BDNF/TrkB mediates long-distance dendritic growth by activating CREB/PI3K-mTOR-dependent translation in neuronal cell bodies

Mapping Intimacies ◽

10.1101/2020.08.22.262923 ◽

2020 ◽

Author(s):

G Moya-Alvarado ◽

F.C Bronfman

Keyword(s):

Cortical Neurons ◽

Dendritic Growth ◽

Neuronal Cell ◽

Protein Translation ◽

Dendritic Arborization ◽

Neuronal Morphology ◽

Neuronal Cell Body ◽

Long Distance ◽

Bdnf Signaling

ABSTRACTBrain-Derived Neurotrophic Factor (BDNF) is broadly expressed in many circuits of the central nervous system (CNS). It binds TrkB and p75 to trigger different signaling pathways, including ERK1/2 and PI3K-mTOR, to induce dendritic growth and synaptic plasticity. When binding to BDNF, TrkB and p75 are endocytosed to signaling endosomes to continue signaling inside the cell. Whether BDNF/TrkB-p75 signaling endosomes in axons are regulating long-distance signaling in cell bodies to modify neuronal morphology is unknown. Here, we studied the functional role of BDNF signaling endosomes in long-distance regulation of dendritic growth using compartmentalized cultures of rat and mouse cortical neurons derived from p75exonIII knock-out or TrkBF616A knock-in mice. By applying BDNF to distal axons, we showed the capacity of axonal BDNF to increase dendritic arborization in cell bodies. This process depended on TrkB activity, but not p75 expression. In axons, BDNF/TrkB co-localized with Rab5 endosomes and increased active Rab5. Also, dynein was required for BDNF long-distance signaling, consistent with sorting and transport of signaling endosomes. Using neurons derived from TrkBF616A knock-in mice and the 1NM-PP1 inhibitor, we were able to demonstrate that TrkB receptors activated in the axons by BDNF, were required in the neuronal cell body to increase TrkB activity and phosphorylation of CREB. Also, we were able to visualize endosomes containing activated TrkB. PI3K activity was not required in the axons for dynein dependent BDNF responses. However, dendritic arborization induced by axonal BDNF signaling required both nuclear CREB and PI3K activation in cell bodies. Consistently, axonal BDNF increased protein translation in cell bodies and CREB and PI3K and mTOR activity were required for this process. Altogether, these results show that BDNF/TrkB signaling endosomes generated in axons allows long-distance control of dendritic growth coordinating both transcription and protein translation. Our results suggest a role of BDNF-TrkB signaling endosomes wiring circuits in the CNS.

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Form and Function in Cells of the Brain

The Neuron ◽

10.1093/med/9780199773893.003.0002 ◽

2015 ◽

pp. 23-38

Author(s):

Irwin B. Levitan ◽

Leonard K. Kaczmarek

Keyword(s):

Axonal Transport ◽

Molecular Motors ◽

Critical Role ◽

Neuronal Cell ◽

Cell Types ◽

Neuronal Cell Body ◽

Long Distance ◽

Form And Function ◽

And Function ◽

The Brain

This chapter examines unique mechanisms that the neuron has evolved to establish and maintain the form required for its specialized signaling functions. Unlike some other organs, the brain contains a variety of cell types including several classes of glial cells, which play a critical role in the formation of the myelin sheath around axons and may be involved in immune responses, synaptic transmission, and long-distance calcium signaling in the brain. Neurons share many features in common with other cells (including glia), but they are distinguished by their highly asymmetrical shapes. The neuronal cytoskeleton is essential for establishing this cell shape during development and for maintaining it in adulthood. The process of axonal transport moves vesicles and other organelles to regions remote from the neuronal cell body. Proteins such as kinesin and dynein, called molecular motors, make use of the energy released by hydrolysis of ATP to drive axonal transport.

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STXBP5/tomosyn regulates the small RhoA GTPase to control the dendritic stability of neurons and the surface expression of AMPA receptors

10.1101/617845 ◽

2019 ◽

Author(s):

Wenjuan Shen ◽

Michaela B.C. Kilander ◽

Morgan S. Bridi ◽

Jeannine A. Frei ◽

Robert F. Niescier ◽

...

Keyword(s):

Synaptic Transmission ◽

Ampa Receptors ◽

Surface Expression ◽

Autism Spectrum ◽

Dendritic Morphology ◽

Dendritic Arborization ◽

Neuronal Morphology ◽

Rhoa Gtpase ◽

Wd40 Domain

AbstractTomosyn, a protein encoded by syntaxin-1-binding protein 5 (STXBP5) gene, has a well-established presynaptic role in the inhibition of neurotransmitter release and the reduction of synaptic transmission by its conical interaction with the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) machinery. The postsynaptic role of tomosyn in dendritic arborization, spine stability, and trafficking of ionotropic glutamate receptors remains to be elucidated. We used short hairpin RNA (shRNA) to knock down tomosyn in mouse primary neurons to evaluate the postsynaptic cellular function and molecular signaling regulated by tomosyn. Knockdown of tomosyn led to an increase of RhoA GTPase activity accompanied by compromised dendritic arborization, loss of dendritic spines, decreased surface expression of AMPA receptors, and reduced miniature excitatory postsynaptic current (mEPSC) frequency. Inhibiting RhoA signaling was sufficient to rescue the abnormal dendritic morphology and the surface expression of AMPA receptors. The function of tomosyn regulating RhoA is mediated through the N-terminal WD40 motif, where two variants each carrying a single nucleotide mutation in this region, were found in individuals with autism spectrum disorder (ASD). We demonstrated that these variants displayed loss-of-function phenotypes. Unlike the wild-type tomosyn, these two variants failed to restore the reduced dendritic complexity, spine density, as well as decreased surface expression of AMPA receptors in tomosyn knockdown neurons. This study uncovers a critical role of tomosyn, independent of its interaction with the SNARE machinery, in maintaining neuronal function by inhibiting RhoA activity. Further analysis of tomosyn variants also provides a potential mechanism for explaining cellular pathology in ASD.Significance StatementThis study unveils a vital role of tomosyn in the maintenance of neuronal morphology, basal synaptic transmission, and AMPA receptor surface expression that is distinct from its presynaptic role. Tomosyn affects dendritic stability and glutamate receptor trafficking via the regulation of the Rho signaling pathway and this interaction is likely independent of the interaction with the dendritic SNARE complex, such as syntaxin-4. The WD40 domain of tomosyn is necessary to conduct the Rho regulation and two autism-associated variants localized at the WD40 domain perturb this function. The current study reveals a novel molecular link between dendritic stability and synaptic function, which could advance a greater understanding of the cellular pathologies involved in neurodevelopmental disorders, such as ASD.

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Ischemic Cerebral Endothelial Cell–Derived Exosomes Promote Axonal Growth

Stroke ◽

10.1161/strokeaha.120.031728 ◽

2020 ◽

Vol 51 (12) ◽

pp. 3701-3712

Author(s):

Yi Zhang ◽

Yi Qin ◽

Michael Chopp ◽

Chao Li ◽

Amy Kemper ◽

...

Keyword(s):

Cortical Neurons ◽

Neuronal Cell ◽

Axonal Growth ◽

Mature Mirnas ◽

Bioinformatic Analysis ◽

Axonal Outgrowth ◽

Ischemic Brain ◽

Neuronal Homeostasis ◽

Neuronal Cell Bodies

Background and Purpose: Cerebral endothelial cells (CECs) and axons of neurons interact to maintain vascular and neuronal homeostasis and axonal remodeling in normal and ischemic brain, respectively. However, the role of exosomes in the interaction of CECs and axons in brain under normal conditions and after stroke is unknown. Methods: Exosomes were isolated from CECs of nonischemic rats and is chemic rats (nCEC-exos and isCEC-exos), respectively. A multicompartmental cell culture system was used to separate axons from neuronal cell bodies. Results: Axonal application of nCEC-exos promotes axonal growth of cortical neurons, whereas isCEC-exos further enhance axonal growth than nCEC-exos. Ultrastructural analysis revealed that CEC-exos applied into distal axons were internalized by axons and reached to their parent somata. Bioinformatic analysis revealed that both nCEC-exos and isCEC-exos contain abundant mature miRNAs; however, isCEC-exos exhibit more robust elevation of select miRNAs than nCEC-exos. Mechanistically, axonal application of nCEC-exos and isCEC-exos significantly elevated miRNAs and reduced proteins in distal axons and their parent somata that are involved in inhibiting axonal outgrowth. Blockage of axonal transport suppressed isCEC-exo–altered miRNAs and proteins in somata but not in distal axons. Conclusions: nCEC-exos and isCEC-exos facilitate axonal growth by altering miRNAs and their target protein profiles in recipient neurons.

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Dual Role of Herpes Simplex Virus 1 pUS9 in Virus Anterograde Axonal Transport and Final Assembly in Growth Cones in Distal Axons

Journal of Virology ◽

10.1128/jvi.03023-15 ◽

2015 ◽

Vol 90 (5) ◽

pp. 2653-2663 ◽

Cited By ~ 12

Author(s):

Monica Miranda-Saksena ◽

Ross A. Boadle ◽

Russell J. Diefenbach ◽

Anthony L. Cunningham

Keyword(s):

Axonal Transport ◽

Cell Body ◽

Virus Assembly ◽

Neuronal Cell ◽

Growth Cones ◽

Neuronal Cell Body ◽

Anterograde Axonal Transport ◽

Simplex Virus ◽

Hsv 1

ABSTRACTThe herpes simplex virus type 1 (HSV-1) envelope protein pUS9 plays an important role in virus anterograde axonal transport and spread from neuronal axons. In this study, we used both confocal microscopy and transmission electron microscopy (TEM) to examine the role of pUS9 in the anterograde transport and assembly of HSV-1 in the distal axon of human and rat dorsal root ganglion (DRG) neurons using US9 deletion (US9−), repair (US9R), and wild-type (strain F, 17, and KOS) viruses. Using confocal microscopy and single and trichamber culture systems, we observed a reduction but not complete block in the anterograde axonal transport of capsids to distal axons as well as a marked (∼90%) reduction in virus spread from axons to Vero cells with the US9 deletion viruses. Axonal transport of glycoproteins (gC, gD, and gE) was unaffected. Using TEM, there was a marked reduction or absence of enveloped capsids, in varicosities and growth cones, in KOS strain and US9 deletion viruses, respectively. Capsids (40 to 75%) in varicosities and growth cones infected with strain 17, F, and US9 repair viruses were fully enveloped compared to less than 5% of capsids found in distal axons infected with the KOS strain virus (which also lacks pUS9) and still lower (<2%) with the US9 deletion viruses. Hence, there was a secondary defect in virus assembly in distal axons in the absence of pUS9 despite the presence of key envelope proteins. Overall, our study supports a dual role for pUS9, first in anterograde axonal transport and second in virus assembly in growth cones in distal axons.IMPORTANCEHSV-1 has evolved mechanisms for its efficient transport along sensory axons and subsequent spread from axons to epithelial cells after reactivation. In this study, we show that deletion of the envelope protein pUS9 leads to defects in virus transport along axons (partial defect) and in virus assembly and egress from growth cones (marked defect). Virus assembly and exit in the neuronal cell body are not impaired in the absence of pUS9. Thus, our findings indicate that pUS9 contributes to the overall HSV-1 anterograde axonal transport, including a major role in virus assembly at the axon terminus, which is not essential in the neuronal cell body. Overall, our data suggest that the process of virus assembly at the growth cones differs from that in the neuronal cell body and that HSV-1 has evolved different mechanisms for virus assembly and exit from different cellular compartments.

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Primary cilia safeguard cortical neurons in neonatal mouse forebrain from environmental stress-induced dendritic degeneration

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.2012482118 ◽

2020 ◽

Vol 118 (1) ◽

pp. e2012482118

Author(s):

Seiji Ishii ◽

Toru Sasaki ◽

Shahid Mohammad ◽

Hye Hwang ◽

Edwin Tomy ◽

...

Keyword(s):

Environmental Stress ◽

Cortical Neurons ◽

Primary Cilia ◽

Wide Spectrum ◽

Neuronal Cell ◽

Cytoskeletal Proteins ◽

Neonatal Mouse ◽

Developing Brain ◽

Environmental Stressors ◽

Neuronal Cell Body

The developing brain is under the risk of exposure to a multitude of environmental stressors. While perinatal exposure to excessive levels of environmental stress is responsible for a wide spectrum of neurological and psychiatric conditions, the developing brain is equipped with intrinsic cell protection, the mechanisms of which remain unknown. Here we show, using neonatal mouse as a model system, that primary cilia, hair-like protrusions from the neuronal cell body, play an essential role in protecting immature neurons from the negative impacts of exposure to environmental stress. More specifically, we found that primary cilia prevent the degeneration of dendritic arbors upon exposure to alcohol and ketamine, two major cell stressors, by activating cilia-localized insulin-like growth factor 1 receptor and downstream Akt signaling. We also found that activation of this pathway inhibits Caspase-3 activation and caspase-mediated cleavage/fragmentation of cytoskeletal proteins in stress-exposed neurons. These results indicate that primary cilia play an integral role in mitigating adverse impacts of environmental stressors such as drugs on perinatal brain development.

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The morphology of the sensory neuron in the cockroach femoral tactile spine

Journal of Neurophysiology ◽

10.1152/jn.1993.69.3.669 ◽

1993 ◽

Vol 69 (3) ◽

pp. 669-673 ◽

Cited By ~ 8

Author(s):

A. S. French ◽

A. R. Klimaszewski ◽

L. L. Stockbridge

Keyword(s):

Action Potential ◽

Sensory Neuron ◽

Cell Body ◽

Computer Software ◽

Neuronal Cell ◽

Neuronal Morphology ◽

Neuronal Cell Body ◽

Intracellular Recordings ◽

Neuronal Nucleus ◽

Spine Structure

1. The cockroach femoral tactile spine contains a single bipolar sensory neuron. The mechanosensitive dendrite in the wall of the spine leads through the spine lumen to a cell body, and then to an axon that proceeds proximally along the femur. The ultrastructure of the sensory ending has been examined before with electron microscopy. However, the morphology of the complete neuron and its relationship to the general spine structure have not been described before. 2. The tactile spine neuron has been extensively used in electrophysiological studies, including intracellular recordings. Action-potential amplitudes and thresholds were variable and inversely related in intracellular recordings, which could be caused by variability in the location of the action-potential initiation region, the position of the recording electrode, or the neuronal morphology. Attempts to observe the complete neuronal morphology by dye injection were hampered by the opaque and autofluorescent cuticle surrounding the neuron. 3. We examined 10 tactile spine neurons, and their surrounding structures, by taking serial 1-micron sections through the base of the spine, normal to its long axis. The sections were examined with light microscopy, digitized by tracing onto a graphics tablet, and then reassembled with the use of computer software. Reconstructions were made of the borders of the spine cuticle, neuron, neuronal nucleus, glial wrappings, and the main trachea in the spine lumen. 4. There was considerable variability in the size and shape of the neuronal cell body, although the sensory dendrite and axon had more consistent morphologies.(ABSTRACT TRUNCATED AT 250 WORDS)

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Role of Exendin-4 in Brain Insulin Resistance, Mitochondrial Function, and Neurite Outgrowth in Neurons under Palmitic Acid-Induced Oxidative Stress

Antioxidants ◽

10.3390/antiox10010078 ◽

2021 ◽

Vol 10 (1) ◽

pp. 78

Author(s):

Danbi Jo ◽

Gwangho Yoon ◽

Juhyun Song

Keyword(s):

Insulin Resistance ◽

Glucose Metabolism ◽

Palmitic Acid ◽

Cortical Neurons ◽

Neuronal Damage ◽

Neuronal Cell ◽

Neuronal Morphology ◽

Therapeutic Drug ◽

Signal Pathways ◽

Impaired Glucose Metabolism

Glucagon like peptide 1 (GLP-1) is an incretin hormone produced by the gut and brain, and is currently being used as a therapeutic drug for type 2 diabetes and obesity, suggesting that it regulates abnormal appetite patterns, and ameliorates impaired glucose metabolism. Many researchers have demonstrated that GLP-1 agonists and GLP-1 receptor agonists exert neuroprotective effects against brain damage. Palmitic acid (PA) is a saturated fatty acid, and increases the risk of neuroinflammation, lipotoxicity, impaired glucose metabolism, and cognitive decline. In this study, we investigated whether or not Exentin-4 (Ex-4; GLP-1 agonist) inhibits higher production of reactive oxygen species (ROS) in an SH-SY5Y neuronal cell line under PA-induced apoptosis conditions. Moreover, pre-treatment with Ex-4 in SH-SY5Y neuronal cells prevents neural apoptosis and mitochondrial dysfunction through several cellular signal pathways. In addition, insulin sensitivity in neurons is improved by Ex-4 treatment under PA-induced insulin resistance. Additionally, our imaging data showed that neuronal morphology is improved by EX-4 treatment, in spite of PA-induced neuronal damage. Furthermore, we identified that Ex-4 inhibits neuronal damage and enhanced neural complexity, such as neurite length, secondary branches, and number of neurites from soma in PA-treated SH-SY5Y. We observed that Ex-4 significantly increases neural complexity, dendritic spine morphogenesis, and development in PA treated primary cortical neurons. Hence, we suggest that GLP-1 administration may be a crucial therapeutic solution for improving neuropathology in the obese brain.

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Epigenomic diversity of cortical projection neurons in the mouse brain

Nature ◽

10.1038/s41586-021-03223-w ◽

2021 ◽

Vol 598 (7879) ◽

pp. 167-173 ◽

Cited By ~ 4

Author(s):

Zhuzhu Zhang ◽

Jingtian Zhou ◽

Pengcheng Tan ◽

Yan Pang ◽

Angeline C. Rivkin ◽

...

Keyword(s):

Single Cell ◽

Cortical Neurons ◽

Neuronal Cell ◽

Cell Types ◽

Molecular Properties ◽

Projection Neurons ◽

Long Distance ◽

Cortical Projection ◽

Retrograde Labelling ◽

Projection Properties

AbstractNeuronal cell types are classically defined by their molecular properties, anatomy and functions. Although recent advances in single-cell genomics have led to high-resolution molecular characterization of cell type diversity in the brain1, neuronal cell types are often studied out of the context of their anatomical properties. To improve our understanding of the relationship between molecular and anatomical features that define cortical neurons, here we combined retrograde labelling with single-nucleus DNA methylation sequencing to link neural epigenomic properties to projections. We examined 11,827 single neocortical neurons from 63 cortico-cortical and cortico-subcortical long-distance projections. Our results showed unique epigenetic signatures of projection neurons that correspond to their laminar and regional location and projection patterns. On the basis of their epigenomes, intra-telencephalic cells that project to different cortical targets could be further distinguished, and some layer 5 neurons that project to extra-telencephalic targets (L5 ET) formed separate clusters that aligned with their axonal projections. Such separation varied between cortical areas, which suggests that there are area-specific differences in L5 ET subtypes, which were further validated by anatomical studies. Notably, a population of cortico-cortical projection neurons clustered with L5 ET rather than intra-telencephalic neurons, which suggests that a population of L5 ET cortical neurons projects to both targets. We verified the existence of these neurons by dual retrograde labelling and anterograde tracing of cortico-cortical projection neurons, which revealed axon terminals in extra-telencephalic targets including the thalamus, superior colliculus and pons. These findings highlight the power of single-cell epigenomic approaches to connect the molecular properties of neurons with their anatomical and projection properties.

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Epigenomic Diversity of Cortical Projection Neurons in the Mouse Brain

10.1101/2020.04.01.019612 ◽

2020 ◽

Cited By ~ 3

Author(s):

Zhuzhu Zhang ◽

Jingtian Zhou ◽

Pengcheng Tan ◽

Yan Pang ◽

Angeline Rivkin ◽

...

Keyword(s):

Single Cell ◽

Cortical Neurons ◽

Neuronal Cell ◽

Cell Types ◽

Molecular Properties ◽

Projection Neurons ◽

Long Distance ◽

Cortical Projection ◽

Axon Terminals ◽

Projection Properties

SummaryNeuronal cell types are classically defined by their molecular properties, anatomy, and functions. While recent advances in single-cell genomics have led to high-resolution molecular characterization of cell type diversity in the brain, neuronal cell types are often studied out of the context of their anatomical properties. To better understand the relationship between molecular and anatomical features defining cortical neurons, we combined retrograde labeling with single-nucleus DNA methylation sequencing to link epigenomic properties of cell types to neuronal projections. We examined 11,827 single neocortical neurons from 63 cortico-cortical (CC) and cortico-subcortical long-distance projections. Our results revealed unique epigenetic signatures of projection neurons that correspond to their laminar and regional location and projection patterns. Based on their epigenomes, intra-telencephalic (IT) cells projecting to different cortical targets could be further distinguished, and some layer 5 neurons projecting to extra-telencephalic targets (L5-ET) formed separate subclusters that aligned with their axonal projections. Such separation varied between cortical areas, suggesting area-specific differences in L5-ET subtypes, which were further validated by anatomical studies. Interestingly, a population of CC projection neurons clustered with L5-ET rather than IT neurons, suggesting a population of L5-ET cortical neurons projecting to both targets (L5-ET+CC). We verified the existence of these neurons by labeling the axon terminals of CC projection neurons and observed clear labeling in ET targets including thalamus, superior colliculus, and pons. These findings highlight the power of single-cell epigenomic approaches to connect the molecular properties of neurons with their anatomical and projection properties.

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