Development Of Neuronal Connections Research Articles

Semaphorin 4C Plexin-B2 signaling in peripheral sensory neurons is pronociceptive in a model of inflammatory pain

Semaphorins and their transmembrane receptors, Plexins, are key regulators of axon guidance and development of neuronal connectivity. B-type Plexins respond to Class IV semaphorins and mediate a variety of developmental functions. Here we report that the expression of Plexin-B2 and its high-affinity ligand, Sema4C, persists in peripheral sensory neurons in adult life and is markedly increased in states of persistent pain in mice. Genetic deletion of Sema4C as well as adult-onset loss of Plexin-B2 leads to impairment of the development and duration of inflammatory hypersensitivity. Remarkably, unlike the neurodevelopmental functions of Plexin-B2 that solely rely on Ras signaling, we obtained genetic and pharmacological evidence for a requirement of RhoA-ROCK-dependent mechanisms as well as TRPA1 sensitization in pronociceptive functions of Sema4C-Plexin-B2 signaling in adult life. These results suggest important roles for Plexin-B2 signaling in sensory function that may be of therapeutic relevance in pathological pain.

The anatomy, organisation and development of contralateral callosal projections of the mouse somatosensory cortex.

Background:Alterations in the development of neuronal connectivity can result in dramatic outcomes for brain function. In the cerebral cortex, most sensorimotor and higher-order functions require coordination between precise regions of both hemispheres through the axons that form the corpus callosum. However, little is known about how callosal axons locate and innervate their contralateral targets.Methods:Here, we use a combination of in utero electroporation, retrograde tracing, sensory deprivation and high-resolution axonal quantification to investigate the development, organisation and activity dependence of callosal axons arising from the primary somatosensory cortex of mice.Results:We show that distinct contralateral projections arise from different neuronal populations and form homotopic and heterotopic circuits. Callosal axons innervate the contralateral hemisphere following a dorsomedial to ventrolateral and region-specific order. Furthermore, we identify two periods of region- and layer-specific developmental exuberance that correspond to initial callosal axon innervation and subsequent arborisation. Early sensory deprivation affects only the latter of these events.Conclusion:Taken together, these results reveal the main developmental events of contralateral callosal targeting and may aid future understanding of the formation and pathologies of brain connectivity.

Negative Allosteric Modulation of mGluR5 Partially Corrects Pathophysiology in a Mouse Model of Rett Syndrome.

Altered regulation of synaptic protein synthesis has been hypothesized to contribute to the pathophysiology that underlies multiple forms of intellectual disability and autism spectrum disorder. Here, we show in a mouse model of Rett syndrome (Mecp2 KO) that metabotropic glutamate receptor 5 (mGluR5)- and protein-synthesis-dependent synaptic plasticity are abnormal in the hippocampus. We found that a subset of ribosome-bound mRNAs was aberrantly upregulated in hippocampal CA1 neurons of Mecp2 KO mice, that these significantly overlapped with FMRP direct targets and/or SFARI human autism genes, and that chronic treatment of Mecp2 KO mice with an mGluR5-negative allosteric modulator tunes down upregulated ribosome-bound mRNAs and partially improves mutant mice phenotypes.

Molecular Mechanism of Use-Dependent Activation of Kv1.2 Channel Complexes and its Impact on Regulation of Neuronal Excitability

Voltage-gated potassium channels (Kv) play a critical role in the regulation of threshold and action potential morphology in virtually all excitable tissues. The Kv1.2 channel in particular plays a significant role in normal neuronal function. Genetic deletion of the Kv1.2 subunit is invariably lethal in mice shortly after birth, and in humans Kv1.2 channelopathies precipitate epileptic encephalopathy and ataxia. We have investigated a poorly recognized regulatory mechanism that profoundly affects Kv1.2 function, termed ‘use-dependent activation’, whereby repetitive stimulation leads to dramatic potentiation of channel current. The physiological significance of this observation is that potentiation of Kv1.2 function may act as a silencing mechanism in repetitively firing neurons. Our findings suggest that use-dependent activation is mediated by an extrinsic regulatory factor that binds Kv1.2 in a state-dependent manner. Point mutations of a putative binding region in the S2-S3 linker suggest important steric and charge requirements to allow interaction of the regulatory factor. While sensitivity is exclusive to Kv1.2 subunits, use-dependent activation can be harnessed in heteromeric channels containing at least one Kv1.2 subunit. We extend these observations by demonstrating that use-dependent activation persists in primary cultured neurons. Using tityustoxin for acute and long-term inhibition of Kv1.2-containing channels in neuronal cultures, we have investigated the role of Kv1.2 subunits on action potential firing and development of neuronal connectivity. Overall, we have dissected the sequence determinants of use-dependent activation of Kv1.2 channels, and made strides toward understanding the implications of this regulatory mechanism in neuronal function.

MicroRNAs and synaptic plasticity--a mutual relationship.

MicroRNAs (miRNAs) are rapidly emerging as central regulators of gene expression in the postnatal mammalian brain. Initial studies mostly focused on the function of specific miRNAs during the development of neuronal connectivity in culture, using classical gain- and loss-of-function approaches. More recently, first examples have documented important roles of miRNAs in plastic processes in intact neural circuits in the rodent brain related to higher cognitive abilities and neuropsychiatric disease. At the same time, evidence is accumulating that miRNA function itself is subjected to sophisticated control mechanisms engaged by the activity of neural circuits. In this review, we attempt to pay tribute to this mutual relationship between miRNAs and synaptic plasticity. In particular, in the first part, we summarize how neuronal activity influences each step in the lifetime of miRNAs, including the regulation of transcription, maturation, gene regulatory function and turnover in mammals. In the second part, we discuss recent examples of miRNA function in synaptic plasticity in rodent models and their implications for higher cognitive function and neurological disorders, with a special emphasis on epilepsy as a disorder of abnormal nerve cell activity.

A Precisely Timed Asynchronous Pattern of ON and OFF Retinal Ganglion Cell Activity during Propagation of Retinal Waves

Patterns of coordinated spontaneous activity have been proposed to guide circuit refinement in many parts of the developing nervous system. It is unclear, however, how such patterns, which are thought to indiscriminately synchronize nearby cells, could provide the cues necessary to segregate functionally distinct circuits within overlapping cell populations. Here, we report that glutamatergic retinal waves possess a substructure in the bursting of neighboring retinal ganglion cells with opposite light responses (ON or OFF). Within a wave, cells fire repetitive nonoverlapping bursts in a fixed order: ON before OFF. This pattern is absent from cholinergic waves, which precede glutamate-dependent activity, providing a developmental sequence of distinct activity-encoded cues. Asynchronous bursting of ON and OFF retinal ganglion cells depends on inhibition between these parallel pathways. Similar asynchronous activity patterns could arise throughout the nervous system, as inhibition matures and might help to separate connections of functionally distinct subnetworks.

Semaphorin-1a Controls Receptor Neuron-Specific Axonal Convergence in the Primary Olfactory Center of Drosophila

In the olfactory system of Drosophila, 50 functional classes of sensory receptor neurons (ORNs) project in a highly organized fashion into the CNS, where they sort out from one another and converge into distinct synaptic glomeruli. We identified the transmembrane molecule Semaphorin-1a (Sema-1a) as an essential component to ensure glomerulus-specific axon segregation. Removal of sema-1a in ORNs does not affect the pathfinding toward their target area but disrupts local axonal convergence into a single glomerulus, resulting in two distinct targeting phenotypes: axons either intermingle with adjacent ORN classes or segregate according to their odorant receptor identity into ectopic sites. Differential Sema-1a expression can be detected among neighboring glomeruli, and mosaic analyses show that sema-1a functions nonautonomously in ORN axon sorting. These findings provide insights into the mechanism by which afferent interactions lead to synaptic specificity in the olfactory system.

Mapping Labels in the Human Developing Visual System and the Evolution of Binocular Vision

Topographic representation of visual fields from the retina to the brain is a central feature of vision. The development of retinotopic maps has been studied extensively in model organisms and is thought to be controlled in part by molecular labels, including ephrin/Eph axon guidance molecules, displayed in complementary gradients across the retina and its targeting areas. The visual system in these organisms is primarily monocular, with each retina mapping topographically to its contralateral target. In contrast, mechanisms of retinal mapping in binocular species such as primates, characterized by the congruent, aligned mapping of both retinas onto the same brain target, remain completely unknown. Here, we show that the distribution of ephrin/Eph genes in the human developing visual system is fundamentally different from what is known in model organisms. In the human embryonic retina, EphA receptors are displayed along two gradients, sloping down from the center of the retina to its periphery. The EphB1 receptor, which controls the ipsilateral routing of retinal axons in the mouse, is expressed throughout the human temporal retina in coordination with the changes in EphA gene expression. In the dorsal lateral geniculate nucleus, ephrin-A/EphAs are displayed along complementary retinotopic gradients. Our data point to an evolutionary model in which the coordinated divergence of the distribution of the receptors controlling retinal guidance and retinal mapping enabled the emergence of a fully binocular system. They also indicate that ephrin/Eph signaling plays a potentially major role in the development of neuronal connectivity in humans.

Neurotrophic regulation of retinal ganglion cell synaptic connectivity: from axons and dendrites to synapses

This review highlights important events during the morphological development of retinal ganglion cells (RGCs), focusing on mechanisms that control axon and dendritic arborization as a means to understand synaptic connectivity with special emphasis on the role of neurotrophins during structural and functional development of RGCs. Neurotrophins and their receptors participate in the development of visual connectivity at multiple levels. In the visual system, neurotrophins have been shown to exert various developmental influences, from guiding the morphological differentiation of neurons to controlling the functional plasticity of visual circuits. This review article examines the role of neurotrophins, and in particular of BDNF, during the morphological development of RGCs, and discusses potential interactions between activity and neurotrophins during development of neuronal connectivity.

Insulin Receptors and Visual Development

In Drosophila , the insulin receptor (DInR) is expressed throughout the animal. Song et al. (see the Perspective by Dickson) now analyze how the DInR functions during visual system development. As axons make their way from the retina to their final targets in the brain, a well-organized mapping results in thorough distribution of retinal connections across the optic lobe. When DInR or its adaptor protein Dock are disrupted by mutation, the retinotectal axonal connections are also disrupted. Thus, in Drosophila , one of the myriad functions of the insulin receptor is to mediate, with the help of Dock, the normal development of neuronal connections from eye to brain. J. Song, L. Wu, Z. Chen, R. A. Kohanski, L. Pick, Axons guided by insulin receptor in Drosophila visual system. Science 300 , 502-505 (2003). [Abstract] [Full Text] B. J. Dickson, Wiring the brain with insulin. Science 300 , 440-441 (2003). [Summary] [Full Text]

Retinal axon growth cones respond to EphB extracellular domains as inhibitory axon guidance cues.

Axon pathfinding relies on cellular signaling mediated by growth cone receptor proteins responding to ligands, or guidance cues, in the environment. Eph proteins are a family of receptor tyrosine kinases that govern axon pathway development, including retinal axon projections to CNS targets. Recent examination of EphB mutant mice, however, has shown that axon pathfinding within the retina to the optic disc is dependent on EphB receptors, but independent of their kinase activity. Here we show a function for EphB1, B2 and B3 receptor extracellular domains (ECDs) in inhibiting mouse retinal axons when presented either as substratum-bound proteins or as soluble proteins directly applied to growth cones via micropipettes. In substratum choice assays, retinal axons tended to avoid EphB-ECDs, while time-lapse microscopy showed that exposure to soluble EphB-ECD led to growth cone collapse or other inhibitory responses. These results demonstrate that, in addition to the conventional role of Eph proteins signaling as receptors, EphB receptor ECDs can also function in the opposite role as guidance cues to alter axon behavior. Furthermore, the data support a model in which dorsal retinal ganglion cell axons heading to the optic disc encounter a gradient of inhibitory EphB proteins which helps maintain tight axon fasciculation and prevents aberrant axon growth into ventral retina. In conclusion, development of neuronal connectivity may involve the combined activity of Eph proteins serving as guidance receptors and as axon guidance cues.

All-or-none potentiation at CA3-CA1 synapses.

The molecular mechanisms underlying long-term potentiation in the hippocampus have received much attention because of the likely functional importance of synaptic plasticity for information storage and the development of neuronal connectivity. Surprisingly, it remains unclear whether activity modifies the strength of individual synapses in a digital (all-or-none) or analog (graded) manner. Here we characterize step-like all-or-none transitions from baseline synaptic transmission to potentiated states following protocols for inducing potentiation at putative single CA3-CA1 synaptic connections. Individual synapses appear to have all-or-none potentiation indicative of highly cooperative processes but different thresholds for undergoing potentiation. These results raise the possibility that some forms of synaptic memory may be stored in a digital manner in the brain.

Labeling Neural Cells Using Adenoviral Gene Transfer of Membrane-Targeted GFP

We describe an experimental system to visualize the soma and processes of mammalian neurons and glia in living and fixed preparations by using a recombinant adenovirus vector to transfer the jellyfish green fluorescent protein (GFP) into postmitotic neural cells both in vitro and in vivo. We have introduced several modifications of GFP that enhance its fluorescence intensity in mammalian axons and dendrites. This method should be useful for studying the dynamic processes of cell migration and the development of neuronal connections, as well as for analyzing the function of exogenous genes introduced into cells using the adenovirus vector.

Development of neuronal connections. Symposium of the Cajal Club. April 6, 1986, Reno, Nevada. Proceedings.

Development of neuronal connections. Symposium of the Cajal Club. April 6, 1986, Reno, Nevada. Proceedings.

Development of neuronal connections between a transplanted lateral geniculate nucleus and the host visual cortex in rats

Development of neuronal connections between a transplanted lateral geniculate nucleus and the host visual cortex in rats

A role for action-potential activity in the development of neuronal connections in the kitten retinogeniculate pathway

The role of action potentials in the development of proper synaptic connections in the mammalian CNS was studied in the kitten retinogeniculate pathway. Our basic finding is that there is improper segregation of retinal inputs onto LGN cells after prolonged retinal action-potential blockade. Retinal ganglion cell firing was silenced from birth by repeated monocular injections of TTX. The resulting ganglion cell connections in the LGN were studied electrophysiologically after the action-potential blockade was ended. Most cells in the deprived LGN layers received excitatory input from both ON-center and OFF-center type ganglion cells, whereas LGN cells normally receive inputs only from ON-center or OFF-center ganglion cells, but not from both types. Improper segregation of ON and OFF inputs has never been reported after other types of visual deprivation that do not block ganglion cell activity. Control experiments showed that receptive fields in the nondeprived LGN layers were normal, that ganglion cell responses remained normal, and that there was no obvious ganglion cell loss. We also showed that individual LGN cells with ON and OFF excitatory inputs were not present in normal neonatal kittens. Two other types of improper input segregation in response to action-potential blockade were also found in the deprived LGN layers. (1) A greater than normal number of LGN cells received both X- and Y-type ganglion cell input. (2) Almost half of the cells at LGN layer borders were excited binocularly. Recovery of LGN normality was rapid and complete after blockade that lasted for only 3 weeks from birth, but little recovery was seen after about 11 weeks of blockade. The susceptibility to action-potential blockade decreased during the first 3 postnatal weeks. These findings may result from axon-terminal sprouting or from the failure of axon terminals to retract. The results are consistent with the idea that normally synchronous activity of neighboring ganglion cells of like center-type may be used in the refinement of retinogeniculate synaptic connections.

Postnatal changes in the number of astrocytes, oligodendrocytes, and microglia in the visual cortex (area 17) of the macaque monkey: a stereological analysis in normal and monocularly deprived animals.

The numerical densities (Nv) of astrocytes, oligodendrocytes, and microglia were measured in individual laminae of the striate cortex of macaque monkeys ranging in age from newborn to adult. Using measurements of cortical thickness and surface area, the total number of cells in the striate cortex of one hemisphere was derived for each glial cell type. Normal monkeys were compared at 3 months and 6 months of age to animals reared from birth with a monocular eyelid suture. No significant differences were observed between normal and monocularly deprived monkeys. The combined data from these groups, however, demonstrated several significant developmental changes. The Nv of astrocytes decreased from birth to 6 months of age and subsequently increased in the adult. The greatest changes were seen in the more superficial laminae. These changes, however, were only a response to a substantial overshoot in cortical volume at six months: the total number of astrocytes in the striate cortex did not change. There was a tenfold increase in both the Nv and the total number of oligodendrocytes from birth to maturity with a corresponding increase in the density of myelinated axons. The greatest changes were observed in the deeper laminae. The total number of microglia remained relatively constant from birth to 6 months of age. There was a 55% reduction in the number of microglia in the adult, although statistical analysis indicated that this decrease was only of borderline significance. The possible relationships between these postnatal changes in glial cell numbers and the development of neuronal connectivity are discussed.

Development of neuronal connections in chick embryonic retino-tectal system: An overview

Synaptogenesis has been examined in the retino-tectal system of the chick embryo both biochemically and morphologically. We have evaluated the amount and the rate transport of glycoproteins synthetized in retinal ganglion cells and axonally transported to retinal nerve endings. The distribution of glycoproteins inside the tectum shows that they are part of the synaptosomal membrane either because they are incorporated in it "en route" or because they flow through an axolemmal flow. The role they might play in the building up of the synaptic membrane is discussed in relation to the maturation of synaptic contacts, observed either with E-PTA staining method or with routine fixation methods. The distribution of a carbohydrate binding protein was followed with histochemical techniques. This protein is synthesized by the matching tectal neurons, and its synthesis is developmentally regulated. It distribution is affected by innervation. All of these findings are discussed in relation to the hypothesis that three different steps are involved in embryonic synaptogenesis. These three steps are: 1) interneuronal recognition; 2) formation of initial contact, and 3) formation of synapses. The possibility that different kinds of cell surface macromolecules might play a different role is also discussed.

Structure and development of neuronal connections in isogenic organisms: transient gap junctions between growing optic axons and lamina neuroblasts.

We previously showed that the growth of each bundle of eight optic fibers from one ommatidium into the optic lamina of Daphnia occurs in such a way that one of the eight fibers precedes the others into the lamina. The growth cone of this lead fiber makes surface contact with undifferentiated neuroblasts near the midplane. This is followed by a glial-like wrapping of each neuroblast around the fiber. In this report, gap junctions are shown to form for a short period of time between the growing lead fiber and the neuroblast that is wrapping around it. It is proposed that these junctions may represent a morphological correlate of informational exchange between axon and neuroblast. This signaling would then reflect the fact that the sequence of axon proliferation by the lamina neuroblasts within an optic cartrdige, ultimately composed of five lamina neurons and eight optic fibers, parallels the order in which the neuroblasts undergo the wrapping reaction with the lead fiber.

Structure and development of neuronal connections in isogenic organisms: cellular interactions in the development of the optic lamina of Daphnia.

Some details of the growth and initial cellular interactions of optic nerve axons were examined in a parthenogenetic clone of Daphnia magna. Results are summarized as follows: (i) the final structure of the optic lamina is dependent upon interactions between growing optic nerve fibers and optic lamina neuroblasts closest to the midplane of the animal, which trigger the morphological differentiation of the neuroblasts; the specificity of connections is achieved by well-defined sequences of cell migration in the ganglion; (ii) only one of the eight optic nerve axons growing back from each ommatidium in the eye possesses a structure similar to the growth cones seen on termini of nerve fibers growing in vitro; and (iii) undifferentiated neuroblasts in the ganglion react to surface contact by this "lead axon" by enveloping the axon in a glial-like relationship.