Visual afferentation deficit during diffuse photosensitivity period in nestlings affects morphogenesis in Wulst neurons

Golgi-stained preparations were processed with computerized morphometry to study the effect of the deprivation (eyes covered with nontransparent caps) on the development of neurons in the Wulst (the structure analogous to mammalian visual cortex) of pied flycatcher nestlings. Six-day-old nestlings that have only one form of the visual perception--diffuse photosensitivity--were studied. In the previous paper [Korneeva et al., 1994] in was shown that the Wulst of 6-day-old nestlings consisted of non-differentiated neurons and stellate-like cells at different stages of maturation; the latter group was subdivided into more- and less-mature cells. This work proved that even a 1.5-day-long deprivation (actual duration being counted off from the moment of appearance of the first retinal photoreceptors at the age of 4.5 days) resulted in significant changes in the geometry of stellate-like neurons. The changes in less-mature cells were predominantly destructive (decline of all quantitative indices of a cell, including the significant decrease in the total length of dendrites, maximal radius of the dendritic field and cell branching index), while the changes in more-mature cells were constructive (increase in all quantitative cell indices, including statistically significant increase in the soma section area, total length of dendrites, maximal radius of dendritic field and the number of foci of maximal branching). Different reactions of these cell types to the limitation of visual afferentation may be connected with differences in the afferent inflow to less- and more-mature cells or/and with different maturational stages of these neurons at the onset of deprivation.     

The morphogenesis of Wulst neurons in the pied flycatcher under conditions of limited afferent inflow]

Golgi preparations of the pied flycatcher Wulst region (the structure analogous to the mammalian visual cortex) were analyzed using the method of computerized morphometry, to study the influence of visual deprivation on the development of different types of neurons selected previously. Deprivation was accomplished by covering the young's eye with nontransparent caps. The experiments were carried out in 10-day-old nestlings (the onset of patterned vision) and 13-day-old nestlings (functioning patterned vision). In 10-day-old nestlings, the deprivation produced constructive changes in dendritic apparatus of projective stellate cells (among them, the most pronounced was more than three-fold increase in the number of foci of maximal branching) practically not affecting the small stellate-like cells. In 13-day-old nestlings, cells belonging to all selected cell types underwent destructive changes: their quantitative characteristics were decreased as compared to those in control nestlings. A large number of tree-like neurons were revealed in the Wulst in the deprived 10-day-old nestlings while in the control age-matched nestlings they were virtually never found. This phenomenon may be explained by the increased affinity to impregnation evoked by deprivation-induced biochemical changes in the tree-like neurons or to increase in their number. In the latter case, the phenomenon may be considered as compensatory, directed at the establishing of contacts with nonvisual afferents.     

Relais cells and afferent axons in the dorsal part of the lateral geniculate body in the albino rat under geometrical aspects]

1. The average volumes of dendritic domaines of relay neurons (P-neurons) were calculated and the quantitative relations to the neuronal elements situated in this area were investigated. Likewise we carried out measurements and calculations at the terminal parts of afferent axons, to find a conception concerning possible contacts between axons and P-neurons considering quantitative aspects. 2. The dendrites of one P-neuron are distributed in an area of about 0,008 mum3. In this area there are located somata of at least 120 other P-neurons and dendrites of altogether about 900 P-neurons. 3. The type-1-axons (cortical afferents) run almost linearly in the longitudinal system of the CGLd. Traversing a distance adequate to the diameter of a P-neuron (250 mum) the dendrites of 150 to 170 P-neurons may cross the course of one axon. At this distance the axon, however, has just set up about 50 boutons, thus synaptic contacts may be established with one third at most of the existing cells. A type-1-axon that is bifurcating in the entrance area into the CGLd is altogether of about 2000 mum in length and is able to develop about 420 presynaptic profiles. 4. The type-2-axons (retinal afferents) show a distinct terminal branching zone. The Golgi-Kopsch impregnated terminals of type-2a-axons are distributed in a space of 147000 mum3 capacity, the corresponding terminals of type-2b-axons in a space of 443000 mum3. The type-2a-axons having an average number of 23 boutons, may contact the dendritic branching zones of 25 P-neurons. There is a good reason to assume that type-2b-axons are in contact also with terminal dendritic parts of P-neurons. Thus the number of P-cells, which spread their dendrites into the terminal branching zone of one type-2b-axon may amount to 540. The average number of boutons of one type-2b-terminal, however, is only about 160. This means that synaptic contacts may be developed to the P-neurons-dendrites not exceeding 30% of them. 5. Various aspects of divergence of axon terminals in the albino rat's CGLd are discussed.     

A Golgi study of the isthmic nuclei in the pigeon (Columba Iivia)

Summary The isthmic nuclei of the pigeon were studied with the use of three different Golgi techniques. The nucleus isthmo-opticus (IO) consists of a single cell type in which all dendrites of one neuron take the same direction and ramify at identical distances from the perikaryon to form dense dendritic arborizations. The cell bodies of the IO neurons form two parallel layers. The dendrites of these neurons always extend to the area between the two layers so that the dendritic arborizations of opposite neurons overlap. A model of the cellular organization of the IO was constructed based upon these morphological characteristics. The neurons of the n. isthmi/pars parvocellularis (Ipc) have oval perikarya and long, smooth, infrequently branching dendrites. All neurons except those at the borders of the nucleus show the same dorsoventral orientation in their dendritic arborizations and together with their afferents seem to have a columnar organization. The dendrites of the neurons located at the margin of the nucleus ramify within the Ipc along its border. The n. semilunaris (Slu) consists of neurons with round somata that have on an average three dendrites with small spines. The axons leave the nucleus from the medial side and join the lemniscus lateralis. The neurons of the n. isthmi/pars magnocellularis (Imc) comprise a generalized isodendritic type resembling the cells of the reticular formation. Axons from the tectum penetrate the nucleus, making numerous en-passant contacts with several neurons.     

Specific reduction of development of the Mauthner neuron lateral dendrite after otic capsule ablation in Brachydanio rerio

During development, afferent fibers may stimulate development of postsynaptic target neurons. By surgically ablating an otic vesicle in zebrafish embryos 30 hr after fertilization we deprived the developing Mauthner (M) neuron of vestibular axonal input to its lateral dendrite. After 8 days, 14 M cells were examined by light microscopy, and in each case the size and branching of the lateral dendrite was reduced. No consistent changes were observed in shape and size of other regions of the deprived cells or in the contralateral control cells. Synapses onto five of these pairs of cells were examined by electron microscopy. Except for missing vestibular terminals on the deprived dendrites, the synaptic input to the dendrites and to other regions of the M cell was normal in distribution and pattern. These data suggest that growth-promoting or trophic effects of vestibular axons upon the M cell are localized to its lateral dendrite.     

Nature of the distribution of synapses in the neurons of the reticular formation and some afferent nuclei]

Under study was the morphology of synaptic terminations of the brain reticular formation in cats and dogs as well as that of the afferent nuclei of the cat's posterior columns. The neurons of the Goll's and Burdach's nuclei have a richly ramified dendritic network. In this connection the main mass of synapses is disposed on the dendrites and their different branchings. The dendrites of the multipolar cells of the reticular formation have the main type of branching, but the cells are distributed from the nerve cell body at a considerable distance (up to 50 mu and more). The synapses are observed at the total length of the dendrite, so the axo-dendritic contacts quantitatively prevail over axo-somatic ones. The morphological data are compared with physiological axo-somatic and axo-dendritic concepts of the role of synapses in conducting the nerve impulse.     

The Morphological Identity of Insect Dendrites

Dendrite morphology, a neuron's anatomical fingerprint, is a neuroscientist's asset in unveiling organizational principles in the brain. However, the genetic program encoding the morphological identity of a single dendrite remains a mystery. In order to obtain a formal understanding of dendritic branching, we studied distributions of morphological parameters in a group of four individually identifiable neurons of the fly visual system. We found that parameters relating to the branching topology were similar throughout all cells. Only parameters relating to the area covered by the dendrite were cell type specific. With these areas, artificial dendrites were grown based on optimization principles minimizing the amount of wiring and maximizing synaptic democracy. Although the same branching rule was used for all cells, this yielded dendritic structures virtually indistinguishable from their real counterparts. From these principles we derived a fully-automated model-based neuron reconstruction procedure validating the artificial branching rule. In conclusion, we suggest that the genetic program implementing neuronal branching could be constant in all cells whereas the one responsible for the dendrite spanning field should be cell specific.     

Polarized secretory trafficking directs cargo for asymmetric dendrite growth and morphogenesis

Proper growth of dendrites is critical to the formation of neuronal circuits, but the cellular machinery that directs the addition of membrane components to generate dendritic architecture remains obscure. Here, we demonstrate that post-Golgi membrane trafficking is polarized toward longer dendrites of hippocampal pyramidal neurons in vitro and toward apical dendrites in vivo. Small Golgi outposts partition selectively into longer dendrites and are excluded from axons. In dendrites, Golgi outposts concentrate at branchpoints where they engage in post-Golgi trafficking. Within the cell body, the Golgi apparatus orients toward the longest dendrite, and this Golgi polarity precedes asymmetric dendrite growth. Manipulations that selectively block post-Golgi trafficking halt dendrite growth in developing neurons and cause a shrinkage of dendrites in mature pyramidal neurons. Further, disruption of Golgi polarity produces neurons with symmetric dendritic arbors lacking a single longest principal dendrite. These results define a novel polarized organization of neuronal secretory trafficking and demonstrate a mechanistic link between directed membrane trafficking and asymmetric dendrite growth.     

Development of the Drosophila mushroom bodies: elaboration,remodeling and spatial organization of dendrites in the calyx

One Drosophila mushroom body (MB) is derived from four indistinguishable cell lineages, development of which involves sequential generation of multiple distinct types of neurons. Differential labeling of distinct MB clones reveals that MB dendrites of different clonal origins are well mixed at the larval stage but become restricted to distinct spaces in adults. Interestingly, a small dendritic domain in the adult MB calyx remains as a fourfold structure that, similar to the entire larval calyx, receives dendritic inputs from all four MB clones. Mosaic analysis of single neurons demonstrates that MB neurons, which are born around pupal formation, acquire unique dendritic branching patterns and consistently project their primary dendrites into the fourfold dendritic domain. Distinct dendrite distribution patterns are also observed for other subtypes of MB neurons. In addition, pruning of larval dendrites during metamorphosis allows for establishment of adult-specific dendrite elaboration/distribution patterns. Taken together, subregional differences exist in the adult Drosophila MB calyx, where processing and integration of distinct types of sensory information begin.     

Different levels of the homeodomain protein cut regulate distinct dendrite branching patterns of Drosophila multidendritic neurons

Functionally similar neurons can share common dendrite morphology, but how different neurons are directed into similar forms is not understood. Here, we show in embryonic and larval development that the level of Cut immunoreactivity in individual dendritic arborization (da) sensory neurons correlates with distinct patterns of terminal dendrites: high Cut in neurons with extensive unbranched terminal protrusions (dendritic spikes), medium levels in neurons with expansive and complex arbors, and low or nondetectable Cut in neurons with simple dendrites. Loss of Cut reduced dendrite growth and class-specific terminal branching, whereas overexpression of Cut or a mammalian homolog in lower-level neurons resulted in transformations toward the branch morphology of high-Cut neurons. Thus, different levels of a homeoprotein can regulate distinct patterns of dendrite branching.     

Interneurons of the crayfish brain: The relationship between dendrite location and afferent input

This study was undertaken to examine the relationship between the structure and function of the descending interneurons of the crayfish brain. In particular, the dendritic fields were examined to ascertain if the location of an interneuronal dendrite in any of the six cerebral hemineuromeres (which subserve specific sensory modalities) is a necessary or sufficient condition to determine the functional and/or synaptic input to the interneuron. If a neuron projects a dendrite to a hemineuromere of the deutocerebrum or tritocerebrum, the neuron derives sensory input from the corresponding afferent root in 95% of our observations. Most of these inputs (86%) contain at the least a monosynaptic component. Conversely, if a cell derives monosynaptic input from any one of three of the four deutocerebral and tritocerebral roots tested, it has a corresponding dendrite (in 98% of our observations) in the appropriate hemineuromere. Input from the contrlateral antennal nerve is an exception to this rule. The presence of a dendrite in the protocerebrum is not sufficient for predicting detectable visual input, but every instance of detectable visual input is associated with a protocerebral dendrite. Polysynaptic inputs are frequently (42%) not associated with corresponding dendrites. In neurons that were repeatedly dye filled in different animals, we observed significant variation only in the number and precise location of the smaller secondary and tertiary neurites. This variation rarely influenced the subset of sensory lobes innervated by the neuron.     

Cell-type specific dendritic contacts between retinal ganglion cells during development.

The extent of a neuron's dendritic field defines the region within which information is processed. The dendritic fields of functionally distinct ON and OFF center retinal ganglion cells (RGCs) form separate mosaics across the retina. Within each mosaic, neighboring dendritic fields overlap by a constant amount, sampling the visual field with the appropriate coverage. Contact-mediated lateral inhibition between neighboring RGCs has long been thought to regulate both the extent and overlap of dendritic fields during development. Here we show that dendro-dendritic contact exists between developing RGCs and occurs in a manner that would regulate the formation of ON and OFF mosaics separately. Dye-filled neighboring ON and OFF ferret alpha RGCs were reconstructed using multiphoton microscopy. At all neonatal ages examined, we observed dendro-dendritic contacts between RGCs of the same sign (ON/ON; OFF/OFF), but never between cells of opposite signs (ON/OFF). Terminal dendrites of one cell often touched a dendrite of its neighbor as they intersected. In some instances, the distal dendrite of one cell formed a fascicle with the proximal process of its neighbor. Alpha cells did not form contacts with neighboring beta cells of the same sign. Together, these observations suggest that dendro-dendritic contact between RGCs is cell-type specific. Dendritic contacts were observed even before the alpha cell arbors were completely stratified, suggesting that cell-cell recognition may take place early in their development. For each cell type, the relative overlap of dendritic fields was constant with age, despite a two-fold increase in field area. We suggest that dendro-dendritic contacts may be sites of intercellular signaling that could regulate local extension of dendrites to maintain the relative overlap of RGCs within a mosaic during development.     

Eliminating afferent impulse activity does not alter the dendritic branching of the amphibian Mauthner cell

In the developing amphibian, the formation of extra vestibular contacts on the Mauthner cell (M-cell) enhances dendritic branching, while deprivation reduces it (Goodman and Model, 1988a). The mechanism underlying the interaction between afferent fibers and developing dendritic branches is not known; neural activity may be an essential component of the stimulating effect. We examined the role of afferent impulse activity in the regulation of M-cell dendritic branching in the axolotl (Ambystoma mexicanum) embryo. M-cells occur as a pair of large, uniquely identifiable neurons in the axolotl medulla. Synapses from the ipsilateral vestibular nerve (nVIII) are restricted to a highly branched region of the M-cell lateral dendrite. We varied the amount of nVIII innervation and eliminated neural activity. First, unilateral transplantation of a vestibular primordium deprived some M-cells of nVIII innervation and superinnervated others. Second, surgical fusion of axolotls to TTX-harboring California newt (Taricha torosa) embryos paralyzed the Ambystoma twin: voltage-sensitive Na+ channel blockade by TTX eliminated action potential propagation. Reconstruction of M-cells in 18 mm larvae revealed that dendritic growth was influenced by in-growing axons even in the absence of incoming impulses: impulse blockade had no effect on the stimulation of dendritic growth by the afferent fibers.     

Fine structural analysis of calcitonin gene-related peptide in the mouse inferior olivary complex

Climbing fiber afferents to the cerebellum, from the inferior olivary complex, have a powerful excitatory effect on Purkinje cells. Changes in the responsiveness of olivary neurons to their afferent inputs, leading to changes in the firing rate or pattern of activation in climbing fibers, have a significant effect on the activation of cerebellar neurons and ultimately on cerebellar function. Several neuropeptides have been localized in both varicosities and cell bodies of the mouse inferior olivary complex, one of which, calcitonin gene related peptide (CGRP), has been shown to modulate the activity of olivary neurons. The purpose of the present study was to investigate the synaptic relationships of CGRP-containing components of the caudal medial accessory olive and the principal olive of adult mice, using immunohistochemistry and electron microscopy. The vast majority of immunoreactive profiles were dendrites and dendritic spines within and outside the glial boundaries of synaptic glomeruli (clusters). Both received synaptic inputs from non-CGRP labeled axon terminals. CGRP was also present within the somata of olivary neurons as well as in profiles that had cytological characteristics of axons, some of which were filled with synaptic vesicles. These swellings infrequently formed synaptic contacts. At the LM level, few, if any, CGRP-immunoreactive climbing fibers, were seen, suggesting that CGRP is compartmentalized within the somata and dendrites of olivary neurons and is not transported to their axon terminals. Thus, in addition to previously identified extrinsic sources of CGRP, the widespread distribution of CGRP within olivary somata and dendrites identifies an intrinsic source of the peptide suggesting the possibility of dendritic release and a subsequent autocrine or paracrine function for this peptide within olivary circuits.     

Statistical evaluation of dendritic growth models

A mathematical model (Kliemann, W. 1987.Bull. math. Biol. 49, 135–152.) that predicts the quantitative branching pattern of dendritic tree was evaluated using the apical and basal dendrites of rat hippocampal neurons. The Wald statistics for χ²-test was developed for the branching pattern of dendritic trees and for the distribution of the maximal order of the tree. Using this statistic, we obtained a reasonable, but not excellent, fit of the mathematical model for the dendritic data. The model's predictability of branching patterns was greatly enhanced by replacing one of the assumptions used for the original model “splitting of branches for all dendritic orders is stochastically independent”, with a new assumption “branches are more likely to split in areas where there is already a high density of branches”. The modified model delivered an excellent fit for basal dendrites and for the apical dendrites of hippocampal neurons from young rats (30–34 days postpartum). This indicates that for these cells the development of dendritic patterns is the result of a purely random and a systematic component, where the latter one depends on the density of dendritic branches in the brain area considered. For apical dendrites there is a trend towards decreasing pattern predictability with increasing age. This appears to reflect the late arrival of afferents and subsequent synaptogenesis proximal on the apical dendritic tree of hippocampal neurons.     

Cholinergic afferents to gonadotropin-releasing hormone neurons of the rat

Gonadotropin-releasing hormone-synthesizing neurons represent the final common pathway in the hypothalamic regulation of reproduction and their secretory activity is influenced by a variety of neurotransmitters and neuromodulators acting centrally in synaptic afferents to gonadotropin-releasing hormone neurons. The present study examined the anatomical relationship of cholinergic neuronal pathways and gonadotropin-releasing hormone neurons of the preoptic area. The immunocytochemical detection of choline acetyltransferase or vesicular acetylcholine transporter revealed a fine network of cholinergic fibers in this region. At the light microscopic level, the cholinergic axons formed appositions to the gonadotropin-releasing hormone immunoreactive cell bodies and dendrites. Results of electron microscopic studies confirmed the absence of glial interpositions in many of these neuronal contacts. Classical cholinergic synapses, which belonged to the asymmetric category, were only observed rarely on gonadotropin-releasing hormone neurons. The lack of synaptic density in most contacts corroborates previous observations on the cholinergic system elsewhere in the brain. Further, it suggests a dominantly non-synaptic route also in this cholinergic neuronal communication. This study provides direct neuromorphological evidence for the involvement of the cholinergic system in the afferent neuronal regulation of gonadotropin-releasing hormone neurons. The sources of cholinergic afferents and the receptorial mechanisms underlying this interaction will require further clarification.     

Multiple clusters of release sites formed by individual thalamic afferents onto cortical interneurons ensure reliable transmission

Thalamic afferents supply the cortex with sensory information by contacting both excitatory neurons and inhibitory interneurons. Interestingly, thalamic contacts with interneurons constitute such a powerful synapse that even one afferent can fire interneurons, thereby driving feedforward inhibition. However, the spatial representation of this potent synapse on interneuron dendrites is poorly understood. Using Ca imaging and electron microscopy we show that an individual thalamic afferent forms multiple contacts with the interneuronal proximal dendritic arbor, preferentially near branch points. More contacts are correlated with larger amplitude synaptic responses. Each contact, consisting of a single bouton, can release up to seven vesicles simultaneously, resulting in graded and reliable Ca transients. Computational modeling indicates that the release of multiple vesicles at each contact minimally reduces the efficiency of the thalamic afferent in exciting the interneuron. This strategy preserves the spatial representation of thalamocortical inputs across the dendritic arbor over a wide range of release conditions.     

Microtubule nucleation and organization in dendrites

Dendrite branching is an essential process for building complex nervous systems. It determines the number, distribution and integration of inputs into a neuron, and is regulated to create the diverse dendrite arbor branching patterns characteristic of different neuron types. The microtubule cytoskeleton is critical to provide structure and exert force during dendrite branching. It also supports the functional requirements of dendrites, reflected by differential microtubule architectural organization between neuron types, illustrated here for sensory neurons. Both anterograde and retrograde microtubule polymerization occur within growing dendrites, and recent studies indicate that branching is enhanced by anterograde microtubule polymerization events in nascent branches. The polarities of microtubule polymerization events are regulated by the position and orientation of microtubule nucleation events in the dendrite arbor. Golgi outposts are a primary microtubule nucleation center in dendrites and share common nucleation machinery with the centrosome. In addition, pre-existing dendrite microtubules may act as nucleation sites. We discuss how balancing the activities of distinct nucleation machineries within the growing dendrite can alter microtubule polymerization polarity and dendrite branching, and how regulating this balance can generate neuron type-specific morphologies.     

Testosterone regulates the density of dendritic spines in the male preoptic area

Male-typical behavior is dependent on testosterone. Castrated males gradually stop mating and engaging in sexual behaviors. Castrates treated with testosterone regain motivation and sex behaviors over time. Although this effect is well characterized, the specific mechanisms by which testosterone treatment recovers sexual behaviors remain unknown. The medial preoptic area (MPOA) is a likely site for testosterone's action on copulation. The integrity of the area is essential for the expression of male sex behavior; and the MPOA is densely populated with receptors for gonadal steroids. Moreover testosterone appears to regulate synaptic efficacy in the MPOA. Exposure to sexually relevant stimuli stimulates the MPOA but only in the presence of circulating testosterone. Sites afferent to the area respond to similar exposure independent of the hormonal milieu suggesting that testosterone mediates communication between the MPOA and its afferents. The protracted time course suggests that the effects of steroidal manipulation are mediated by structural changes. The present experiment evaluated this hypothesis by comparing dendritic spine density among Syrian hamsters that were castrated, castrated and treated with testosterone, or were left gonadally intact. Brains were sectioned and stained using the rapid Golgi stain protocol (FD Neurotechnologies, Baltimore), and the spine density, dendrite length, and the number of branches were compared among groups. Intact and testosterone replaced animals had more spines and greater spine density but did not differ in dendrite length and branching from castrated animals. These results suggest that existing dendrites increase the number of spines available for synapse formation but do not extend their dendrites in response to testosterone treatment.     

A novel forward genetic screen for identifying mutations affecting larval neuronal dendrite development in Drosophila melanogaster

Vertebrate and invertebrate dendrites are information-processing compartments that can be found on both central and peripheral neurons. Elucidating the molecular underpinnings of information processing in the nervous system ultimately requires an understanding of the genetic pathways that regulate dendrite formation and maintenance. Despite the importance of dendrite development, few forward genetic approaches have been used to analyze the latest stages of dendrite development, including the formation of F-actin-rich dendritic filopodia or dendritic spines. We developed a forward genetic screen utilizing transgenic Drosophila second instar larvae expressing an actin, green fluorescent protein (GFP) fusion protein (actin::GFP) in subsets of sensory neurons. Utilizing this fluorescent transgenic reporter, we conducted a forward genetic screen of >4000 mutagenized chromosomes bearing lethal mutations that affected multiple aspects of larval dendrite development. We isolated 13 mutations on the X and second chromosomes composing 11 complementation groups affecting dendrite outgrowth/branching, dendritic filopodia formation, or actin::GFP localization within dendrites in vivo. In a fortuitous observation, we observed that the structure of dendritic arborization (da) neuron dendritic filopodia changes in response to a changing environment.