable demonstration of retinal glial (Müller) cell morphology. When a 3% solution of the fluorescent dye Procion Yellow (reactive yellow, Sigma) is placed on isolated living retinae for 2 hrs, many Müller cells take up the dye. In paraffin sections, the cells can be observed by confocal microscopy in great detail. As the cells are filled throughout their length, the method has advantages over most immunocytochemical methods which label only parts of the cells. The method was applied to retinae of frogs, rats, guinea pigs, and rabbits. The vitread trunks of the cells differed in diameter. Those of frogs and rats were thin (less than 1 to 2 microns diameter) whereas those of guinea pigs and rabbits were thicker (2 to 5 microns). In all species studied the following rule was found. In thick central regions of the retina, Müller cells were long with slender trunks whereas in the thin retinal periphery, the Müller cells had thick short trunks. There was an inverse relationship between length and diameter of Müller cell trunks. Mammalian Müller cells were densely packed and had rather cylindrical endfeet. In the frog retina, Müller cells were more sparsely distributed, and the endfeet formed wide, flat funnels. It is concluded that the higher metabolic rate of mammalian retinae requires more densely packed Müller cells than occur in the amphibian retina.

Animals↗

PubMed1995

Distribution of Bergmann glial somata and processes: implications for function.

We have used immunocytochemistry for glial fibrillary adidic protein and glutamine synthetase to selectively label Bergmann glia in the adult rat cerebellum. From measurements of radial, tangential and en face sections we provide new data on the distribution and disposition of these glial cells. Specifically, Bergmann glia were found to have a mean areal packing density of 8,269 somata/mm2, their radial processes are packed at a mean density of 39,000/mm2, and their endfeet at the pial surface have a mean density of 19,973/mm2. Each Bergmann glial cell ist "responsible" for the equivalent of a column of cerebellar cortex having a base of 11 microns x 11 microns, a height of 170 microns, and a volume of 20,559 microns3. There are 8.1 Bergmann glia for each Purkinje cell, and each glial cell ensheaths between 2,142 and 6,358 Purkinje cell synapses. We use these data to offer insights on the roles of Bergmann glia during development and in the adult brain.

Animals↗

PubMed1995

Surface complexity of human neocortical astrocytic cells: changes with development, aging, and dementia.

Astrocytes from area 11 of the human neocortex were Golgi-impregnated. The material was taken from four human subjects, (1) a neonate without obvious CNS damage, (2) a middle-aged adult serving as control, (3) an aged individual without apparent mental disturbances, and (4) an aged patient with dementia. Camera-lucida drawings of 7 to 26 astrocytic cells from each donor were digitized into a personal computer, and analyzed for their fractal dimension D which is an objective and quantitative measure of the complexity of the cell's borders. Both radial and multipolar astrocytes from neonatal cortex showed a comparatively low fractal dimension (D approximately 1.39), that is, low surface complexity. Adult ("control") astrocytes were characterized by a high fractal dimension (D = 1.57), corresponding to their large surface complexity. Astrocytes from the aged subject showed a somewhat smaller fractal dimension (D = 1.51) and, thus, surface complexity; this difference was however not highly significant. In contrast, astrocytes from the subject with dementia showed a significant reduction in surface complexity (D = 1.47). Parallel changes were observed in the brain volume compartment occupied by the process of individual astrocytes. These data suggest that formation and maintenance of astrocytic processes and/or side branches correlate positively with the degree of information processing within their neuronal environment.

Adult↗

PubMed1994

Development of the rabbit retina. V. The question of 'columnar units'.

A qualitative and quantitative description of the columnar units in the mammalian retina, and a discussion of their ontogeny and putative functions is given. Columnar arrangements of cells exist in the developing retina which can be observed by means of scanning electron microscopy. In the adult retina, each Müller cell ensheaths a columnar group of neuronal cells. Counting the number of cells in radial H/E stained sections at various developmental stages reveals a constant ratio of neuronal cells per Müller cell, independent of the developmental stage (after postnatal day 9), and independent of the retinal topography. Such groups of cells always consist of one Müller cell, 11 rod photoreceptor cells, about 2 bipolar cells, and 1 to 2 amacrine cells. Retinal ganglion cells, cone photoreceptor cells, and horizontal cells are more sparsely distributed in the retina than these units; since they are known to arise earlier in the ontogenesis than other cell types they are considered to exist independently of the columnar units. It is suggested that the units arise by migration of groups of preneurons along a common Müller (precursor) cell; these preneurons and the corresponding Müller cell may be clonally related. In the adult retina, such columns might constitute metabolic and functional units.

Animals↗

PubMed1994

Immunocytochemical demonstration of astrocytes and microglia in the whale brain.

Whale brains have attracted the attention of neuroscientists but there are only sparse studies on whale glial cells. Here we report on immunolabeling of astrocytes by antibodies to glial fibrillary acidic protein (GFAP) or protein S-100 beta (both by the streptavidin/biotin technique), and labeling of microglial cells by Griffonia simplicifolia agglutinin (GSA I-B4, coupled to horseradish peroxidase), in the neocortex of a harbour porpoise Phocoena phocoena L. Many subpial and perivascular astrocytes were stained; they differed greatly in thickness and length of their processes. Subpial astrocytes were coarse with a few stout stem processes, whereas perivascular astrocytes deeper in the brain had many long and slender processes. Additionally, some long radial astrocytes were observed. Microglia were labeled throughout the brain, and showed similar features as 'resting' (ramified) microglia in the brain of other mammals.

Animals↗

PubMed1994

K(+)-, hypoosmolarity-, and NH4(+)-induced taurine release from cultured rabbit Müller cells: role of Na+ and Cl- ions and relation to cell volume changes.

The release of preloaded radiolabeled taurine (TAU) from cultured rabbit Müller cells [14-21 days in vitro (DIV)] was measured before and after treatment with the following stimuli: 1) isoosmotic 65 mM KCl; 2) a medium made hypoosmotic by uncompensated lowering of Na+ by 40-100 mM; and 3) NH4Cl ranging from 0.25 to 5 mM. The same stimuli were tested for their effect on the cell volume by the 3-O-methyl-D-glucose (OMG) uptake method of Kletzien et al. (Anal Biochem 68:537, 1975). Hypoosmotic media and 65 mM KCl stimulated TAU release, and the release was well correlated with the increase of cell volume. The stimulatory effect of 65 mM KCl was abolished by isotonic removal of Cl- or Na+, and omission of either ion markedly enhanced the basal release of TAU. The results are roughly consistent with the characteristics of the swelling-induced TAU release reported for cultured astrocytes and neurons of various CNS regions, and also for freshly isolated, nondissociated retina. Taken together, the results are indicative of a significant role of TAU release from Müller cells, in the osmosensory response of the retina. Ammonium chloride stimulated TAU release in a dose-dependent manner, a significant stimulation being already observed at 0.5 mM, a concentration that is frequently measured in brain during acute hyperammonemia. The effect of NH4Cl was strictly chloride dependent at 0.5-2 mM, but partly Cl- independent at 5 mM. The Kletzien's method did not appear to be well suited for measuring cell volume in the presence of ammonium ions.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Na+ channels of Müller (glial) cells isolated from retinae of various mammalian species including man.

Within the last few years, the expression of voltage-dependent, TTX-sensitive Na+ channels has been demonstrated in several types of neuroglial cells such as astrocytes and Schwann cells. Recently, we reported the occurrence of such Na+ currents in retinal Müller (glial) cells from dog and cat. This paper deals with the description of the properties of Na+ currents in Müller cells isolated from retinae of several mammalian species, as well as from human retinae. These Na+ currents were eliminated by TTX (1 microM), and by exposure to sodium-free extracellular solution; typically, they were demonstrable only after blocking most of the K+ conductance by Ba2+ (1 mM). Voltage-dependent activation and inactivation characteristics and time constants of the Na+ currents were similar to those of currents carried by neuronal Na+ channels. The estimated number of sodium channels per cell was low (about 1,500 channels per 7,500 microns 2), and the K+ conductance exceeded the peak Na+ conductance by an average factor of 5. Thus, the cells were incapable of generating action-potential-like responses under current clamp. Modelling estimations show that triggering of glial Na+ currents under physiological conditions, if any, can at best occur by emhaptic transmission at perinodal sites of optic axons. It is speculated that glial Na+ channels might be involved in neuroglial signalling events.

Animals↗