Cadherins in neuronal morphogenesis and function

Classic cadherins represent a family of calcium-dependent homophilic cell–cell adhesion molecules. They confer strong adhesiveness to animal cells when they are anchored to the actin cytoskeleton via their cytoplasmic binding partners, catenins. The cadherin/catenin adhesion system plays key roles in the morphogenesis and function of the vertebrate and invertebrate nervous systems. In early vertebrate development, cadherins are involved in multiple events of brain morphogenesis including the formation and maintenance of the neuroepithelium, neurite extension and migration of neuronal cells. In the invertebrate nervous system, classic cadherin-mediated cell–cell interaction plays important roles in wiring among neurons. For synaptogenesis, the cadherin/catenin system not only stabilizes cell–cell contacts at excitatory synapses but also assembles synaptic molecules at synaptic sites. Furthermore, this system is involved in synaptic plasticity. Recent studies on the role of individual cadherin subtypes at synapses indicate that individual cadherin subtypes play their own unique role to regulate synaptic activities.     

Dendritic Spine Loss and Synaptic Alterations in Alzheimer’s Disease

Dendritic spines are tiny protrusions along dendrites, which constitute major postsynaptic sites for excitatory synaptic transmission. These spines are highly motile and can undergo remodeling even in the adult nervous system. Spine remodeling and the formation of new synapses are activity-dependent processes that provide a basis for memory formation. A loss or alteration of these structures has been described in patients with neurodegenerative disorders such as Alzheimer's disease (AD), and in mouse models for these disorders. Such alteration is thought to be responsible for cognitive deficits long before or even in the absence of neuronal loss, but the underlying mechanisms are poorly understood. This review will describe recent findings and discoveries on the loss or alteration of dendritic spines induced by the amyloid beta (Abeta) peptide in the context of AD.     

Wnt Signaling in the Vertebrate Central Nervous System: From Axon Guidance to Synaptic Function

Regulation of cell signaling by Wnt proteins is critical for the formation of neuronal circuits. Wnts modulate axon pathfinding, dendritic development, and synaptic assembly. Through different receptors, Wnts activate diverse signaling pathways that lead to local changes on the cytoskeleton or global cellular changes involving nuclear function. Recently, a link between neuronal activity, essential for the formation and refinement of neuronal connections, and Wnt signaling has been uncovered. Indeed, neuronal activity regulates the release of Wnt and the localization of their receptors. Wnts mediate synaptic structural changes induced by neuronal activity or experience. New emerging evidence suggests that dysfunction in Wnt signaling contributes to neurological disorders. In this article, the attention is focused on the function of Wnt signaling in the formation of neuronal circuits in the vertebrate central nervous system.The formation of neuronal connections requires the navigation of axons to their appropriate synaptic targets, the formation of terminal branches, and the assembly of functional synapses. These processes greatly depend on the proper dialogue between axons and their environment as they navigate to their target, and between axons and their postsynaptic dendrites during synapse assembly. A combination of secreted molecules and transmembrane proteins modulates these processes. Studies over the last 10 years have revealed an essential role for Wnt signaling in axon pathfinding, dendritic development, and synapse assembly in both central and peripheral nervous systems. Wnts also modulate basal synaptic transmission and the structural and functional plasticity of synapses in the central nervous system. Studies of Wnts in the nervous system have significantly contributed to our current understanding of the molecular mechanisms that control neuronal circuit assembly. These studies have also shed light into fundamental aspects of cell signaling such as novel mechanisms of protein secretion (Korkut et al. 2009) and receptor dynamics (Sahores et al. 2010). Here I review the mechanisms by which Wnts modulate axon guidance and synapse formation in the vertebrate central nervous system. I also discuss the increasing evidence in support for a role of Wnts in basal synaptic transmission, synaptic plasticity, and neurological disorders.     

Eph/ephrin对树突棘的调控与中枢神经系统疾病

树突棘是神经元树突上的功能性突起结构,通常作为突触后成份与投射来的轴突共同构成完整的突触连接。树突棘的形态与结构具有明显的可塑性,其变化通常会引起突触功能的改变。Eph受体酪氨酸激酶家族分子与其配体ephrin都是重要的神经导向因子,同时对树突棘结构也有直接的调控作用。Eph受体的活化可以促进树突棘的发生并影响树突棘的形态及内部结构;而Eph受体的异常也往往会损害正常的突触功能,甚至导致许多与树突棘结构异常相关的神经系统病变的发生。     

Developmental regulation of axon branching in the vertebrate nervous system

During nervous system development, axons generate branches to connect with multiple synaptic targets. As with axon growth and guidance, axon branching is tightly controlled in order to establish functional neural circuits, yet the mechanisms that regulate this important process are less well understood. Here, we review recent advances in the study of several common branching processes in the vertebrate nervous system. By focusing on each step in these processes we illustrate how different types of branching are regulated by extracellular cues and neural activity, and highlight some common principles that underlie the establishment of complex neural circuits in vertebrate development.     

外周输入依赖的嗅球颗粒细胞的突触结构可塑性

活动依赖的突触结构可塑性是学习和记忆的基础.哺乳动物,尤其是啮齿类动物,具有高度发达的嗅觉系统和惊人的气味学习和记忆能力.本研究以CNGA2敲除而导致外周输入缺失的小鼠为模型,研究嗅球内活动依赖的突触结构可塑性.利用特异性的突触前和突触后标记物,发现外周输入缺失减少了突触标记蛋白突触素(synaptophysin)和抑制性突触标记蛋白桥蛋白(gephyrin)在嗅球外网状层和颗粒细胞层中的表达;兴奋性突触标记蛋白囊泡谷氨酸转运蛋白1(VGluT1)的表达水平只在外网状层中有显著下降,而在颗粒细胞层中没有明显变化.进一步通过活体质粒电转标记嗅球颗粒细胞后发现,CNGA2敲除小鼠颗粒细胞上位于外网状层中的远端树突棘密度显著减小,而位于颗粒细胞层中的近端树突棘密度没有明显变化.这些结果表明颗粒细胞上的树-树突触具有对外周活动依赖的结构可塑性,而轴-树突触则无.     

Do thin spines learn to be mushroom spines that remember?

Dendritic spines are the primary site of excitatory input on most principal neurons. Long-lasting changes in synaptic activity are accompanied by alterations in spine shape, size and number. The responsiveness of thin spines to increases and decreases in synaptic activity has led to the suggestion that they are 'learning spines', whereas the stability of mushroom spines suggests that they are 'memory spines'. Synaptic enhancement leads to an enlargement of thin spines into mushroom spines and the mobilization of subcellular resources to potentiated synapses. Thin spines also concentrate biochemical signals such as Ca(2+), providing the synaptic specificity required for learning. Determining the mechanisms that regulate spine morphology is essential for understanding the cellular changes that underlie learning and memory.     

BDNF mobilizes synaptic vesicles and enhances synapse formation by disrupting cadherin-beta-catenin interactions

Neurons of the vertebrate central nervous system have the capacity to modify synapse number, morphology, and efficacy in response to activity. Some of these functions can be attributed to activity-induced synthesis and secretion of the neurotrophin brain-derived neurotrophic factor (BDNF); however, the molecular mechanisms by which BDNF mediates these events are still not well understood. Using time-lapse confocal analysis, we show that BDNF mobilizes synaptic vesicles at existing synapses, resulting in small clusters of synaptic vesicles "splitting" away from synaptic sites. We demonstrate that BDNF's ability to mobilize synaptic vesicle clusters depends on the dissociation of cadherin-beta-catenin adhesion complexes that occurs after tyrosine phosphorylation of beta-catenin. Artificially maintaining cadherin-beta-catenin complexes in the presence of BDNF abolishes the BDNF-mediated enhancement of synaptic vesicle mobility, as well as the longer-term BDNF-mediated increase in synapse number. Together, this data demonstrates that the disruption of cadherin-beta-catenin complexes is an important molecular event through which BDNF increases synapse density in cultured hippocampal neurons.     

Demonstration of insulin-related substances in the central nervous systems of pulmonates and Aplysia californica

Summary the occurrence of insulin-related substances in the central nervous system of pulmonates and Aplysia californica was investigated by means of immunocytochemistry and in situ hybridization. Previous experiments have shown that, in Lymnaea stagnalis, the growth hormone-producing neurons in the cerebral ganglia (the so-called light green cells) express at least 5 genes that are related to the vertebrate insulin genes, i.e., they encode prohormones that are composed of a B- and A-chain and a connecting C peptide. These insulin related molecules also have the amino acids essential for their tertiary structure (viz. cysteines) at identical positions to those of the vertebrate insulins. In the investigated basommatophoran and stylommatophoran snails and slugs, neurons reacted with an antiserum raised against the C peptide of one of the molluscan insulin-related peptides. These neurons can be considered to be, based on morphological and endocrinological criteria, homologous to the light green cells of L. stagnalis. In A. californica, all central ganglia contain immunoreactive neurons. The highest number (about 50) was observed in the abdominal ganglion. The present results indicate that insulin-related substances are generally occurring neuropeptides in the central nervous system of molluscs.     

Ultrastructure of the synapses of the initial axonal segment of the pyramidal neuron

Ultrastructure of the proximal part of the axon in the neurons, identified according to a number of morphological signs as pyramidal, has been studied in the layer III of the cat cerebral hemisphere sensomotor cortex. In sections, tangential to the cortical surface, in the initial axonal segment, a submembranous osmophilic layer and fasciculi of microtubules are revealed. On the initial segment spines are found, they contain cysterns resembling by their structure the spine system of the dendritic spines. Axonal terminals revealed along the axonal distribution are in contact both with the axonal trunk and with the spines. Regarding the initial segment, they are presynaptic, contain oval synaptic vesicles and form symmetric axo-axonal synapses only. In transversal sections axonal terminals are detected, arranging on the surface of the initial segment mostly as single ones, in longitudinal sections they are seen as clusters. Analysing the author's data and those from the literature, a conclusion is made that in intact animals the synaptic contacts at the initial segment of the axon are the only form of axo-axonal synapses in the neocortex.     

Neurexin-1 is required for synapse formation and larvae associative learning in Drosophila

Neurexins are highly polymorphic cell-surface adhesive molecules in neurons. In cultured mammalian cell system, they were found to be involved in synaptogenesis. Here, we report for the first time that Drosophila neurexin is required for synapse formation and associative learning in larvae. Drosophila genome encodes a single functional neurexin (CG7050; Neurexin-1 or Nrx-1), which is a homolog of vertebrate alpha-neurexin. Neurexin-1 is expressed in central nervous system and highly enriched in synaptic regions of the ventral ganglion and brain. Neurexin-1 null mutants are viable and fertile, but have shortened lifespan. The synapse number is decreased in central nervous system in Neurexin-1 null mutants. In addition, Neurexin-1 null mutants exhibit associative learning defect in larvae.     

Looking forward to EphB signaling in synapses

Eph receptors and their ligands ephrins comprise a complex signaling system with diverse functions that span a wide range of tissues and developmental stages. The variety of Eph receptor functions stems from their ability to mediate bidirectional signaling through trans-cellular Eph/ephrin interactions. Initially thought to act by directing repulsion between cells, Ephs have also been demonstrated to induce and maintain cell adhesive responses at excitatory synapses in the central nervous system. EphB receptors are essential to the development and maintenance of dendritic spines, which accommodate the postsynaptic sites of most glutamatergic excitatory synapses in the brain. Functions of EphB receptors are not limited to control of the actin cytoskeleton in dendritic spines, as EphB receptors are also involved in the formation of functional synaptic specializations through the regulation of glutamate receptor trafficking and functions. In addition, EphB receptors have recently been linked to the pathophysiology of Alzheimer's disease and neuropathic pain, thus becoming promising targets for therapeutic interventions. In this review, we discuss recent findings on EphB receptor functions in synapses, as well as the mechanisms of bidirectional trans-synaptic ephrin-B/EphB receptor signaling that shape dendritic spines and influence post-synaptic differentiation.     

LTP promotes a selective long-term stabilization and clustering of dendritic spines

Dendritic spines are the main postsynaptic site of excitatory contacts between neurons in the central nervous system. On cortical neurons, spines undergo a continuous turnover regulated by development and sensory activity. However, the functional implications of this synaptic remodeling for network properties remain currently unknown. Using repetitive confocal imaging on hippocampal organotypic cultures, we find that learning-related patterns of activity that induce long-term potentiation act as a selection mechanism for the stabilization and localization of spines. Through a lasting N-methyl-D-aspartate receptor and protein synthesis–dependent increase in protrusion growth and turnover, induction of plasticity promotes a pruning and replacement of nonactivated spines by new ones together with a selective stabilization of activated synapses. Furthermore, most newly formed spines preferentially grow in close proximity to activated synapses and become functional within 24 h, leading to a clustering of functional synapses. Our results indicate that synaptic remodeling associated with induction of long-term potentiation favors the selection of inputs showing spatiotemporal interactions on a given neuron.     

Changes in synaptic efficacy and seizure susceptibility in rat brain slices following extremely low‐frequency electromagnetic field exposure

The effects of electromagnetic fields (EMFs) on living organisms are recently a focus of scientific interest, as they may influence everyday life in several ways. Although the neural effects of EMFs have been subject to a considerable number of investigations, the results are difficult to compare since dissimilar exposure protocols have been applied on different preparations or animals. In the present series of experiments, whole rats or excised rat brain slices were exposed to a reference level‐intensity (250–500 µT, 50 Hz) EMF in order to examine the effects on the synaptic efficacy in the central nervous system. Electrophysiological investigation was carried out ex vivo, on neocortical and hippocampal slices; basic synaptic functions, short‐ and long‐term plasticity and seizure susceptibility were tested. The most pronounced effect was a decrease in basic synaptic activity in slices treated directly ex vivo observed as a diminution in amplitude of evoked potentials. On the other hand, following whole‐body exposure an enhanced short‐ and long‐term synaptic facilitation in hippocampal slices and increased seizure susceptibility in neocortical slices was also observed. However, these effects seem to be transient. We can conclude that ELF‐EMF exposure exerts significant effects on synaptic activity, but the overall changes may strongly depend on the synaptic structure and neuronal network of the affected region together with the specific spatial parameters and constancy of EMF. Bioelectromagnetics 30:631–640, 2009. © 2009 Wiley‐Liss, Inc.     

Wnt Signaling through the Ror Receptor in the Nervous System

The receptor tyrosine kinase-like orphan receptor (Ror) proteins are conserved tyrosine kinase receptors that play roles in a variety of cellular processes that pattern tissues and organs during vertebrate and invertebrate development. Ror signaling is required for skeleton and neuronal development and modulates cell migration, cell polarity, and convergent extension. Ror has also been implicated in two human skeletal disorders, brachydactyly type B and Robinow syndrome. Rors are widely expressed during metazoan development including domains in the nervous system. Here, we review recent progress in understanding the roles of the Ror receptors in neuronal migration, axonal pruning, axon guidance, and synaptic plasticity. The processes by which Ror signaling execute these diverse roles are still largely unknown, but they likely converge on cytoskeletal remodeling. In multiple species, Rors have been shown to act as Wnt receptors signaling via novel non-canonical Wnt pathways mediated in some tissues by the adapter protein disheveled and the non-receptor tyrosine kinase Src. Rors can either activate or repress Wnt target expression depending on the cellular context and can also modulate signal transduction by sequestering Wnt ligands away from their signaling receptors. Future challenges include the identification of signaling components of the Ror pathways and bettering our understanding of the roles of these pleiotropic receptors in patterning the nervous system.     

Mapping of experimental axon degeneration by electron microscopy of golgi preparations

Summary For the mapping of the terminal area of transected axons within the central nervous system, electron microscopy has recently been adopted. A greater accuracy is thereby obtained than with silver impregnation and light microscopy, since it becomes possible to determine the kinds of structure (soma, dendrites, spines) with which the degenerating boutons establish synaptic contact. In the present study this technique was extended by Golgi impregnation of such material with the aim of making possible classification of the postsynaptic neuron. A few days after transection of a pathway (commissural fibres to the hippocampus being used as a model in this study) the brain was fixed by perfusion with phosphate buffered formalin with sucrose. This was followed by immersion in an osmium tetroxide-potassium dichromate mixture (Dalton's fixative without sodium chloride) later replaced by a solution of silver nitrate. Satisfactory Golgi impregnation of nerve cells and processes was obtained. By careful trimming, and reembedding of selected areas, blocks for ultramicrotomy could be obtained which contained only one type of impregnated cell, e.g., hippocampal pyramidal cells.The relation of basal dendrites of such cells to degenerating boutons of commissural fibres was studied. Numerous examples of contact between degenerating boutons and spines belonging to the basal dendrites were seen. Although the Golgi precipitate obscured the postsynaptic substance in the spines, a number of features at the sites of contact were considered strong indication that many of the contacts were synapses and not merely the result of random juxtaposition. This combined procedure is supposed to be applicable to other problems and to other parts of the nervous system as well.This study was supported in part by Grant NB 02215 from the National Institute of Neurological Diseases and Blindness, U.S. Public Health Service. This ais is gratefully acknowledged. The author is indebted to Mrs. J. L. Vaaland and Mr. B. V. Johansen for valuable technical assistance.     

Molluscan neurons in culture: shedding light on synapse formation and plasticity

From genes to behaviour, the simple model system approach has played many pivotal roles in deciphering nervous system function in both invertebrates and vertebrates. However, with the advent of sophisticated imaging and recording techniques enabling the direct investigation of single vertebrate neurons, the utility of simple invertebrate organisms as model systems has been put to question. To address this subject meaningfully and comprehensively, we first review the contributions made by invertebrates in the field of neuroscience over the years, paving the way for similar breakthroughs in higher animals. In particular, we focus on molluscan (Lymnaea, Aplysia, and Helisoma) and leech (Hirudo) models and the pivotal roles they have played in elucidating mechanisms of synapse formation and plasticity. While the ultimate goal in neuroscience is to understand the workings of the human brain in both its normal and diseased states, the sheer complexity of most vertebrate models still makes it difficult to define the underlying principles of nervous system function. Investigators have thus turned to invertebrate models, which are unique with respect to their simple nervous systems that are endowed with a finite number of large, individually identifiable neurons of known function. We start off by discussing in vivo and semi-intact preparations, regarding their amenability to simple circuit analysis. Despite the 'simplicity' of invertebrate nervous systems however, it is still difficult to study individual synaptic connections in detail. We therefore emphasize in the next section, the utility of studying identified invertebrate neurons in vitro, to directly examine the development, specificity, and plasticity of synaptic connections in a well-defined environment, at a resolution that it is still unapproachable in the intact brain. We conclude with a discussion of the future of invertebrates in neuroscience in elucidating mechanisms of neurological disease and developing neuron-silicon interfaces.     

Morphological plasticity of rodent astroglia

In the past two decades, there has been an explosion of research on the role of neuroglial interactions in the control of brain homeostasis in both physiological and pathological conditions. Astrocytes, a subtype of glia in the central nervous system, are dynamic signaling elements that regulate neurogenesis and development of brain circuits, displaying intimate dynamic relationships with neurons, especially at synaptic sites where they functionally integrate the tripartite synapse. When astrocytes are isolated from the brain and maintained in culture, they exhibit a polygonal shape unlike their precursors in vivo. However, cultured astrocytes can be induced to undergo morphological plasticity leading to process formation, either by interaction with neurons or by the influence of pharmacological agents. This review highlights studies on the molecular mechanisms underlying morphological plasticity in astrocyte cultures and intact brain tissue, both in situ and in vivo.     

Dynamics of dendritic spines and their afferent terminals: spines are more motile than presynaptic boutons

Previous work has established that dendritic spines, sites of excitatory input in CNS neurons, can be highly dynamic, in later development as well as in mature brain. Although spine motility has been proposed to facilitate the formation of new synaptic contacts, we have reported that spines continue to be dynamic even if they bear synaptic contacts. An outstanding question related to this finding is whether the presynaptic terminals that contact dendritic spines are as dynamic as their postsynaptic targets. Using multiphoton time-lapse microscopy of GFP-labeled Purkinje cells and DiI-labeled granule cell parallel fiber afferents in cerebellar slices, we monitored the dynamic behavior of both presynaptic terminals and postsynaptic dendritic spines in the same preparation. We report that while spines are dynamic, the presynaptic terminals they contact are quite stable. We confirmed the relatively low levels of presynaptic terminal motility by imaging parallel fibers in vivo. Finally, spine motility can occur when a functional presynaptic terminal is apposed to it. These analyses further call into question the function of spine motility, and to what extent the synapse breaks or maintains its contact during the movement of the spine.     

The presynaptic compartment: signals and targets

Synaptic terminals are key elements in the functional and structural organization of the nervous system. Release of neurotransmitters, i.e. the activity specifically localized at the terminals, not only sustains the transfer of information among adjacent cells, but also contributes significantly to directing the non-random distribution of macromolecules in the plasmalemma of postsynaptic neurons, with major consequences in their general architecture (assembly of postsynaptic densities, dendritic spines, etc.). In order for these specific functions to be carried out, synaptic terminals need to be specialized in a variety of aspects with respect to the rest of the neuron. This minireview is specifically focused on two such aspects, the generation of transduction signals and their mechanism of action on intraterminal targets. In either aspects nerve terminals are by no means fully homogeneous, yet they certainly share a number of common features. These include the predominant role of Ca²⁺, collaborating however with other second messengers (cAMP, IP₃, diacylglycerol) in the control of processes such as transmitter release and its modulation.