HGF regulates the development of cortical pyramidal dendrites

Although hepatocyte growth factor (HGF) and its receptor tyrosine kinase MET are widely expressed in the developing and mature central nervous system, little is known about the role of MET signaling in the brain. We have used particle-mediated gene transfer in cortical organotypic slice cultures established from early postnatal mice to study the effects of HGF on the development of dendritic arbors of pyramidal neurons. Compared with untreated control cultures, exogenous HGF promoted a highly significant increase in dendritic growth and branching of layer 2 pyramidal neurons, whereas inactivation of endogenous HGF with function-blocking, anti-HGF antibody caused a marked reduction in size and complexity of the dendritic arbors of these neurons. Furthermore, pyramidal neurons transfected with an MET dominant-negative mutant receptor likewise had much smaller and less complex dendritic arbors than did control transfected neurons. Our results indicate that HGF plays a role in regulating dendritic morphology in the developing cerebral cortex.     

Sex differences in the motor nucleus of cranial nerve IX-X in Xenopus laevis: a quantitative Golgi study

In the clawed frog (Xenopus laevis), motor neurons in cranial nerve nucleus IX-X control contraction of laryngeal muscles responsible for sexually dimorphic vocal behaviors. We examined sex differences in dendritic arbors of n.IX-X cells using the Golgi-Cox method. Three morphological classes of somal types (ovoid, triangular, and elongate) are present in similar frequencies in n.IX-X of both males and females. The male n.IX-X neuron is a more complex and hypertrophied version of the female n.IX-X cell. The number of primary dendrites is the same for both sexes, but males have more total dendritic segments. The overall dendritic length of male n.IX-X neurons is two to three times that of the female. Males have longer dendritic segments between all branch points. Male and female frogs differ in levels of circulating androgens; neurons of n.IX-X are targets for androgenic steroids. To determine if androgen can affect dendritic morphology in adult females, we examined Golgi-impregnated cells in n.IX-X from ovariectomized females treated with testosterone for 1 month. The total number of dendritic segments was reduced by androgen treatment due to reduction in the number of higher order dendritic segments; the number of primary dendritic segments was unchanged. Androgen treatment may induce resorption of higher order dendritic branches. The overall dendritic length of androgen-treated female n.IX-X neurons was unchanged, and dendritic segments were longer. Thus, although androgen can alter dendrites of n.IX-X cells in adult females, this short-term treatment does not produce a masculine dendritic architecture.     

Control of axonal sprouting and dendrite branching by the Nrg-Ank complex at the neuron-glia interface

Neurons are highly polarized cells with distinct subcellular compartments, including dendritic arbors and an axon. The proper function of the nervous system relies not only on correct targeting of axons, but also on development of neuronal-class-specific geometry of dendritic arbors [1-4]. To study the intercellular control of the shaping of dendritic trees in vivo, we searched for cell-surface proteins expressed by Drosophila dendritic arborization (da) neurons [5-7]. One of them was Neuroglian (Nrg), a member of the Ig superfamily ; Nrg and vertebrate L1-family molecules have been implicated in various aspects of neuronal wiring, such as axon guidance, axonal myelination, and synapse formation [9-12]. A subset of the da neurons in nrg mutant embryos exhibited deformed dendritic arbors and abnormal axonal sprouting. Our functional analysis in a cell-type-selective manner strongly suggested that those da neurons employed Nrg to interact with the peripheral glia for suppressing axonal sprouting and for forming second-order dendritic branches. At least for the former role, Nrg functioned in concert with the intracellular adaptor protein Ankyrin (Ank) [13]. Thus, the neuron-glia interaction that is mediated by Nrg, together with Ank under some situations, contributes to axonal and dendritic morphogenesis.     

Dendritic pattern development of the honeybee antennal lobe neurons: a laser scanning confocal microscopic study.

The processing of odorant signals is performed, in the olfactory bulb of vertebrates or in the antennal lobe of insects, by different types of neurons which display specific morphological and functional features. The present work characterizes the morphogenesis of the main neuronal types which participate in olfactory discrimination in the adult honeybee (Apis mellifera). Neurons were stained intracellularly with Lucifer yellow at different stages of pupal development and in the adult, and imaged by laser scanning confocal microscopy. Attending to branching patterns, all pupal neurons could be attributed to morphological types previously established in the adult. Given the functional importance of intraglomerular dendritic arbors in the processing of olfactory information, the study focused on their development. The two main classes, dense and sparse intraglomerular arbors, display adultlike features as early as the second day of pupal development. However, morphometric measurements and confocal observations show that their general pattern undergoes continuous maturation processes until late pupal stages and after emergence of the adult. Among these, the results point out a pruning of dendritic branches in sparse arbors, but not in dense arbors.     

Degeneration of Axotomized Projection Neurons in the Rat dLGN: Temporal Progression of Events and Their Mitigation by a Single Administration of FGF2

Removal of visual cortex in the rat axotomizes projection neurons in the dorsal lateral geniculate nucleus (dLGN), leading to cytological and structural changes and apoptosis. Biotinylated dextran amine was injected into the visual cortex to label dLGN projection neurons retrogradely prior to removing the cortex in order to quantify the changes in the dendritic morphology of these neurons that precede cell death. At 12 hours after axotomy we observed a loss of appendages and the formation of varicosities in the dendrites of projection neurons. During the next 7 days, the total number of dendrites and the cross-sectional areas of the dendritic arbors of projection neurons declined to about 40% and 20% of normal, respectively. The response of dLGN projection neurons to axotomy was asynchronous, but the sequence of structural changes in individual neurons was similar; namely, disruption of dendrites began within hours followed by cell soma atrophy and nuclear condensation that commenced after the loss of secondary dendrites had occurred. However, a single administration of fibroblast growth factor-2 (FGF2), which mitigates injury-induced neuronal cell death in the dLGN when given at the time of axotomy, markedly reduced the dendritic degeneration of projection neurons. At 3 and 7 days after axotomy the number of surviving dendrites of dLGN projection neurons in FGF-2 treated rats was approximately 50% greater than in untreated rats, and the cross-sectional areas of dendritic arbors were approximately 60% and 50% larger. Caspase-3 activity in axotomized dLGN projection neurons was determined by immunostaining for fractin (fractin-IR), an actin cleavage product produced exclusively by activated caspase-3. Fractin-IR was seen in some dLGN projection neurons at 36 hours survival, and it increased slightly by 3 days. A marked increase in reactivity was seen by 7 days, with the entire dLGN filled with dense fractin-IR in neuronal cell somas and dendrites.     

Ectopic Purkinje cells in the cerebellar white matter of normal adult rodents: a Golgi study

In Golgi/Río-Hortega preparations of rat and rabbit cerebellar vermis we have occasionally found isolated ectopic Purkinje cells in the white matter. They were located beneath the bases of the folia and their dendritic branches extended within the confines of the white matter without penetrating into the overlying cortical layers. The general morphology of these ectopic cells was variable, particularly in the extension and shape of the dendritic trees, but all of them exhibited a lower density of dendritic branches than normal Purkinje cells. The less-developed ectopic neurons had multipolar dendritic trees with nonplanar branches irregularly studded with spines. The well-developed ones displayed a more extensive arborization of their processes and they usually preserved some morphological features of normal cortical Purkinje cells: distal dendritic branches studded with numerous spines, a pear-shaped soma, clearly defined morphological polarity and a tendency to display planar arrangement of the dendritic arbors. In semithin sections these neurons also showed cytological features of normal Purkinje cells, such as the Nissl substance forming a nuclear cap oriented toward the dendritic pole. We suggest that the abnormal location of the neurons results from a disorder of Purkinje cell migration which occurs naturally during the prenatal development of the cerebellum. The possible morphogenetic mechanisms involved in the migration and differentiation of these ectopic neurons are also discussed.     

The Proprotein Convertase KPC-1/Furin Controls Branching and Self-avoidance of Sensory Dendrites in Caenorhabditis elegans

Animals sample their environment through sensory neurons with often elaborately branched endings named dendritic arbors. In a genetic screen for genes involved in the development of the highly arborized somatosensory PVD neuron in C. elegans, we have identified mutations in kpc-1, which encodes the homolog of the proprotein convertase furin. We show that kpc-1/furin is necessary to promote the formation of higher order dendritic branches in PVD and to ensure self-avoidance of sister branches, but is likely not required during maintenance of dendritic arbors. A reporter for kpc-1/furin is expressed in neurons (including PVD) and kpc-1/furin can function cell-autonomously in PVD neurons to control patterning of dendritic arbors. Moreover, we show that kpc-1/furin also regulates the development of other neurons in all major neuronal classes in C. elegans, including aspects of branching and extension of neurites as well as cell positioning. Our data suggest that these developmental functions require proteolytic activity of KPC-1/furin. Recently, the skin-derived MNR-1/menorin and the neural cell adhesion molecule SAX-7/L1CAM have been shown to act as a tripartite complex with the leucine rich transmembrane receptor DMA-1 on PVD mechanosensory to orchestrate the patterning of dendritic branches. Genetic analyses show that kpc-1/furin functions in a pathway with MNR-1/menorin, SAX-7/L1CAM and DMA-1 to control dendritic branch formation and extension of PVD neurons. We propose that KPC-1/furin acts in concert with the ‘menorin’ pathway to control branching and growth of somatosensory dendrites in PVD.     

Development of morphological diversity of dendrites in Drosophila by the BTB-zinc finger protein abrupt

Morphological diversity of dendrites contributes to specialized functions of individual neurons. In the present study, we examined the molecular basis that generates distinct morphological classes of Drosophila dendritic arborization (da) neurons. da neurons are classified into classes I to IV in order of increasing territory size and/or branching complexity. We found that Abrupt (Ab), a BTB-zinc finger protein, is expressed selectively in class I cells. Misexpression of ab in neurons of other classes directed them to take the appearance of cells with smaller and/or less elaborated arbors. Loss of ab functions in class I neurons resulted in malformation of their typical comb-like arbor patterns and generation of supernumerary branch terminals. Together with the results of monitoring dendritic dynamics of ab-misexpressing cells or ab mutant ones, all of the data suggested that Ab endows characteristics of dendritic morphogenesis of the class I neurons.     

NMDA receptor activation induces translocation and activation of Rac in mouse hippocampal area CA1

Neuronal development requires several discrete morphological steps that are believed to involve the small GTPase Rac. For example, neural activity, through NMDA receptors and/or AMPA receptors, activates Rac leading to elaboration of dendritic arbors. In the current study, we have conducted studies which indicate that Rac might be an important molecule involved in morphological plasticity in the adult mouse. We demonstrate that Rac is expressed at synapses in the adult mouse hippocampus. We also demonstrate that treatment of hippocampal slices with NMDA induces membrane translocation and activation of Rac in area CA1. Interestingly, we also find that there is an increase in Rac that is associated with NMDA receptor complexes following NMDA receptor activation. Taken together, our data are consistent with the idea that Rac could be participating in NMDA receptor-dependent changes in morphology that occur during synaptic plasticity and memory formation in the adult mouse hippocampus.     

Dendritic morphology is altered in hippocampal neurons following prenatal compromise

Chronic placental insufficiency (CPI), a known cause of intrauterine growth restriction, can lead to structural alterations in the developing brain that might underlie postnatal neurological deficits. We have previously demonstrated significant reductions in the volumes of hippocampal neuropil layers in fetal guinea pig brains following experimentally induced growth restriction. To determine the components of the neuropil affected in the brains of growth restricted (GR) fetuses, the dendritic morphology of CA1 pyramidal neurons and dentate granule cells was examined. CPI was induced by unilateral uterine artery ligation in pregnant guinea pigs at midgestation (term approximately 67 days). Hippocampi from control and GR fetuses were stained using the Rapid Golgi technique and the growth and branching of the dendritic arbors were quantified using the Sholl method. In addition, the density of dendritic spines was determined on the apical arbors of each population. In GR brains (n = 7) compared to controls (n = 7), there was a reduction in dendritic elongation (p < 0.005) and an alteration in the branch point distribution in CA1 basal arbors, and a reduction both in the outgrowth (p < 0.05) and branch point number (p < 0.05) of CA1 apical arbors. Dentate granule cells from GR brains also demonstrated reduced dendritic outgrowth (p < 0.05). There was an increase in dendritic spine density in both neuronal populations; this might be due either to altered synaptic pruning or as a compensatory mechanism for reduced dendritic length. These findings demonstrate that a chronic prenatal insult causes selective changes in the morphology of hippocampal cell dendrites and may lead to alterations in hippocampal function in the postnatal period.     

Chronic stress alters dendritic morphology in rat medial prefrontal cortex

Chronic stress produces deficits in cognition accompanied by alterations in neural chemistry and morphology. Medial prefrontal cortex is a target for glucocorticoids involved in the stress response. We have previously demonstrated that 3 weeks of daily corticosterone injections result in dendritic reorganization in pyramidal neurons in layer II-III of medial prefrontal cortex. To determine if similar morphological changes occur in response to chronic stress, we assessed the effects of daily restraint stress on dendritic morphology in medial prefrontal cortex. Male rats were exposed to either 3 h of restraint stress daily for 3 weeks or left unhandled except for weighing during this period. On the last day of restraint, animals were overdosed and brains were stained using a Golgi-Cox procedure. Pyramidal neurons in lamina II-III of medial prefrontal cortex were drawn in three dimensions, and the morphology of apical and basilar arbors was quantified. Sholl analyses demonstrated a significant alteration of apical dendrites in stressed animals: overall, the number and length of apical dendritic branches was reduced by 18 and 32%, respectively. The reduction in apical dendritic arbor was restricted to distal and higher-order branches, and may reflect atrophy of terminal branches: terminal branch number and length were reduced by 19 and 35%. On the other hand, basilar dendrites were not affected. This pattern of dendritic reorganization is similar to that seen after daily corticosterone injections. This reorganization likely reflects functional changes in prefrontal cortex and may contribute to stress-induced changes in cognition.     

Beta-catenin stabilization extends regulatory T cell survival and induces anergy in nonregulatory T cells

Beta-catenin is a central molecule in the Wnt pathway. Expression of a stable form of beta-catenin on CD4+CD25+ regulatory T (T(reg)) cells resulted in a marked enhancement of survival of these cells in vitro. Furthermore, stable beta-catenin-expressing CD4+CD25+ T(reg) cells outcompeted control T(reg) cells in vivo, and the number of T(reg) cells necessary for protection against inflammatory bowel disease could be substantially reduced when stable beta-catenin-expressing CD4+CD25+ T(reg) cells were used instead of control T(reg) cells. Expression of stable beta-catenin on potentially pathogenic CD4+CD25- T cells rendered these cells anergic, and the beta-catenin-mediated induction of anergy occurred even in Foxp3-deficient T cells. Thus, through enhanced survival of existing regulatory T cells, and through induction of unresponsiveness in precursors of T effector cells, beta-catenin stabilization has a powerful effect on the prevention of inflammatory disease.     

大鼠视皮层神经元电生理和形态学特性在发育中的变化

为研究大鼠不同发育阶段视皮层神经元电的生理学与形态学特性,实验观察了神经元电生理和形态学特性的变化与年龄的同步化程度,探讨视皮层视觉依赖性突触的形成和重新分布的细胞内机制。应用脑片膜片钳全细胞记录技术和细胞内生物家标记相结合的方法,记录4—28d SD大鼠视皮层神经元的突触后电流(postsynaptic currents,PSCs)。共记录156个大鼠视皮层神经元,睁眼前与睁眼后组中无反应型细胞数量,多突触反应型细胞数量、细胞的输入阻抗有显著性差异。成功标记23例神经元,不同年龄的神经元的形态学成熟度不同。低输入阻抗神经元在形态学上属成熟型,高输入阻抗神经元属幼稚型。该结果表明,大鼠在发育过程中,视皮层神经元功能的成熟表现为在形觉刺激以及局部神经元网络的整合作用下的视觉依赖性突触的形成和重新分布。在视觉发育可塑性关键期内,视皮层神经元形态和电生理特性的变化与年龄的同步化程度大于皮层下结构。     

Regulation of dendritic maintenance and growth by a mammalian 7-pass transmembrane cadherin

Drosophila Flamingo is a 7-pass transmembrane cadherin that is necessary for dendritic patterning and axon guidance. How it works at the molecular level and whether homologs of Flamingo play similar roles in mammalian neurons or not have been unanswered questions. Here, we performed loss-of-function analysis using an RNAi system and organotypic brain slice cultures to address the role of a mammalian Flamingo homolog, Celsr2. Knocking down Celsr2 resulted in prominent simplification of dendritic arbors of cortical pyramidal neurons and Purkinje neurons, and this phenotype seemed to be due to branch retraction. Cadherin domain-mediated homophilic interaction appears to be required for the maintenance of dendritic branches. Furthermore, expression of various Celsr2 forms elicited distinct responses that were dependent on an extracellular subregion outside the cadherin domains and on a portion within the carboxyl intracellular tail. Based on these findings, we discuss how Celsr2 may regulate dendritic maintenance and growth.     

Dendritic pattern development of the honeybee antennal lobe neurons: a laser scanning confocal microscopic study

The processing of odorant signals is performed, in the olfactory bulb of vertebrates or in the antennal lobe of insects, by different types of neurons which display specific morphological and functional features. The present work characterizes the morphogenesis of the main neuronal types which participate in olfactory discrimination in the adult honeybee (Apis mellifera). Neurons were stained intracellularly with Lucifer yellow at different stages of pupal development and in the adult, and imaged by laser scanning confocal microscopy. Attending to branching patterns, all pupal neurons could be attributed to morphological types previously established in the adult. Given the functional importance of intraglomerular dendritic arbors in the processing of olfactory information, the study focused on their development. The two main classes, dense and sparse intraglomerular arbors, display adultlike features as early as the second day of pupal development. However, morphometric measurements and confocal observations show that their general pattern undergoes continuous maturation processes until late pupal stages and after emergence of the adult. Among these, the results point out a pruning of dendritic branches in sparse arbors, but not in dense arbors. © 1999 John Wiley & Sons, Inc. J Neurobiol 39: 461–474, 1999     

Quantitative analysis of electrical properties of dendritic spines

Several sugestions have been made with regard to the functional significance of dendritic spines in connection with synaptic plasticity. We have shown that for a constant synaptic current, when the synaptic resistance is large compared to the spine-stem resistance, a morphological change in the spine does not produce a marked change in the postsynaptic potential (PSP). When the synaptic resistance is comparable to the spine-stem impedance a morphological change in the spine can induce changes in the synaptic current and the PSP due to the so-called nonlinear effect to the synapse (Kawato and Tsukahara, 1983, 1984). Consequently, in a study of the electrical properties of dendritic spines the input impedance of the parent dendrite, the spinestalk conductance and the conductance change associated with synaptic activity must be considered. We quantitatively estimated all three factors. By comparing electrophysiological data with morphological data, we estimated the synaptic conductance which causes corticorubral EPSP. Its maximum amplitude was 43 nS with a time-to-peak value of 0.3 ms. With this value, the effects of the spine were examined using an improved algorithm based on that of Butz and Cowan (1974). It uses a three-dimensional morphology of the rubrospinal (RS) neurons, which was reconstructed from serial sections containing HRP-filled RS cells. As the spine shortens, the amplitude of the EPSP becomes considerably larger, but its time-to-peak value does not markedly change. Moreover, if unitary EPSP in the RS cell is produced by the activation of several synaptic terminals a morphological change of the spine has a smaller effect on the EPSPs.     

Changes in the structural complexity of the aged brain

Structural changes of neurons in the brain during aging are complex and not well understood. Neurons have significant homeostatic control of essential brain functions, including synaptic excitability, gene expression, and metabolic regulation. Any deviations from the norm can have severe consequences as seen in aging and injury. In this review, we present some of the structural adaptations that neurons undergo throughout normal and pathological aging and discuss their effects on electrophysiological properties and cognition. During aging, it is evident that neurons undergo morphological changes such as a reduction in the complexity of dendrite arborization and dendritic length. Spine numbers are also decreased, and because spines are the major sites for excitatory synapses, changes in their numbers could reflect a change in synaptic densities. This idea has been supported by studies that demonstrate a decrease in the overall frequency of spontaneous glutamate receptor-mediated excitatory responses, as well as a decrease in the levels of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid and N-methyl-d-aspartate receptor expression. Other properties such as gamma-aminobutyric acid A receptor-mediated inhibitory responses and action potential firing rates are both significantly increased with age. These findings suggest that age-related neuronal dysfunction, which must underlie observed decline in cognitive function, probably involves a host of other subtle changes within the cortex that could include alterations in receptors, loss of dendrites, and spines and myelin dystrophy, as well as the alterations in synaptic transmission. Together these multiple alterations in the brain may constitute the substrate for age-related loss of cognitive function.     

Cell type-specific dendritic polarity in the absence of spatially organized external cues

Pyramidal neurons of the hippocampus and cortex have polarized dendritic arbors, but little is known about the cellular mechanisms distinguishing apical and basal dendrites. We used morphometric analysis and time lapse imaging of cultured hippocampal neurons to show that glutamatergic neurons develop progressive dendritic asymmetry in the absence of polarized extrinsic cues. Thus, pyramidal neurons have a cellular program for polarized dendrite growth independent of tissue microenvironment.     

Glycinergic transmission regulates dendrite size in organotypic culture

We previously demonstrated that inhibitory synaptic transmission influences dendrite development in vivo. We now report an analogous finding in an organotypic culture of a glycinergic projection nucleus, the medial nucleus of the trapezoid body (MNTB), and its postsynaptic target, the lateral superior olive (LSO) of gerbils. Cultures were generated at 6–7 days postnatal and grown in serum containing medium with or without the glycine receptor antagonist, strychnine (SN), at 2 μM. LSO neurons were then labeled with biocytin, and the dendritic arbors were analyzed morphometrically. Compared to neurons from age-matched in vivo tissue, the neurons cultured in control media were somewhat atrophic, including decreases in dendritic branching and length. Incubation in strychnine led to a dramatic increase in dendritic branching and total dendritic length. Control neurons averaged 6.3 branches, compared to 18 branches/neuron in SN-treated cultures. There was a similar increase in primary dendrites and total dendritic length. The physical elimination of MNTB cells did not mimic SN treatment, presumably because glycinergic LSO neurons generated intrinsic connections. In fact, the LSO soma area was significantly greater following MNTB removal, suggesting that these afferents provide a second signal to postsynaptic neurons. These results suggest that spontaneous glycinergic transmission regulates the growth of postsynaptic processes. © 1996 John Wiley & Sons, Inc.