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Biomedical subjects

R Dermietzel

Publications and source records attributed to R Dermietzel.

At least 55 records · Page 3Linked to original sources

Immunolocalization of the neural cell adhesion molecule L1 in non-proliferating epithelial cells of the male urogenital tract.

The localization of the neural cell adhesion molecule L1 in the male urogenital tract (including seminal vesicles and prostate) of the mouse and bull was investigated using immunocytochemical and immunochemical methods in order to better understand the function of this glycoprotein in non-neural tissues. L1 antibodies labeled non-myelinated nerves in all portions of the urogenital tract investigated. However, L1 immunoreactivity was also found between epithelial cells of several regions of the urogenital system including epididymal tail, deferent duct, ejaculatory duct and seminal vesicles. Some L1 immunoreactivity was also demonstrated between epithelial cells of murine urinary bladder and urethra. The specificity of the immunoreaction was verified by western blots. There was no correlation between L1 expression and proliferating activity as revealed by double immunocytochemistry using various markers of cell proliferation. This unexpected expression of L1 in nonneural tissues is mainly restricted to non-proliferating epithelia of those portions of the urogenital tract that are derived from the Wolffian duct. It is suggested that L1 in these epithelia could enhance the mechanical resistance and reduce transepithelial permeability.

Animals↗

Changes in the expression pattern of blood-brain barrier-associated pericytic aminopeptidase N (pAP N) in the course of acute experimental autoimmune encephalomyelitis.

The role of cerebral pericytes in blood-brain barrier (BBB) mechanisms is still a matter of controversy. Because acute experimental autoimmune encephalomyelitis (EAE) is characterized by a transient and focal perturbation of the BBB, we have utilized the model of adoptive transfer EAE to correlate the expression of the pericytic aminopeptidase N (pAP N) with the acute functional state of the BBB. We demonstrate that a significant downregulation of microvascular pAP N expression occurs, and the observed perturbation of the enzymatic BBB complement seems to be a sustained effect which persists even after recovery from clinical disease. At the peak of clinical disease, numerous pAP N expressing invasive cells were detected in white matter of the lumbar spinal cord. Through the use of a panel of different immunocytochemical markers these pAP N-immunopositive cells were characterized as ED 1-positive, most likely hematogenous macrophages. Activated resident microglial cells participate in the EAE-induced inflammatory process to only a minor extent.

Acute Disease↗

X-linked dominant Charcot-Marie-Tooth disease and other potential gap-junction diseases of the nervous system.

Gap junctions play important roles in the exchange of information and metabolites in the nervous system. These roles are highlighted by peripheral neuropathy (X-linked dominant Charcot-Marie-Tooth disease) that is associated with mutations in a gap-junction protein (connexin32), resulting in loss of function, and by somatic dysfunctions where changes in expression, organization or function of gap junctions are associated with neuronal hyper- or hypoexcitability. In this review, the causes and consequences of this gap-junction-related peripheral neuropathy and other pathological conditions of the nervous system, where dysfunctions of junctional communication are considered to play a casual role, are considered.

Charcot-Marie-Tooth Disease↗

Expression of the tight junction protein ZO-1 in the olfactory system: presence of ZO-1 on olfactory sensory neurons and glial cells.

The olfactory system is a unique part of the central nervous system since it retains neuronal turnover and regenerative capacities in adulthood. Thus it provides an ideal model to study plasticity of membrane moities involved in cell-cell interactions. One structure particularly involved in cell-cell interaction is the tight junction, which establishes polarization of epithelial cells and creates diffusion barriers to paracellular passages. ZO-1 is a phosphoprotein peripherally associated with tight junctions. We have studied expression of ZO-1 protein in the developing and adult olfactory system of the mouse in order to get information about the localization and developmental expression of this tight junction component. ZO-1 expression has also been determined in cell cultures of olfactory bulbs. ZO-1 was present in the olfactory placode prior to formation of tight junctions. ZO-1 was localized in the developing and mature olfactory epithelium at heterotypic contacts between supporting cells and olfactory neurons as well as at homotypic contacts between both these cell types. Confocal microscopy showed quantitative differences in the ZO-1 expression among different olfactory dendrites. In the olfactory nerves ZO-1 immunolabeling was detectable between olfactory ensheathing cells. From the seventh postnatal day ZO-1 immunolabeling was detected at the mitral cell layer of the bulb on cells tentatively identified as oligodendrocytes. Myelinated tracts of the bulb were ZO-1 negative. Cell cultures of olfactory bulbs showed ZO-1 immunoreaction, mostly localized on glial fibrillary acidic protein (GFAP)-positive cells. Our results provide further evidence that ZO-1 serves functions unrelated to the tight junction complex and indicate molecular heterogeneity of these cell-cell contacts.

Animals↗

Cloning and in situ localization of a brain-derived porin that constitutes a large-conductance anion channel in astrocytic plasma membranes.

We have cloned a protein from bovine brain, brain-derived voltage-dependent anion channel 1 (BR1-VDAC), that is identical to a recently sequenced plasmalemmal-bound porin from human lymphocytes. mRNA hybridization indicates that BR1-VDAC is widely distributed throughout nervous and nonnervous tissues. In situ localization substantiated that the BR1-VDAC is associated with the plasmalemma of astrocytes. A monoclonal antibody that recognizes the N terminus of the BR1-VDAC protein completely blocks an astrocytic high-conductance anion channel that has electrophysiological similarities with the mitochondrial VDAC. Since the high-conductance anion channel in astrocytes has been shown to respond to hypoosmotic solutions, its molecular identification provides the basis for a better understanding of volume regulation in brain tissue.

Animals↗

Astrocytes as rapid sensors of peripheral axotomy in the facial nucleus of rats.

Facial nerve transection leads to functional and structural reactions in lesioned motor neurones and surrounding glial cells. Data from this study provide evidence that the most rapid reaction described so far consists of an increase in immunoreactivity of connexin-43 (cx-43), the predominant gap junction protein in astrocytes. The ipsilateral facial nucleus is selectively marked as early as 0.75 to 1.5 hours after axotomy, while the unlesioned side as well as the unoperated controls remain faintly stained. Thus, enhanced coupling capacity of astrocytes by gap junctions appears to be a sensitive indicator of modified neuronal-glial interaction in the CNS.

Animals↗

The 140-kDa protein of blood-brain barrier-associated pericytes is identical to aminopeptidase N.

Although ample evidence has been accumulated on the structure and functional role of endothelial cells in blood-brain barrier mechanisms, little is known about the contribution that cerebral pericytes provide to this phenomenon. We have reported recently on a monoclonal antibody specific for cerebral pericytes at blood-brain barrier sites. To confirm the pericytic localization of this antigen, and in order to elucidate its biochemical identity, we have performed immunocytochemical, biochemical, and molecular biological studies. By immunocytochemistry on the light microscopic as well as electron microscopic level, we provide definite evidence that the 140-kDa antigen recognized by this monoclonal antibody is confined to cerebral pericytes, whereas endothelial cells are devoid of this antigen. N-Terminal sequencing of the corresponding immunocrossreacting renal protein revealed that the protein detected by the monoclonal antibody is identical to aminopeptidase N. By means of the reverse transcriptase-polymerase chain reaction, the identity of the 140-kDa antigen as aminopeptidase N could also be verified for cerebral microvascular cells. Cerebral pericytic aminopeptidase N may be involved in neurotransmitter (enkephalin) metabolism at the blood-brain interface. By taking into account that brain pericytes have been found to express further plasma membrane-bound enzymes, these results strongly suggest the contribution of cerebral pericytes in the metabolic concert of the homeostatic balance regulated by the blood-brain barrier.

Amino Acid Sequence↗

Maturation of renal collecting duct cells in vivo and under perifusion culture.

The embryonic collecting duct epithelium of neonatal kidney undergoes profound functional changes during maturation. In its initial state as inductor epithelium it appears homogeneous, but differentiates into a heterogeneously composed collecting duct epithelium consisting of principal and intercalated cells. The mechanism of this terminal differentiation process is unknown. We used morphological and immunohistochemical methods to investigate the maturation of the collecting duct system in neonatal rabbit kidney and under organotypic culture conditions. The new perifusion culturing method allowed us to follow the differentiation of the ampullary collecting duct epithelium under conditions as close as possible to the situation within the organ. With this technique we were able to induce a differentiation process similar to that in the in situ situation. This process led to the appearance of a mixed cell population consisting of principal and intercalated-like cells, respectively. A continuous perifusion of the medium made it possible to stabilize the microenvironment under culture conditions and thus to maintain the heterogeneous composed collecting duct epithelium in a differentiated status over long periods of time.

Animals↗

Synapses between slowly adapting lung stretch receptor afferents and inspiratory beta-neurons in the nucleus of the solitary tract of cats: a light and electron microscopic analysis.

Previous neuroanatomic and physiologic studies indicated that afferent fibres from slowly adapting pulmonary stretch receptors (SAR) project to the nuclei of the solitary tract and terminate on inspiratory beta-neurons. In the present study we combined electrophysiologic and morphologic approaches to verify the presumed monosynaptic connections between SARs and beta-neurons. Single identified beta-neurons and single identified SAR afferent fibres were labelled intrasomally and intraaxonally, respectively, with horseradish peroxidase (HRP) in the same anesthetized cats. Under the light microscope, we analyzed the morphology of beta-neurons and their dendritic fields and of the terminal projection pattern of fibres from SARs and identified potential synaptic connections between boutons of SAR afferent fibres and the soma and dendrites of beta-neurons. The identified tissue was then processed further for electron microscopic analysis. On average, beta-neurons had 6 primary dendrites that bifurcated 3-8 times. The dendritic trees extended 1.5 mm both rostrocaudally in the ventrolateral nucleus of the solitary tract and medially into the intermediate subnucleus. Axons of beta-neurons curved toward the midline and no collateral branches were evident over its stained length (2.5-3.4 mm). Axodendritic synaptic contacts between SAR fibres and beta-neurons were identified electron microscopically in four of six tissue samples chosen by light microscopy. In addition, we located 2 axodendritic and 2 axosomatic synaptic contacts that were not observed under light microscopic screening. The boutons of SAR fibres contained clear, round vesicles and formed asymmetrical synapses with beta-neurons. Multiple synaptic connections were found between collaterals of a single SAR and single beta-neurons, indicating a dense terminal projection of single SAR afferent fibres onto beta-neurons. These morphologic data prove monosynaptic connections between electrophysiologically identified SAR afferent fibres and beta-neurons.

Animals↗

Facial nerve lesions lead to increased immunostaining of the astrocytic gap junction protein (connexin 43) in the corresponding facial nucleus of rats.

After peripheral transection of the facial nerve, immunostaining of astrocytic gap junction protein changed in the corresponding brainstem nucleus of the rat. Enhanced connexin-43 immunoreactivity was restricted to the ipsilateral facial nucleus and to astrocytes surrounding lesioned motoneurons. This reaction is focally distinct, and marks only a part of the astrocytic network indicating a local plasticity of intercellular coupling. These results suggest that astrocytes work as sensors of signals which either depend on the integrity of neighboring neurons or inform about neuronal disorders.

Animals↗

Characterization of bovine ciliary pigmented epithelial cells in monolayer culture: an ultrastructural, enzyme histochemical and immunohistochemical study.

Monolayer cultures of the pigmented epithelial (PE) cells derived from two regions of the pars plicata of bovine eyes were established and grown up to the third passage. After this passage, the cultures became senescent. During the first three passages, the PE cells lost their pigment granules but developed a distinct cellular polarity by forming junctional complexes at their apical cell portions and depositing basement membrane like material on their basal side. The junctional complexes were shown to be impermeable for horseradish peroxidase, suggesting that they contained tight junctions. Histochemically, the monolayer cells stained for carbonic anhydrase (CA) and Na+/K(+)-ATPase, enzymes involved in active fluid secretion. Staining for CA and Na+/K(+)-ATPase as well as for acid phosphatase and immunostaining for vimentin and actin of the cultured PE cells were comparable with that of PE cells in vivo. Therefore, PE monolayer cultures are considered to be a suitable model for experimental studies in vitro.

Acid Phosphatase↗

Gap junctions in the brain: where, what type, how many and why?

Gap junctions represent well-documented means of intercellular communication in various tissues, including the brain, where they function as portals allowing the exchange of electrolytes, second messengers and metabolites between cells. In view of the enormous recent surge of information dealing with the cellular and molecular biology of gap junctions in non-nervous tissue, as well as current interest in the cell biology of glia, this review is intended to provide an overview of the molecular and functional implications of gap-junction-mediated intercellular communication in the nervous system.

Animals↗

Aldosterone modulates PNA binding cell isoforms within renal collecting duct epithelium.

To investigate the differentiation of the ampullary collecting duct cells into adult principal and intercalated cells, the embryonic cortex of newborn New Zealand rabbit kidney was isolated and brought in culture. With this culture technique the ampullary cells formed a polarized collecting duct epithelium which was kept under permanent exchange of medium and in the presence of aldosterone, arginine vasopressin and/or insulin. After 14 days of perfusion culture the epithelia showed light and dark cells resembling the principal and intercalated cells of the adult collecting duct. The differentiation from embryonic into adult collecting duct cells was controlled by applying the monoclonal antibody CD 7. Independent of the hormonal treatment all of the epithelial cells matured in culture and expressed the CD 7 antigen. This corresponded with the situation found within the adult kidney, where the CD 7 antigen was localized in all principal and intercalated (IC) cells, whereas the embryonic ampullary epithelium in the neonatal kidney remained negative. A differentiation feature of the beta-type intercalated cell was investigated by labeling the cultured epithelia with peanut agglutinin (PNA). In contrast to the CD 7 antigen the development of PNA binding was highly dependent of time and individual hormone administration. While in control epithelia only 8% of PNA positive cells were found, aldosterone induced epithelia revealed 72% PNA labeled cells. The combination of aldosterone and insulin increased the number of PNA-positive cells to 90%. By scanning electron microscopy it could further be shown that several isoforms of cells were reactive with PNA. Thus, in culture the PNA label is not restricted to the typical beta-type IC cells.

Aldosterone↗

Expression of connexins in the developing olfactory system of the mouse.

To gain insight into the function of gap junctions' connexin43, connexin32 and connexin26 in a neural structure that retains neuronal turnover capacities throughout adulthood, the expression of these molecules has been investigated in the developing and adult olfactory system by immunocytochemical and biochemical methods. Connexin43 was detectable from the olfactory placode stage. During early embryonic development, the levels of connexin43 expression remained low. An increase in the expression of this connexin occurred perinatally. Expression of connexin43 became very high during the postnatal stages and adulthood. Electron microscopy (EM) immunocytochemistry of the olfactory system showed connexin43 expression in non-neuronal cells. Strong regional differences in the expression of connexin43 in the olfactory epithelium were observed. No apparent relationship between connexin43 expression and turnover activity of olfactory neurons was detected. Western blots of olfactory tissues revealed the presence of three different isoforms of connexin43. Connexin32 was detected in the olfactory bulb at late postnatal stages including adulthood. Connexin32 was observed on some cells tentatively identified as oligodendrocytes. Connexin26 was localized onto leptomeninges. Some immunofluorescence was also obtained in the periglomerular region and in the subependymal layer of the bulb. Northern blot analysis revealed the presence of mRNA of connexin32 and connexin26 in the adult olfactory system. Our results substantiate the cell specific expression of these three types of connexins and they document the primary of connexin43 in olfactory tissues. Moreover, our findings indicate that although expression of connexin43 in the olfactory system is developmentally regulated, it is not directly associated with the neuronal cell turnover of the olfactory epithelium.

Animals↗

Immunocytochemical localization of cell adhesion molecules in the developing and mature olfactory system.

The localization of Ca+(+)-independent cell adhesion molecules (CAMs) in the developing and mature olfactory epithelium and bulb is reviewed. The CAMs included in this article are the neural cell adhesion molecule (N-CAM), the 180 kD component of N-CAM (N-CAM 180), the embryonic form of N-CAM (E-N-CAM), L1 glycoproteins, J1 glycoproteins, and the adhesion molecule on glia (AMOG). In addition, the expression of the L2-HNK-1 carbohydrate epitope, shared by N-CAM, L1, J1 and myelin-associated glycoprotein (MAG) in the adult olfactory epithelium and bulb has also been documented. For the localization of these molecules at the light and electron microscopic levels, immunocytochemical techniques were used and are described in detail. During development and organogenesis, the olfactory system exhibits a pattern of CAM expression similar to the general pattern described for the developing nervous system. In the adult olfactory system, however, a significant retention of CAMs characteristic for developmental and morphogenetic processes, such as E-N-CAM, AMOG, as well as the high molecular weight components of J1 glycoproteins, can be observed. The retention of these embryonic features are most likely associated with the cell turnover and high plasticity of this system. Moreover, the predominance of N-CAM 180 with respect to other components of N-CAM, as well as the absence of the L2/HNK-1 carbohydrate epitope, are also particular traits of the primary olfactory system which could be associated with its exceptional properties.

Adenosine Triphosphatases↗