Dendritic Branch Formation Research Articles

Insights From Graph Theory on the Morphologies of Actomyosin Networks with Multilinkers

Quantifying the influence of microscopic details on the dynamics of development of the overall structure of a filamentous network is important in a number of biologically relevant contexts, but it is not obvious what order parameters can be used to adequately describe this complex process. In this paper we investigated the role of multivalent actin-binding proteins (ABPs) in reorganizing actin filaments into higher-order complex networks via a computer model of semiflexible filaments. We characterize the importance of local connectivity among actin filaments, as well as the global features of actomyosin networks. We first map the networks into local graph representations and then, using principles from network-theory order parameters, combine properties from these representations to gain insight into the heterogeneous morphologies of actomyosin networks at a global level. We find that ABPs with a valency greater than 2 promote filament bundles and large filament clusters to a much greater extent than bivalent multilinkers. We also show that active myosinlike motor proteins promote the formation of dendritic branches from a stalk of actin bundles. Our work motivates future studies to embrace network theory as a tool to characterize complex morphologies of actomyosin detected by experiments, leading to a quantitative understanding of the role of ABPs in manipulating the self-assembly of actin filaments into unique architectures that underlie the structural scaffold of a cell relating to its mobility and shape.

Insights from graph theory on the morphologies of actomyosin networks with multilinkers.

Quantifying the influence of microscopic details on the dynamics of development of the overall structure of a filamentous network is important in a number of biologically relevant contexts, but it is not obvious what order parameters can be used to adequately describe this complex process. In this paper we investigated the role of multivalent actin-binding proteins (ABPs) in reorganizing actin filaments into higher-order complex networks via a computer model of semiflexible filaments. We characterize the importance of local connectivity among actin filaments, as well as the global features of actomyosin networks. We first map the networks into local graph representations and then, using principles from network-theory order parameters, combine properties from these representations to gain insight into the heterogeneous morphologies of actomyosin networks at a global level. We find that ABPs with a valency greater than 2 promote filament bundles and large filament clusters to a much greater extent than bivalent multilinkers. We also show that active myosinlike motor proteins promote the formation of dendritic branches from a stalk of actin bundles. Our work motivates future studies to embrace network theory as a tool to characterize complex morphologies of actomyosin detected by experiments, leading to a quantitative understanding of the role of ABPs in manipulating the self-assembly of actin filaments into unique architectures that underlie the structural scaffold of a cell relating to its mobility and shape.

Transient lamellipodia predict sites of dendritic branch formation in hippocampal neurons.

Extensive branching creates the complex dendritic arbor of mammalian CNS neurons but capturing the complete process of branch formation with time-lapse recordings has been challenging. Here, we report that application of BMP7 to cultured hippocampal neurons accelerated dendritic growth sufficiently to document branches forming in less than 20h via frequent time-lapse imaging (10-min intervals). In these recordings, most branches emerged as collateral sprouts from the shaft of a parent branch. Analysis of the recordings showed that filopodia were abundant and formed transiently throughout the length of dendrites but among these, only a small subset occurred at sites where branches later emerged. Conversely, formation of lamellipodia was rare and coincided with sites where collateral branches emerged. This pattern suggests that lamellipodial structures act as an important intermediate form of cytoskeletal remodeling related to a cellular commitment to branch, whereas filopodia appear to be related to events prior to such commitment.

Recent advances in branching mechanisms underlying neuronal morphogenesis

Proper neuronal wiring is central to all bodily functions, sensory perception, cognition, memory, and learning. Establishment of a functional neuronal circuit is a highly regulated and dynamic process involving axonal and dendritic branching and navigation toward appropriate targets and connection partners. This intricate circuitry includes axo-dendritic synapse formation, synaptic connections formed with effector cells, and extensive dendritic arborization that function to receive and transmit mechanical and chemical sensory inputs. Such complexity is primarily achieved by extensive axonal and dendritic branch formation and pruning. Fundamental to neuronal branching are cytoskeletal dynamics and plasma membrane expansion, both of which are regulated via numerous extracellular and intracellular signaling mechanisms and molecules. This review focuses on recent advances in understanding the biology of neuronal branching.

Perturbation of canonical and non-canonical BMP signaling affects migration, polarity and dendritogenesis of mouse cortical neurons.

Bone morphogenetic protein (BMP) signaling has been implicated in the regulation of patterning of the forebrain and as a regulator of neurogenesis and gliogenesis in the mammalian cortex. However, its role in other aspects of cortical development in vivo remains unexplored. We hypothesized that BMP signaling might regulate additional processes during the development of cortical neurons after observing active BMP signaling in a spatiotemporally dynamic pattern in the mouse cortex. Our investigation revealed that BMP signaling specifically regulates the migration, polarity and the dendritic morphology of upper layer cortical neurons born at E15.5. On further dissection of the role of canonical and non-canonical BMP signaling in each of these processes, we found that migration of these neurons is regulated by both pathways. Their polarity, however, appears to be affected more strongly by canonical BMP signaling, whereas dendritic branch formation appears to be somewhat more strongly affected by LIMK-mediated non-canonical BMP signaling.

Voltage-gated calcium and sodium channels mediate Sema3A retrograde signaling that regulates dendritic development

Growing axons rely on local signaling at the growth cone for guidance cues. Semaphorin3A (Sema3A), a secreted repulsive axon guidance molecule, regulates synapse maturation and dendritic branching. We previously showed that local Sema3A signaling in the growth cones elicits retrograde retrograde signaling via PlexinA4 (PlexA4), one component of the Sema3A receptor, thereby regulating dendritic localization of AMPA receptor GluA2 and proper dendritic development. In present study, we found that nimodipine (voltage-gated L-type Ca2+ channel blocker) and tetrodotoxin (TTX; voltage-gated Na+ channel blocker) suppress Sema3A-induced dendritic localization of GluA2 and dendritic branch formation in cultured hippocampal neurons. The local application of nimodipine or TTX to distal axons suppresses retrograde transport of Venus-Sema3A that has been exogenously applied to the distal axons. Sema3A facilitates axonal transport of PlexA4, which is also suppressed in neurons treated with either TTX or nimodipine. These data suggest that voltage-gated calcium and sodium channels mediate Sema3A retrograde signaling that regulates dendritic GluA2 localization and branch formation.

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.

Microtubule minus-end binding protein CAMSAP2 controls axon specification and dendrite development.

In neurons, most microtubules are not associated with a central microtubule-organizing center (MTOC), and therefore, both the minus and plus-ends of these non-centrosomal microtubules are found throughout the cell. Microtubule plus-ends are well established as dynamic regulatory sites in numerous processes, but the role of microtubule minus-ends has remained poorly understood. Using live-cell imaging, high-resolution microscopy, and laser-based microsurgery techniques, we show that the CAMSAP/Nezha/Patronin family protein CAMSAP2 specifically localizes to non-centrosomal microtubule minus-ends and is required for proper microtubule organization in neurons. CAMSAP2 stabilizes non-centrosomal microtubules and is required for neuronal polarity, axon specification, and dendritic branch formation in vitro and in vivo. Furthermore, we found that non-centrosomal microtubules in dendrites are largely generated by γ-Tubulin-dependent nucleation. We propose a two-step model in which γ-Tubulin initiates the formation of non-centrosomal microtubules and CAMSAP2 stabilizes the free microtubule minus-ends in order to control neuronal polarity and development.

Golden goal controls dendrite elongation and branching of multidendritic arborization neurons in Drosophila

Precise refinement of axonal and dendritic patterns is essential for the maturation of functional neuronal circuits. Although several transmembrane molecules have been shown to control the development of both axons and dendrites, the molecular mechanisms that regulate these different processes are poorly understood. Golden Goal (Gogo) is one of the molecules that are known to control the development of axons in the Drosophila visual system. In this study, we analyzed Gogo function in dendritic field formation of dorsal multidendritic arborization (md-da) neurons of the Drosophila Peripheral Nervous System. We showed that Gogo is required to restrain the growth of ddaC dendrites toward the midline in the embryo. During larval stages, Gogo promotes dendritic branching of the complex classIV ddaC neurons. However, over-expression of Gogo restrained dendritic branch formation in ddaC neurons, and this phenotype was enhanced by co-over-expression with Flamingo (Fmi), a partner of Gogo in axon guidance. These results suggest Gogo plays important roles in maintaining homeostasis of dendritic branching. Like axons, the cytoplasmic part of Gogo is required for its function in dendritic tree formation, suggesting that Gogo conveys information from extracellular cues to intracellular molecules that control dendrite development.

Inositol 1,4,5-Trisphosphate Receptor-Mediated Calcium Release in Purkinje Cells: From Molecular Mechanism to Behavior

The inositol 1,4,5-trisphosphate (IP(3)) receptor is highly expressed in cerebellar Purkinje cells and mediates conspicuous calcium release from intracellular calcium stores. Receptor stimulation, such as through mGluR1, activates the G(q)-PLC pathway, which leads to IP(3)-induced calcium release and subsequent cellular responses, including cerebellar long-term depression in Purkinje cells. Recent studies have demonstrated the regulatory mechanisms of IP(3) receptor, revealing activation via IP(3) and Ca(2+), inactivation via high concentrations of Ca(2+), and modulation by various proteins that bind to the IP(3) receptor. Novel calcium imaging techniques and caged compounds provide analysis of calcium signals at the single spine level in relation to the induction of long-term depression. Genetically encoded indicators for calcium or IP(3) could provide alternate Ca(2+) or IP(3) imaging, in particular, for in vivo observations. IP(3)-induced calcium release participates in early development of dendritic branch formation, and loss-of-function mutations or hyper-activation could result various diseases. The IP(3) receptor plays a central role in calcium signaling in Purkinje cells, affecting a wide variety of cellular functions, including development, plasticity, maintenance of synaptic functions, and cerebellar motor control.

C. elegans bicd-1, homolog of theDrosophiladynein accessory factorBicaudal D, regulates the branching of PVD sensory neuron dendrites

The establishment of cell type-specific dendritic arborization patterns is a key phase in the assembly of neuronal circuitry that facilitates the integration and processing of synaptic and sensory input. Although studies in Drosophila and vertebrate systems have identified a variety of factors that regulate dendrite branch formation, the molecular mechanisms that control this process remain poorly defined. Here, we introduce the use of the Caenorhabditis elegans PVD neurons, a pair of putative nociceptors that elaborate complex dendritic arbors, as a tractable model for conducting high-throughput RNAi screens aimed at identifying key regulators of dendritic branch formation. By carrying out two separate RNAi screens, a small-scale candidate-based screen and a large-scale screen of the ~3000 genes on chromosome IV, we retrieved 11 genes that either promote or suppress the formation of PVD-associated dendrites. We present a detailed functional characterization of one of the genes, bicd-1, which encodes a microtubule-associated protein previously shown to modulate the transport of mRNAs and organelles in a variety of organisms. Specifically, we describe a novel role for bicd-1 in regulating dendrite branch formation and show that bicd-1 is likely to be expressed, and primarily required, in PVD neurons to control dendritic branching. We also present evidence that bicd-1 operates in a conserved pathway with dhc-1 and unc-116, components of the dynein minus-end-directed and kinesin-1 plus-end-directed microtubule-based motor complexes, respectively, and interacts genetically with the repulsive guidance receptor unc-5.

Differential Roles of NMDA Receptor Subtypes NR2A and NR2B in Dendritic Branch Development and Requirement of RasGRF1

N-methyl-D-aspartate receptors (NMDARs) are known to regulate axonal refinement and dendritic branching. However, because NMDARs are abundantly present as tri-heteromers (e.g., NR1/NR2A/NR2B) during development, the precise role of the individual subunits NR2A and NR2B in these processes has not been elucidated. Ventral spinal cord neurons (VSCNs) provide a unique opportunity to address this problem, because the expression of both NR2A and NR2B (but not NR1) is downregulated in culture. Exogenous NR2A or NR2B were introduced into these naturally NR2-null neurons at 4 DIV, and electrophysiological recordings at 11 DIV confirmed that synaptic NR1NR2A receptors and NR1NR2B receptors were formed, respectively. Analysis of the dendritic architecture showed that introduction of NR2B, but not NR2A, dramatically increased the number of secondary and tertiary dendritic branches of VSCNs. Whole cell patch-clamp recordings further indicated that the newly formed branches in NR2B-expressing neurons were able to establish functional synapses because the frequency of miniature AMPA-receptor synaptic currents was increased. Using previously described mutants, we also found that disruption of the interaction between NR2B and RasGRF1 dramatically impaired dendritic branch formation in VSCNs. The differential role of the NR2A and NR2B subunits and the requirement for RasGRF1 in regulating branch formation was corroborated in hippocampal cultures. We conclude that the association between NR1NR2B-receptors and RasGRF1 is needed for dendritic branch formation in VSCNs and hippocampal neurons in vitro. The dominated NR2A expression and the limited interactions of this subunit with the signaling protein RasGRF1 may contribute to the restricted dendritic arbor development in the adult CNS.

Dendrite Elongation and Dendritic Branching Are Affected Separately by Different Forms of Intrinsic Motoneuron Excitability

Dendrites are the fundamental determinant of neuronal wiring. Consequently dendritic defects are associated with numerous neurological diseases and mental retardation. Neuronal activity can have profound effects on dendritic structure, but the mechanisms controlling distinct aspects of dendritic architecture are not fully understood. We use the Drosophila genetic model system to test the effects of altered intrinsic excitability on postembryonic dendritic architecture development. Targeted dominant negative knock-downs of potassium channel subunits allow for selectively increasing the intrinsic excitability of a selected subset of motoneurons, whereas targeted expression of a genetically modified noninactivating potassium channel decrease intrinsic excitability in vivo. Both manipulations cause significant dendritic overgrowth, but by different mechanisms. Increased excitability causes increased dendritic branch formation, whereas decreased excitability causes increased dendritic branch elongation. Therefore dendritic branching and branch elongation are controlled by separate mechanisms that can be addressed selectively in vivo by different manipulations of neuronal intrinsic excitability.

Ancient rules of memory

The first kiss, the smell of mother's cooking or the vision of a beautiful landscape; memories such as these are etched firmly into our brains for recall in vivid detail, even decades later. Memories and experience influence our decisions throughout life, both consciously and subconsciously—in a sense, our memories make us what we are. Yet, understanding how information is encoded in our brains and how memories are maintained, both at the conceptual and molecular levels, remains one of the greatest challenges in the life sciences. Through decades of research, biologists have answered many fundamental questions about the brain, and more recent work has revealed the identity of several crucial molecules involved in the storage and retrieval of information. This has at last begun to shed light on the major scientific puzzle: how do we form and maintain long‐term memories? ![][1] The field of neurobiology had made relatively little progress toward understanding memory since the Canadian psychologist Donald Hebb (1904–1985) introduced the idea of synaptic plasticity nearly 60 years ago (Hebb, 1949). Hebb explained the establishment of memory by changes in the number and strength of synaptic connections between neurons. According to his theory, synapses respond to stimulation by neurotransmitters in a cumulative manner, and repeated stimulation—for example, by learning—leads to ever‐stronger connections between specific neurons until they become permanent. Over time, neuroscientists accepted Hebb's idea of synaptic plasticity, despite a lack of direct molecular evidence. In fact, it was not until recently that scientists—in large part Eric Kandel at Columbia University in New York, NY, USA—discovered several biochemical mechanisms that now support Hebb's idea. Kandel himself won the Nobel Prize for medicine in 2000 for his discovery that the syntheses of different proteins relate to short‐term and long‐term memory in the sea slug Aplysia . The foundations for this work and … [1]: /embed/graphic-1.gif

The origin recognition core complex regulates dendrite and spine development in postmitotic neurons

The origin recognition complex (ORC) ensures exactly one round of genome replication per cell cycle through acting as a molecular switch that precisely controls the assembly, firing, and inactivation of the replication initiation machinery. Recent data indicate that it may also coordinate the processes of mitosis and cytokinesis and ensure the proper distribution of replicated genome to daughter cells. We have found that the ORC core subunits are highly expressed in the nervous system. They are selectively localized to the neuronal somatodendritic compartment and enriched in the membrane fraction. siRNA knockdown of ORC subunits dramatically reduced dendritic branch formation and severely impeded dendritic spine emergence. Expression of ORC ATPase motif mutants enhanced the branching of dendritic arbors. The ORC core complex thus appears to have a novel role in regulating dendrite and dendritic spine development in postmitotic neurons.

Rac1 and Fragile X Syndrome

The most common form of inherited mental retardation is associated with loss-of-function (LOF) mutations in the fragile X mental retardation 1 ( FMR1 ) gene. Although FMR1 encodes an RNA-binding protein that associates with mRNAs and ribosomes, its molecular function and role in nervous system development have not been clear. The homologous protein in Drosophila , Fmr1, is highly expressed in dendritic arborization (DA) neurons of the peripheral nervous system. Lee et al. studied DA neurons from Fmr1-deficient flies and determined that the protein regulates dendritic branching during development. Fmr1 localized to the cytoplasm and dendrites of DA neurons. Fmr1 LOF mutant embryos exhibited increased dendritic processes, whereas embryos overexpressing Fmr1 had reduced dendritic branch formation. mRNA encoding the small guanosine triphosphatase Rac1 immunoprecipitated with Fmr1 from larval lysates. DA neurons in larva expressing a null allele of Rac1 developed fewer dendritic branches than wild-type neurons, similar to the phenotype observed when Fmr1 was overexpressed. Rac1 overexpression promoted dendritic branching and partially rescued the effects of Fmr1 overexpression, indicating that Rac1 is a downstream component of Fmr1 function. Because Rac1 has been implicated in controlling cell morphology, the authors propose that local regulation of Rac1 expression by Fmr1 in neurons may control dendritic branching during development. A. Lee, W. Li, K. Xu, B. A. Bogert, K. Su, F.-B. Gao, Control of dendritic development by the Drosophila fragile X-related gene involves the small GTPase Rac1. Development 130 , 5543-5552 (2003). [Abstract] [Full Text]

Rac1 and Fragile X Syndrome

The most common form of inherited mental retardation is associated with loss-of-function (LOF) mutations in the fragile X mental retardation 1 (FMR1) gene. Although FMR1 encodes an RNA-binding protein that associates with mRNAs and ribosomes, its molecular function and role in nervous system development have not been clear. The homologous protein in Drosophila, Fmr1, is highly expressed in dendritic arborization (DA) neurons of the peripheral nervous system. Lee et al. studied DA neurons from Fmr1-deficient flies and determined that the protein regulates dendritic branching during development. Fmr1 localized to the cytoplasm and dendrites of DA neurons. Fmr1 LOF mutant embryos exhibited increased dendritic processes, whereas embryos overexpressing Fmr1 had reduced dendritic branch formation. mRNA encoding the small guanosine triphosphatase Rac1 immunoprecipitated with Fmr1 from larval lysates. DA neurons in larva expressing a null allele of Rac1 developed fewer dendritic branches than wild-type neurons, similar to the phenotype observed when Fmr1 was overexpressed. Rac1 overexpression promoted dendritic branching and partially rescued the effects of Fmr1 overexpression, indicating that Rac1 is a downstream component of Fmr1 function. Because Rac1 has been implicated in controlling cell morphology, the authors propose that local regulation of Rac1 expression by Fmr1 in neurons may control dendritic branching during development.A. Lee, W. Li, K. Xu, B. A. Bogert, K. Su, F.-B. Gao, Control of dendritic development by the Drosophila fragile X-related gene involves the small GTPase Rac1. Development 130, 5543-5552 (2003). [Abstract] [Full Text]

Calcium-dependent alterations in dendritic architecture of hippocampal pyramidal neurons.

Dendritic arbor formation and the underlying mechanisms are crucial for the functional connectivity and plasticity of neurons. We used a focal electric field to locally raise calcium levels in individual dendritic shafts of isolated hippocampal pyramidal neurons, in order to develop an accessible system for studying dendritic branch formation, and to test the role of calcium as an intrinsic signal that may participate in arborization. Filopodia were induced in a manner temporally and spatially related to induced calcium rises. Certain filopodia also thickened and were transformed into dendritic branches. These results suggest that calcium-mediated signaling can induce branching in dendrites, and describe an accessible system for studying the intracellular machinery that drives dendritic arborization.