Sialic acid residues indirectly modulate the binding properties of AMPA-type glutamate receptors

Manipulations that disrupt the extracellular interactions of neural cell adhesion molecules (NCAMs) block the formation of stable long-term potentiation (LTP) but do not reverse already established potentiation. Several studies have implicated a change in AMPA-type glutamate receptors as being responsible for the expression of LTP but there are no evident links between NCAMs and the receptors. NCAMs are major carriers of sialic acid residues in the brain and removal of these with neuraminidase markedly affects the binding properties of the adhesion molecules. Therefore, the present study tested if neuraminidase treatment produces a change in AMPA receptors. Preincubation of cortical membranes with the enzyme for 15 min at 37°C caused a 5% reduction in the apparent sizes of NCAMs 140 and 180 but had no detectable influence on the sizes of various glutamate receptor subunits. The same treatment resulted in a 20±1% increase in the binding of [³H]AMPA with no apparent effect on binding to NMDA-type glutamate receptors or to high affinity kainate receptors. In membranes from the hippocampus, neuraminidase induced a 30±2% increase in binding which Scatchard analyses showed to be due to an increase in receptor affinity. Finally, neuraminidase had no effect on either the binding properties of solubilized AMPA receptors or on AMPA receptors stably expressed in a non-neuronal cell line. These results: (i) demonstrate that modulation of the extracellular environment can influence the binding properties of AMPA receptors, (ii) indicate that sialic acid residues in the extracellular compartment of synapses exert a significant and indirect influence on AMPA receptors and, (iii) suggest a route whereby NCAMs and LTP could be linked.     

NCAM as a differentiation marker of postmigratory immature neurons in the developing human nervous system

The regional distribution and cellular localization of the neural cell adhesion molecule were investigated in the developing nervous system of human embryos and fetuses, by using immunocytochemistry with antibodies against the human neural cell adhesion molecule. Embryos and fetuses with crown-rump lengths ranging from 15 to 80 mm (corresponding approximately to 6–12 ovulation weeks) were examined. In the telencephalon, immunoreactivity was found in the primordial plexiform layer and in the early cortical plate. During later stages, immunoreactivity was present in the marginal zone, cortical plate, developing subplate zone and radiations of the internal capsule. In the mesencephalon and rhombencephalon, neuronal perikarya were outlined and strong staining was present in fiber bundles. In the spinal cord, strong immunostaining was present in fiber tracts and in dorsal and ventral roots. Immunostaining outlined perikarya of dorsal and ventral gray columns; the neuropil of the developing gray matter was also stained. Contrary to findings in some other species, precursor cells in the proliferating zones, i.e. all cells of ventricular zones and the external granular layer in the developing cerebellum, were not stained. Reactivity, however, was also observed in single mature-looking nerve cell bodies and their processes, which were localized in proliferating zones. All peripheral nervous structures including nerve fibers and peripheral nervous tissue components derived from the neural crest (i.e. sensory and autonomic ganglia) expressed the neural cell adhesion molecule. At the cellular level staining always appeared to be associated with the entire cell membrane of a given neuron including its outgrowing processes. Immunocytochemical staining for neural cell adhesion molecule is an excellent method for the identification of single nerve fibers, nerves and nerve cell bodies in situ in the developing peripheral and early central nervous system. The neural cell adhesion molecule is suggested as a marker of postmigratory immature neurons in the developing human nervous system.     

CD56 (NCAM)-Positive Malignant Lymphoma

CD56, a natural killer cell marker reactive with the neuronal-cell adhesion molecule (NCAM), identifies a group of lymphomas with distinctive clinicopathologic features. The disease affects mostly middle-aged adults who often present with fever, skin rash and hepatospleno-megaly in the absence of peripheral lymphadenopathy. Extranodal involvement is common, particularly the skin, aerodigestive tract and central nervous system. Histologically, an an-giocentric and angiodestructive pattern of infiltrate is often seen, but the cytological spectrum of the lymphoma cells is very broad. Cytoplasmic granules, however, are frequently found when Giemsa-stained cytologic preparations are examined. Immunologically, CD56-positive lymphomas can be sub-classified into CD3-positive (T-cell) and CD3-negative (probably true natural killer cell) subtypes. T-cell receptor gene rearrangement can be demonstrated in the former cases, but not in the latter. Clinically, CD56-positive lymphomas are aggressive neoplasms.     

The neural cell adhesion molecule (NCAM) in development and plasticity of the nervous system

—The neural cell adhesion molecule (NCAM) is a member of the immunoglobulin superfamily and is strongly expressed in the nervous system. NCAM is found in three major forms, of which two—NCAM-140 and NCAM-180—are transmembrane proteins, while the third—NCAM-120—is attached to the membrane via a glycosylphosphatidyl inositol anchor. In addition, soluble NCAM forms exist in brain, cerebrospinal fluid, and plasma. NCAM mediates cell adhesion through homophilic as well as through heterophilic interactions. Following NCAM binding, transmembrane signalling is believed to be activated, resulting in increased intracellular calcium. By mediating cell adhesion to other cells and to the extracellular matrix and by activating intracellular signaling pathways, NCAM influences cell migration, neurite extension, and fasciculation, and possibly formation of synapses in the brain. From studies on NCAM knock-out mice, NCAM have been shown to be crucial for the formation of the olfactory bulb and the mossy fiber system in the hippocampus. In addition, NCAM is important for neuronal plasticity in the adult brain associated with learning and regeneration.     

Different expression of adhesion molecules on stromal cells and endothelial cells of capillary hemangioblastoma

Cell-cell and cell-matrix interactions are essential for many basic functions, including differentiation and development. In pathological conditions such as inflammation and tumorigenesis adhesive events also play a major role. Cellular adhesion is mediated by specific molecules expressed by both normal and neoplastic tissues. Capillary hemangioblastoma is a tumor of controversial origin, characterized by two major components, vacuolated stromal cells and a capillary network. In order to shed light on the differentiation of the stromal cells and the interactions between the two major components of hemangioblastoma we studied the expression of several adhesion molecules by immunocytochemistry. The endothelium-associated adhesion molecules (ICAM-1, ICAM-2, VCAM-1, PECAM-1 and ELAM-1) were expressed by endothelial cells within the tumors, but not by stromal cells. In contrast, the stromal cells showed strong neuronal cell adhesion molecule (NCAM/CD56) expression, further distinguishing them from endothelial cells. In addition, the stromal cells expressed CD44, which is of interest, as this membrane protein is linked to ezrin, a cytoskeleton-associated protein also expressed by stromal cells. We conclude that the stromal cells and endothelial cells of capillary hemangioblastoma exhibit quite divergent expression patterns of adhesion molecules. The NCAM expression in stromal cells suggests neuroectodermal or mesenchymal differentiation of this tumor. In addition, the NCAM expression could contribute to the sometimes problematic differential diagnosis between capillary hemangioblastoma and metastatic renal cell carcinoma of the central nervous system. Received: 17 December 1995 / Revised, accepted: 13 May 1996     

Inconsistent association of Epstein-Barr virus with CD56 (NCAM)-positive angiocentric lymphoma occuring in sites other than the upper and lower respiratory tract

We previously described nine cases of angiocentric lymphoma of a possible natural killer (NK)-cell lineage with a surface CD3− CD56+ phenotype occurring in sites other than the upper and lower respiratory tract. This study was performed to investigate the association of Epstein-Barr virus (EBV) with these lymphomas, using the polymerase chain reaction (PCR) for the presence of EBV-DNA, in situ hybridization (ISH) for EBV-encoded small RNAs (EBERs) and immunohistology for EBV-determined nuclear antigen-2 (EBNA-2) and latent membrane protein-1 (LMP-1) in paraffin sections. PCR and ISH produced almost identical results, and EBERs were identified in the nuclei of the lymphoma cells of three cases, two of which exhibited LMP-1 in the cytoplasm of tumour cells without EBNA-2 expression. Molecular genetic analysis revealed EBV to be incorporated into these three EBER-positive cases either clonally or biclonally. It was revealed by re-evaluation of their morphology with the established EBV status on each case that, in contrast to the rather variable and irregular cellular composition of the EBV- positive tumours, the EBV-negative tumours stood out because of their remarkably uniform 'blastoid' appearance, and could be grouped as blastic NK-cell lymphoma. The relationship of the EBV-positive cases with nasal NK-cell tumours has yet to be clarified.     

Interactions between growing thalamocortical afferent axons and the neocortical primordium in normal and reeler mutant mice

The interactions between growing thalamocortical afferent axons and the neocortical primordium were examined during neocortical development of the mouse cerebrum, by labeling the afferents with the carbocyanine fluorescent dye, DiI, which was introduced into the dorsal thalamus of the fixed brains of control and reeler mutant mice. In the neocortical primordium of the control mouse, the labeled afferents running tangentially in the intermediate zone formed a dense plexus in the subplate, the layer below the cortical plate, as early as the 16th gestational day (E16). Small numbers of the afferents invaded the lower cortical plate at E16 and increasing numbers of labeled growing axons extended into the cortical plate at E17. At the 4th postnatal day (P4), labeled afferents grew radially up to the upper cortical plate and terminal arborizations of the afferents were evident in the forming layer IV. In contrast, in the E16 cerebrum of the reeler mutant mouse, in which the cortical layers are inverted, the labeled afferents traversed the neocortical primordium directly towards the superplate, the superficial layer above the cortical plate and the equivalent of the subplate in the control mouse. Thick bundles of labeled axons reached the superplate and made contact with the superplate neurons. At P4 in the reeler neocortex, the afferent axons that had reached the superplate began to change their direction of growth and turned towards the deeper layer. Electron-microscopic observations at E16 revealed that immature synapses were formed on the somata of the subplate neurons in the control mouse, and similar immature synapses were also formed on the superplate neurons of the reeler mutant. At E16 in the control, NGF receptor immunoreactivity was expressed in the intermediate zone, subplate and lower cortical plate, and the mode of expression corresponded to the distribution of thalamocortical afferents. At the same stage of the reeler mutant, expression of NGF receptor immunoreactivity was confined to the afferent axons that had grown through the neocortical primordium towards the superplate. In the control at E17, highly polysialylated NCAM (NCAM-H), a homophilic cell adhesion molecule, was expressed in the subplate, marginal zone and afferent axons. In the reeler mutant at the same stage, this adhesion molecule was expressed in both the superplate and the bundles of the afferent axons. These findings suggest that the subplate and the superplate, which are composed of neurons generated at the earliest stage, attract growing thalamocortical afferent axons specifically by a chemotropic mechanism through the expression of NGF receptor. Furthermore, growth cones of the afferent axons may make contact with the subplate or superplate neurons specifically through the homophilic adhesive activity of NCAM-H expressed on them. On the basis of such mechanisms, the subplate or the superplate could play a role as a tentative target for the thalamocortical afferents prior to arrival at their final targets, i.e., the layer IV cortical neurons in the control and the equivalent neurons in the reeler mutant.     

NCAM-antibodies modulate induction of long-term potentiation in rat hippocampal CA1

The neural cell adhesion molecule (NCAM) probably plays a role in neural plasticity in the adult vertebrate brain. We here present evidence that NCAM may be involved in long-term potentiation (LTP) in the CA1-region of rat hippocampal slices. It is shown that local application of antibodies against NCAM inhibits subsequent LTP-induction. Thus NCAM may be directly involved in the initial phase of LTP-induction. These results have important implications for the possible involvement of NCAM in learning and memory.     

Effect of experimental hyperphenylalaninemia on the postnatal rat brain

The molecular mechanism of the disturbance to brain development caused by phenylketonuria remains mostly unknown. We have studied three molecular markers that reflect the development of neurons, glia and the extracellular matrix of the postnatal rat brain in an animal model of hyperphenylalaninemia, in order to elucidate the possible mechanism by which increased phenylalanine influences brain development. The contents of NCAM, GFAP and hyaluronate-binding activity were compared in cerebellum and telencephalon of normal rats and those subjected to high phenylalanine. No statistically significant changeswere found in telencephalon when experimental animals were compared to controls. In the hyperphenylalaninemic cerebellum, the developmental dynamic of NCAM content (represented by two peaks at about postnatal days 5 and 22 during normal development) is dramatically altered. The GFAP content in the cerebellum of treated rats exceeded those in controls significantly during late developmental stages (postnatal days 28–35). Hyaluronate-binding activity in the extracellular protein fraction from treated rat cerebellum was increased compared to normal rat at the early stages of development only (postnatal day 7). These results suggest that high serum phenylalanine may lead to permanent brain dysfunction through a disturbance of a wide range of developmental events.