Disruption of retinogeniculate afferent segregation by antagonists to NMDA receptors.

Afferent activity has an important role in the formation of connections in the developing mammalian visual system. But the extent to which the activity of target neurons shapes patterns of afferent termination and synaptic contact is not known. In the ferret's visual pathway, retinal ganglion cell axons from each eye segregate early in development into eye-specific laminae in the lateral geniculate nucleus (LGN). The dorsal laminae (termed laminae A and A1) then segregate further into inner and outer sublaminae that retain input from on-centre and off-centre retinal axons, respectively. Thus, individual retinogeniculate axons form terminal arbors within laminae A and A1 that are restricted to one inner or outer sublamina. We report here that blockade of N-methyl-D-aspartate (NMDA) receptors on LGN cells with specific antagonists during the period of sublamina formation prevents retinal afferents from segregating into 'On' and 'Off' sublaminae. Retinogeniculate axons have arbors that are not restricted appropriately, or are restricted in size but inappropriately positioned within the eye-specific laminae. NMDA receptor antagonists may specifically disrupt a mechanism by which LGN neurons detect correlated afferent and target activity, and have been shown to reduce retinogeniculate transmission more generally, causing LGN cells to have markedly reduced levels of activity. These results therefore indicate that the activity of postsynaptic cells can significantly influence the patterning of inputs and the structure of presynaptic afferents during development.     

Pathway selection by growth cones of identified motoneurones in live zebra fish embryos

How is the adult pattern of connections between motoneurones and the muscles that they innervate established during vertebrate development? Populations of motoneurones are thought to follow one of two patterns of development: (1) motor axons initially follow stereotyped pathways and project to appropriate regions of the developing muscle or (2) motor axons initially project to some regions that are incorrect, the inappropriate projections being eliminated subsequently. Here we observed individually identified motoneurones in live zebra fish embryos as they formed growth cones and as their growth cones navigated towards their targets. We report that from axogenesis, each motor axon followed a stereotyped pathway and projected only to the specific region of the muscle appropriate for its adult function. In addition, the peripheral arbor established by each motoneurone was restricted to a stereotyped region of its own segment and did not overlap with the peripheral arbor of the other motoneurones in that segment. We conclude that the highly stereotyped pattern of innervation seen in the adult is due to initial selection of the appropriate pathway, rather than elimination of incorrect projections.     

Modification of retinal ganglion cell axon morphology by prenatal infusion of tetrodotoxin

The cellular mechanisms by which the axons of individual neurons achieve their precise terminal branching patterns are poorly understood. In the visual system of adult cats, retinal ganglion cell axons from each eye form narrow cylindrical terminal arborizations restricted to alternate non-overlapping layers within the lateral geniculate nucleus (LGN). During prenatal development, axon arborizations from the two eyes are initially simple in shape and are intermixed with each other; they then gradually segregate to form complex adult-like arborizations in separate eye-specific layers by birth. Here we report that ganglion cell axons exposed to tetrodotoxin (TTX) to block neuronal activity during fetal life fail to form the normal pattern of terminal arborization. Individual TTX-treated axon arborizations are not stunted in their growth, but instead produce abnormally widespread terminal arborizations which extend across the equivalent of approximately two eye-specific layers. These observations suggest that during fetal development of the central nervous system, the formation of morphologically appropriate and correctly located axon terminal arborizations within targets is brought about by an activity-dependent process.     

Identified neurones isolated from leech CNS make selective connections in culture.

Neurones cultured in vitro offer distinct advantages for studying how processes grow towards their targets and form synaptic connections. In contrast to the complex events occurring during the development of the nervous system, synapse formation in culture can be analysed in a few neurones at a time and under controlled conditions. We have now dissected out and cultured single identified neurones from the central nervous system (CNS) of the adult leech. Various types of sensory cells, motor cells, and interneurones can be identified in leech ganglia--each with a stereotyped set of properties, including: (1) the electrical characteristics of its membrane, (2) the arborisation of its branches and the morphology of its terminals and (3) the pattern of connections it makes with other identified neurones, skin or muscle. Thus, cultured cells can be compared in detail with their counterparts in situ. We have found that isolated cells survive for several weeks, maintain their membrane properties, sprout and form selective connections.     

Synapse elimination in neonatal rat muscle is sensitive to pattern of muscle use

The synaptic connections among the cells of the vertebrate nervous system undergo extensive rearrangements early in development. During their initial growth, neurones apparently form synaptic connections with an excessive number of targets, later retracting a portion of these synapses in establishing the adult neural circuits. Because of the profound effects which experience has upon the developing nervous system, a question of considerable interest has been the role which the functional use of these developing synapses might play in determining the final pattern of connectivity. At the neuromuscular junction the early changes in synaptic connections are well documented, and here questions about the importance of function can be relatively easily addressed. Mammalian skeletal muscle fibres experience a perinatal period of synapse elimination so that all but one of several synapses formed on each muscle fibre are lost. This synapse elimination is sensitive to alterations of neuromuscular use or activity. Reduction of muscle use by tenotomy or by paralysis of the muscle with drugs blocking nerve impulse conduction or neuromuscular transmission delays or even prevents synapse loss, while increased use produced by stimulation of the muscle nerve apparently accelerates the rate at which synapses are lost. I report here a further examination of the role of neuromuscular activity in synapse elimination. I show that chronic neuromuscular stimulation accelerates synapse elimination but that this acceleration is dependent on the temporal pattern in which the stimuli are presented: brief stimulus trains containing 100 Hz bursts of stimuli produce this acceleration whereas the same number of stimuli presented continuously at 1 Hz do not. Furthermore, the 100 Hz activity pattern which is effective in altering synapse elimination also alters two other muscle properties: the sensitivity of the muscle fibers to acetylcholine and the 'speed' of muscle contractions. These findings suggest that the ability of muscle fibres to maintain more than one nerve terminal, like other muscle properties, is sensitive to the pattern of muscle use rather than just the total amount of use.     

Elimination of action potentials blocks the structural development of retinogeniculate synapses

Although the influence of electrical activity on neural development has been studied extensively, experiments have only recently focused on the role of activity in the development of the mammalian central nervous system (CNS). Using tetrodotoxin (TTX) to abolish sodium-mediated action potentials, studies on the visual system show that impulse activity is essential both for the normal development of neuronal size and responsivity in the lateral geniculate nucleus (LGN), and for the eye-specific segregation of geniculo-cortical axons. There have been no anatomical studies to investigate the influence of action potentials on CNS synaptic development. We report here the first direct evidence that elimination of action potentials in the mammalian CNS blocks the growth of developing axon terminals and the formation of normal adult synaptic patterns. Our results show that when TTX is used to eliminate retinal ganglion-cell action potentials in the cat from birth to 8 weeks, the connections made by ganglion cell axons with LGN neurones, retinogeniculate synapses, remain almost identical morphologically to those in the newborn kitten.     

东方簇盾吸虫神经系统的研究

应用乙酰胆碱酯酶定位方法对东方簇盾吸虫(Lophotaspis orientalis Faust and Tang,1936)成虫的神经系统进行了研究.成虫的神经系统十分复杂,包括中枢神经结、脑神经联合、纵行的神经干、横向的神经连合.中枢神经结位于咽前部两侧,有1粗大的脑神经联合连接.在脑神经联合的下方有1个小的环形的结构.中枢神经结向前发出1对前背神经干和1对前侧神经干,这2对神经干向前的分支汇入围口腔的神经环.中枢神经结向后发出1对后背神经干和1对后腹神经干.后背神经干之间有横向的神经相连,在体末端与后腹神经干相汇合.后腹神经干最为粗大,并在虫体末端汇合,它在腹盘处有2对分支进入腹盘,前1对与腹盘边缘神经相连,后1对贯穿腹盘,其分支构成腹吸盘内复杂的神经网.腹吸盘的指状突起内也有神经分支.     

Changes in the dendritic branching of adult mammalian neurones revealed by repeated imaging in situ

A major obstacle to understanding the mechanism of long-term change in the vertebrate nervous system has been the inability to observe the same nerve cell at different times during the life of an animal. The possibility that changes in neural connectivity underlie the remarkable flexibility of the nervous systems of mammals has therefore not been tested by direct observation. Here, we report studies in which we have visualized the same neurone in the superior cervical ganglion of young adult mice at intervals of up to 33 days. This collection of nerve cells is particularly accessible and therefore well suited to our approach. We find that the dendritic branches of the neurones examined change appreciably over intervals of 2 weeks or more; some branches retract, others elongate and others seem to form de novo. The apparent remodelling of these postsynaptic elements implies that the synaptic connections of these cells normally undergo significant rearrangement beyond what is usually considered to be the developmental period.     

Parallel colour-opponent pathways to primary visual cortex

The trichromatic primate retina parses the colour content of a visual scene into 'red/green' and 'blue/yellow' representations. Cortical circuits must combine the information encoded in these colour-opponent signals to reconstruct the full range of perceived colours. Red/green and blue/yellow inputs are relayed by the lateral geniculate nucleus (LGN) of thalamus to primary visual cortex (V1), so understanding how cortical circuits transform these signals requires understanding how LGN inputs to V1 are organized. Here we report direct recordings from LGN afferent axons in muscimol-inactivated V1. We found that blue/yellow afferents terminated exclusively in superficial cortical layers 3B and 4A, whereas red/green afferents were encountered only in deeper cortex, in lower layer 4C. We also describe a distinct cortical target for 'blue-OFF' cells, whose afferents terminated in layer 4A and seemed patchy in organization. The more common 'blue-ON' afferents were found in 4A as well as lower layer 2/3. Chromatic information is thus conveyed to V1 by parallel, anatomically segregated colour-opponent systems, to be combined at a later stage of the colour circuit.     

Segregation of efferent connections and receptive field properties in visual area V2 of the macaque

V2 is a visual area of the macaque monkey which is at the second level in a recently proposed hierarchy of cortical visual areas. Histochemical staining for cytochrome oxidase (CO) in V2 reveals a pattern of alternate thick and thin CO-rich stripes separated by CO-sparse interstripes. These subregions receive distinct inputs from neurones in CO-rich and CO-sparse zones arrayed within the superficial layers of V1 (refs 4, 5). Are output projections from V2 to higher visual areas also segregated? Using an anatomical double-label paradigm, we have now demonstrated that V2 cells projecting to two of its major target areas, MT and V4 (refs 6, 7), are arranged in stripe-like clusters which are largely segregated from one another and which are closely related to the pattern of CO stripes. Concomitant electrophysiological recordings from V2 indicate that groups of cells having similar receptive field properties are clustered within the subregions defined by these anatomical techniques.     

Effect of impulse blockage on cytochrome oxidase activity in monkey visual system

Cytochrome oxidase (cytochrome c oxidase; ferrocytochrome c: oxygen oxidoreductase, EC 1.9.2.1) has been introduced as an oxidative metabolic marker for neurones in the central nervous system. Previous studies have shown that mature neurones remained sensitive to altered functional demands, and that both developing and adult neurones responded to sensory deprivation or deafferentation by reducing their cytochrome oxidase (Cyt. Ox.) activity. More recently, we showed that the blockage of retinal impulse transmission with tetrodotoxin led to a reversible reduction in Cyt. Ox. staining of affected lateral geniculate (LGN) and striate neurones in adult cats. The present study sought to extend these findings to adult monkeys, where Cyt. Ox. 'puffs' or 'blobs' are uniquely present in the visual cortex. We found that, while the retina remained histologically intact, with only moderate decreases in Cyt. Ox. staining of large ganglion cells and the two plexiform layers, subtle changes occurred in the LGN as early as 1 day post-tetrodotoxin injection, and clear reduction in enzyme levels was evident in both the LGN and the visual cortex by 3 days. Changes became progressively more severe up to 4 weeks post-injection. Within area 17, alternating bands of high and low Cyt. Ox. staining occurred in lamina IV, with alternating rows of dark and lightly reactive puffs superimposed in exact register. Thus, the mature visual neurones in the primate remain extremely sensitive to the cessation of retinal impulse transmission, and plastic metabolic changes occur through several synapses along the sensory pathway.     

Retinotopic order appears before ocular separation in developing visual pathways

In mammals, the major subcortical visual structures receive projections from both eyes, with the uncrossed projection being smaller than the crossed. Each projection is arranged as a separate orderly map of one hemiretina. Although these hemiretinal maps are separate in the nuclei, they are aligned so that the representations of points in the visual field are in register, thus there is a continuity of visual field representation between them. During the early development of the binocular pathways, terminals from the two eyes overlap almost entirely. As development proceeds, terminals arising from each eye segregate to form the adult pattern. In the present study, local retinal lesions were made in ferrets at various stages in development before the separation of the projections from the two eyes. A neuronal tracer was then injected into the damaged eye, defining the pattern of projection from that eye. As reported here, the lesion resulted in a limited interruption in the pattern of terminal label on both sides of the brain, demonstrating that terminals from each eye are arranged in an orderly retinotopic manner at this stage. hence, during later development, as one projection is reduced relative to the other, the two maps must slide in relation to each other.     

Order in the initial retinotectal map in Xenopus: a new technique for labelling growing nerve fibres

Retinal nerve fibres form an orderly map of visual space in several centres in the vertebrate brain. Such topographic maps are a common feature of central nervous system organization, yet the way in which they develop is poorly understood. Early nerve projections in the fetal and neonatal mammalian brain have been found in several cases to be less restricted than those in the adult, suggesting that nerve fibres may initially form a diffuse set of connections in their target structure from which the adult map is sculpted by the elimination of terminals. Indeed, previous electrophysiological data indicate that the retinotectal map in Xenopus laevis might be initially disorganized. We report here, however, that the retinotectal projection is ordered from the beginning of tectal innervation (stage 39/40). We demonstrate this first autoradiographically by tracing groups of growing ganglion cell axons which we labelled by incubating sectors of eye rudiments, before axonal outgrowth, in 3H-proline and replacing them orthotopically. Separate labelling of dorsal and ventral parts of the initial projection showed that retinal fibres are organized topographically, as in the adult, in the tectal rudiment and throughout much of the pathway. Second, we show that visual responses are ordered in the tectum from the first stage that they can be mapped (stage 40). We conclude that the topographic ordering of retinotectal connections develops as a result of directed axonal outgrowth.     

Moving visual stimuli rapidly induce direction sensitivity of developing tectal neurons

During development of the visual system, the pattern of visual inputs may have an instructive role in refining developing neural circuits. How visual inputs of specific spatiotemporal patterns shape the circuit development remains largely unknown. We report here that, in the developing Xenopus retinotectal system, the receptive field of tectal neurons can be 'trained' to become direction-sensitive within minutes after repetitive exposure of the retina to moving bars in a particular direction. The induction of direction-sensitivity depends on the speed of the moving bar, can not be induced by random visual stimuli, and is accompanied by an asymmetric modification of the tectal neuron's receptive field. Furthermore, such training-induced changes require spiking of the tectal neuron and activation of a NMDA (N-methyl-D-aspartate) subtype of glutamate receptors during training, and are attributable to an activity-induced enhancement of glutamate-mediated inputs. Thus, developing neural circuits can be modified rapidly and specifically by visual inputs of defined spatiotemporal patterns, in a manner consistent with predictions based on spike-time-dependent synaptic modification.     

Normal maturation involves systematic changes in binocular visual connections in Xenopus laevis

Systematic changes in neuronal connections have been observed during the development of many vertebrate neuronal systems. These changes have usually involved a refinement from an initial exuberance of connections or a response to some experimental perturbation. Here we report on a system of neuronal connections, which, during a protracted developmental period, undergo ordered changes in response to normally occurring changes in functional requirements. In the frog Xenopus laevis, interocular alignment changes markedly during late larval and post-metamorphic life, producing a progressive enlargement of the binocular portion of the visual field. An intertectal system links the two mid-brain optic tecta and is concerned with the neural representation of binocular visual space. In the adult animal, connections in this system link corresponding points (points receiving information from one locus of binocular visual space) on the two tecta. Changes in eye position with development, however, change the set of corresponding points. Therefore, if the intertectal connections link corresponding tectal points throughout development, they must undergo an ordered change with time. We present electrophysiological evidence that the intertectal connections do, indeed, undergo such changes in response to changes in eye alignment, and that the changes are major.     

Genetic evidence that relative synaptic efficacy biases the outcome of synaptic competition

Synaptic activity drives synaptic rearrangement in the vertebrate nervous system; indeed, this appears to be a main way in which experience shapes neural connectivity. One rearrangement that occurs in many parts of the nervous system during early postnatal life is a competitive process called 'synapse elimination'. At the neuromuscular junction, where synapse elimination has been analysed in detail, muscle fibres are initially innervated by multiple axons, then all but one are withdrawn and the 'winner' enlarges. In support of the idea that synapse elimination is activity dependent, it is slowed or speeded when total neuromuscular activity is decreased or increased, respectively. However, most hypotheses about synaptic rearrangement postulate that change depends less on total activity than on the relative activity of the competitors. Intuitively, it seems that the input best able to excite its postsynaptic target would be most likely to win the competition, but some theories and results make other predictions. Here we use a genetic method to selectively inhibit neurotransmission from one of two inputs to a single target cell. We show that more powerful inputs are strongly favoured competitors during synapse elimination.     

Neurogenesis in the adult is involved in the formation of trace memories

The vertebrate brain continues to produce new neurons throughout life. In the rat hippocampus, several thousand are produced each day, many of which die within weeks. Associative learning can enhance their survival; however, until now it was unknown whether new neurons are involved in memory formation. Here we show that a substantial reduction in the number of newly generated neurons in the adult rat impairs hippocampal-dependent trace conditioning, a task in which an animal must associate stimuli that are separated in time. A similar reduction did not affect learning when the same stimuli are not separated in time, a task that is hippocampal-independent. The reduction in neurogenesis did not induce death of mature hippocampal neurons or permanently alter neurophysiological properties of the CA1 region, such as long-term potentiation. Moreover, recovery of cell production was associated with the ability to acquire trace memories. These results indicate that newly generated neurons in the adult are not only affected by the formation of a hippocampal-dependent memory, but also participate in it.     

Neurite arborization and mosaic spacing in the mouse retina require DSCAM

To establish functional circuitry, retinal neurons occupy spatial domains by arborizing their processes, which requires the self-avoidance of neurites from an individual cell, and by spacing their cell bodies, which requires positioning the soma and establishing a zone within which other cells of the same type are excluded. The mosaic patterns of distinct cell types form independently and overlap. The cues that direct these processes in the vertebrate retina are not known. Here we show that some types of retinal amacrine cells from mice with a spontaneous mutation in Down syndrome cell adhesion molecule (Dscam), a gene encoding an immunoglobulin-superfamily member adhesion molecule, have defects in the arborization of processes and in the spacing of cell bodies. In the mutant retina, cells that would normally express Dscam have hyperfasciculated processes, preventing them from creating an orderly arbor. Also, their cell bodies are randomly distributed or pulled into clumps rather than being regularly spaced mosaics. Our results indicate that mouse DSCAM mediates isoneuronal self-avoidance for arborization and heteroneuronal self-avoidance within specific cell types to prevent fasciculation and to preserve mosaic spacing. These functions are analogous to those of Drosophila DSCAM (ref. 6) and DSCAM2 (ref. 7). DSCAM may function similarly in other regions of the mammalian nervous system, and this role may extend to other members of the mammalian Dscam gene family.     

Electrical activity in early neuronal development

The construction of the brain during embryonic development was thought to be largely independent of its electrical activity. In this view, proliferation, migration and differentiation of neurons are driven entirely by genetic programs and activity is important only at later stages in refinement of connections. However, recent findings demonstrate that activity plays essential roles in early development of the nervous system. Activity has similar roles in the incorporation of newly born neurons in the adult nervous system, suggesting that there are general rules underlying activity-dependent development. The extensive involvement of activity makes it likely that it is required at all developmental stages as a necessary partner with genetic programs.     

Receptive fields in the body-surface map in adult cortex defined by temporally correlated inputs

Receptive fields (RFs) obtained at specific cortical sites can be used to define a topographic map of the body surface in adult mammalian somatosensory cortex. This map is not static, and RFs at particular cortical sites can change in size and location throughout adult life. Conversely, the cortical loci at which a given skin surface is represented can shift hundreds of micrometres across the cortex in the koniocortical field, area 3b (refs 1-12). This plasticity suggests that RFs derive not from rigid anatomical connections, but by the selection of a subset of a large number of inputs. We have proposed that inputs are selected on the basis of temporal correlation 11-15. Here we test this idea by altering the correlation of inputs from two adjacent digits on the adult owl monkey hand by surgically connecting the skin surfaces of the two fingers (the formation of syndactyly). This manipulation increases the correlation of inputs from skin surfaces of adjacent fingers. The striking discontinuity between the zones of representation of adjacent digits on the somatosensory cortex disappeared. These results support the hypothesis that the topography of the body-surface map in the adult cortex is influenced by the temporal correlations of afferent inputs.