scholarly journals The schizophrenia susceptibility factor dysbindin and its associated complex sort cargoes from cell bodies to the synapse

2011 ◽  
Vol 22 (24) ◽  
pp. 4854-4867 ◽  
Author(s):  
Jennifer Larimore ◽  
Karine Tornieri ◽  
Pearl V. Ryder ◽  
Avanti Gokhale ◽  
Stephanie A. Zlatic ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Cortical Neurons ◽  
Neuronal Cells ◽  
Neuronal Cell ◽  
Adaptor Protein ◽  
Nerve Terminals ◽  
Endosome Trafficking ◽  
Neuronal Cell Bodies ◽  
Cultured Cortical Neurons ◽  
Lysosome Related Organelles

Dysbindin assembles into the biogenesis of lysosome-related organelles complex 1 (BLOC-1), which interacts with the adaptor protein complex 3 (AP-3), mediating a common endosome-trafficking route. Deficiencies in AP-3 and BLOC-1 affect synaptic vesicle composition. However, whether AP-3-BLOC-1–dependent sorting events that control synapse membrane protein content take place in cell bodies upstream of nerve terminals remains unknown. We tested this hypothesis by analyzing the targeting of phosphatidylinositol-4-kinase type II α (PI4KIIα), a membrane protein present in presynaptic and postsynaptic compartments. PI4KIIα copurified with BLOC-1 and AP-3 in neuronal cells. These interactions translated into a decreased PI4KIIα content in the dentate gyrus of dysbindin-null BLOC-1 deficiency and AP-3–null mice. Reduction of PI4KIIα in the dentate reflects a failure to traffic from the cell body. PI4KIIα was targeted to processes in wild-type primary cultured cortical neurons and PC12 cells but failed to reach neurites in cells lacking either AP-3 or BLOC-1. Similarly, disruption of an AP-3–sorting motif in PI4KIIα impaired its sorting into processes of PC12 and primary cultured cortical neuronal cells. Our findings indicate a novel vesicle transport mechanism requiring BLOC-1 and AP-3 complexes for cargo sorting from neuronal cell bodies to neurites and nerve terminals.

scholarly journals Extracellular vesicles from amyloid-β exposed cell cultures induce severe dysfunction in cortical neurons

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Chiara Beretta ◽  
Elisabeth Nikitidou ◽  
Linn Streubel-Gallasch ◽  
Martin Ingelsson ◽  
Dag Sehlin ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Cortical Neurons ◽  
Amyloid Β ◽  
Neuronal Cell ◽  
Time Lapse ◽  
Cellular Mechanisms ◽  
Axonal Swelling ◽  
Tem Analysis ◽  
Neuronal Cell Bodies ◽  
Time Lapse Imaging

AbstractAlzheimer’s disease (AD) is characterized by a substantial loss of neurons and synapses throughout the brain. The exact mechanism behind the neurodegeneration is still unclear, but recent data suggests that spreading of amyloid-β (Aβ) pathology via extracellular vesicles (EVs) may contribute to disease progression. We have previously shown that an incomplete degradation of Aβ42 protofibrils by astrocytes results in the release of EVs containing neurotoxic Aβ. Here, we describe the cellular mechanisms behind EV-associated neurotoxicity in detail. EVs were isolated from untreated and Aβ42 protofibril exposed neuroglial co-cultures, consisting mainly of astrocytes. The EVs were added to cortical neurons for 2 or 4 days and the neurodegenerative processes were followed with immunocytochemistry, time-lapse imaging and transmission electron microscopy (TEM). Addition of EVs from Aβ42 protofibril exposed co-cultures resulted in synaptic loss, severe mitochondrial impairment and apoptosis. TEM analysis demonstrated that the EVs induced axonal swelling and vacuolization of the neuronal cell bodies. Interestingly, EV exposed neurons also displayed pathological lamellar bodies of cholesterol deposits in lysosomal compartments. Taken together, our data show that the secretion of EVs from Aβ exposed cells induces neuronal dysfunction in several ways, indicating a central role for EVs in the progression of Aβ-induced pathology.

Young’s Modulus of Cortical and P19 Derived Neurons Measured by Atomic Force Microscopy

2012 ◽  
Vol 1420 ◽  
Author(s):  
Elise Spedden ◽  
James D. White ◽  
David Kaplan ◽  
Cristian Staii
Keyword(s):  
Young’S Modulus ◽  
Cortical Neurons ◽  
Young's Modulus ◽  
Neuronal Cell ◽  
Short Term ◽  
Force Microscopy ◽  
Protein Substrates ◽  
Atomic Force ◽  
Neuronal Cell Bodies ◽  
Bulk Tissue

ABSTRACTIn this paper we use the Atomic Force Microscope to measure the Young’s modulus for two types of neuronal cell bodies: cortical neurons obtained from rat embryos and neurons derived from P19 mouse embryonic carcinoma stem cells. The neurons are plated on different substrates coated with two types of protein growth factors, poly-D-lysine and laminin. We report on the Young’s modulus of each type of neuron as well as the variation of modulus between cells plated on different protein substrates. We compare these results to various individual cell and bulk tissue measurements reported in literature. We additionally report on an observed change in the Young’s modulus of cortical neurons when subjected to a short-term reduction in ambient temperature.

scholarly journals Ischemic Cerebral Endothelial Cell–Derived Exosomes Promote Axonal Growth

Stroke ◽  
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.

scholarly journals Protective effects of adenosine on apoptotic neuronal cell death in cultured cortical neurons

2000 ◽  
Vol 82 ◽  
pp. 182
Author(s):  
Yutaka Tamura ◽  
Taizo Fukui ◽  
Megumi Kajikawa ◽  
Mikiko Omoto ◽  
Hirohito Shiomi
Keyword(s):  
Cell Death ◽  
Cortical Neurons ◽  
Neuronal Cell Death ◽  
Neuronal Cell ◽  
Protective Effects ◽  
Cultured Cortical Neurons

scholarly journals Mapping the Proteome of the Synaptic Cleft through Proximity Labeling Reveals New Cleft Proteins

Proteomes ◽  
2018 ◽  
Vol 6 (4) ◽  
pp. 48 ◽  
Author(s):  
Tony Cijsouw ◽  
Austin Ramsey ◽  
TuKiet Lam ◽  
Beatrice Carbone ◽  
Thomas Blanpied ◽  
...  
Keyword(s):  
Cortical Neurons ◽  
Functional Organization ◽  
Neuronal Cell ◽  
Super Resolution ◽  
Synaptic Cleft ◽  
Surface Proteins ◽  
Mammalian Brain ◽  
Label Free ◽  
Edge Zone ◽  
Cultured Cortical Neurons

Synapses are specialized neuronal cell-cell contacts that underlie network communication in the mammalian brain. Across neuronal populations and circuits, a diverse set of synapses is utilized, and they differ in their molecular composition to enable heterogenous connectivity patterns and functions. In addition to pre- and post-synaptic specializations, the synaptic cleft is now understood to be an integral compartment of synapses that contributes to their structural and functional organization. Aiming to map the cleft proteome, this study applied a peroxidase-mediated proximity labeling approach and used the excitatory synaptic cell adhesion protein SynCAM 1 fused to horseradish peroxidase (HRP) as a reporter in cultured cortical neurons. This reporter marked excitatory synapses as measured by confocal microcopy and was targeted to the edge zone of the synaptic cleft as determined using 3D dSTORM super-resolution imaging. Proximity labeling with a membrane-impermeant biotin-phenol compound restricted labeling to the cell surface, and Label-Free Quantitation (LFQ) mass spectrometry combined with ratiometric HRP tagging of membrane vs. synaptic surface proteins was used to identify the proteomic content of excitatory clefts. Novel cleft candidates were identified, and Receptor-type tyrosine-protein phosphatase zeta was selected and successfully validated. This study supports the robust applicability of peroxidase-mediated proximity labeling for synaptic cleft proteomics and its potential for understanding synapse heterogeneity in health and changes in diseases such as psychiatric disorders and addiction.

Neuroprotection by MAPK/ERK kinase inhibition with U0126 against oxidative stress in a mouse neuronal cell line and rat primary cultured cortical neurons

2000 ◽  
Vol 288 (2) ◽  
pp. 163-166 ◽  
Author(s):  
Takumi Satoh ◽  
Daisaku Nakatsuka ◽  
Yasuyoshi Watanabe ◽  
Izumi Nagata ◽  
Haruhiko Kikuchi ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Line ◽  
Cortical Neurons ◽  
Neuronal Cell ◽  
Kinase Inhibition ◽  
Neuronal Cell Line ◽  
Cultured Cortical Neurons ◽  
Erk Kinase

scholarly journals BLOC-1 Interacts with BLOC-2 and the AP-3 Complex to Facilitate Protein Trafficking on Endosomes

2006 ◽  
Vol 17 (9) ◽  
pp. 4027-4038 ◽  
Author(s):  
Santiago M. Di Pietro ◽  
Juan M. Falcón-Pérez ◽  
Danièle Tenza ◽  
Subba R.G. Setty ◽  
Michael S. Marks ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Protein Trafficking ◽  
Adaptor Protein ◽  
Early Endosome ◽  
Related Protein ◽  
Genes Encoding ◽  
Cellular Machinery ◽  
Hermansky Pudlak Syndrome ◽  
Lysosome Related Organelles ◽  
Tyrosinase Related Protein

The adaptor protein (AP)-3 complex is a component of the cellular machinery that controls protein sorting from endosomes to lysosomes and specialized related organelles such as melanosomes. Mutations in an AP-3 subunit underlie a form of Hermansky-Pudlak syndrome (HPS), a disorder characterized by abnormalities in lysosome-related organelles. HPS in humans can also be caused by mutations in genes encoding subunits of three complexes of unclear function, named biogenesis of lysosome-related organelles complex (BLOC)-1, -2, and -3. Here, we report that BLOC-1 interacts physically and functionally with AP-3 to facilitate the trafficking of a known AP-3 cargo, CD63, and of tyrosinase-related protein 1 (Tyrp1), a melanosomal membrane protein previously thought to traffic only independently of AP-3. BLOC-1 also interacts with BLOC-2 to facilitate Tyrp1 trafficking by a mechanism apparently independent of AP-3 function. Both BLOC-1 and -2 localize mainly to early endosome-associated tubules as determined by immunoelectron microscopy. These findings support the idea that BLOC-1 and -2 represent hitherto unknown components of the endosomal protein trafficking machinery.

Localization of Nerve Cells in the Developing Rat Tooth

1997 ◽  
Vol 76 (7) ◽  
pp. 1350-1356 ◽  
Author(s):  
K. Luukko ◽  
K. Sainio ◽  
H. Sariola ◽  
M. Saarma ◽  
I. Thesleff
Keyword(s):  
Neuronal Cells ◽  
Neuronal Cell ◽  
Tooth Germ ◽  
Nerve Fibers ◽  
Tooth Formation ◽  
Neuronal Cell Bodies ◽  
Dental Epithelium ◽  
Tooth Germs ◽  
Developing Rat

Earlier studies have shown that mammalian tooth formation can take place in the absence of peripheral nerve fibers. This has been taken to indicate that neurons are not needed for mammalian tooth development. However, our recent localization of peripherin, which is a neuronal cell marker, has suggested that neuronal cell bodies may be associated with developing teeth. In this study, we have analyzed in vivo and in vitro the presence of neuronal cells in developing rat tooth germs. When E14 and E16 rat first molars (thickening of presumptive dental epithelium and bud-stage tooth germ, respectively) were cultured in vitro, peripheral trigeminal axons degenerated. However, with antibodies against peripherin and L1 neural cell adhesion protein, we detected neuronal cell bodies and their axons in the explants. Next, the expression of neurofilament light-chain (NF-L) mRNAs was studied by in situ hybridization of embryonic E12 first branchial arches and tooth germs from initiation to completion of crown morphogenesis (E13, five-day post-natal teeth). NF-L transcripts were first seen at the bud stage (E15) next to the dental epithelium at the buccal side of the tooth germ. At the cap stage (E18), NF-L mRNAs were located under the oral epithelium at some distance from dental epithelium. These expression patterns correlate to the previous localization of peripherin-positive cells and suggest that NF-L expression also revealed neuronal cells. Taken together, these results demonstrate that, in addition to projections of peripheral neurons, neuronal cells are associated with the developing teeth. Hence, it is possible that neuronal cells may participate in the regulation of mammalian tooth formation.

scholarly journals The Neuron-Specific Rai (ShcC) Adaptor Protein Inhibits Apoptosis by Coupling Ret to the Phosphatidylinositol 3-Kinase/Akt Signaling Pathway

2002 ◽  
Vol 22 (20) ◽  
pp. 7351-7363 ◽  
Author(s):  
Giuliana Pelicci ◽  
Flavia Troglio ◽  
Alessandra Bodini ◽  
Rosa Marina Melillo ◽  
Valentina Pettirossi ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Sympathetic Neurons ◽  
Neuronal Cells ◽  
Neuronal Cell ◽  
Adaptor Protein ◽  
Mitogen Activated Protein Kinase ◽  
Modular Organization ◽  
Phosphatidylinositol 3 Kinase ◽  
Phosphatidylinositol 3

ABSTRACT Rai is a recently identified member of the family of Shc-like proteins, which are cytoplasmic signal transducers characterized by the unique PTB-CH1-SH2 modular organization. Rai expression is restricted to neuronal cells and regulates in vivo the number of postmitotic sympathetic neurons. We report here that Rai is not a common substrate of receptor tyrosine kinases under physiological conditions and that among the analyzed receptors (Ret, epidermal growth factor receptor, and TrkA) it is activated specifically by Ret. Overexpression of Rai in neuronal cell lines promoted survival by reducing apoptosis both under conditions of limited availability of the Ret ligand glial cell line-derived neurotrophic factor (GDNF) and in the absence of Ret activation. Overexpressed Rai resulted in the potentiation of the Ret-dependent activation of phosphatidylinositol 3-kinase (PI3K) and Akt. Notably, increased Akt phosphorylation and PI3K activity were also found under basal conditions, e.g., in serum-starved neuronal cells. Phosphorylated and hypophosphorylated Rai proteins form a constitutive complex with the p85 subunit of PI3K: upon Ret triggering, the Rai-PI3K complex is recruited to the tyrosine-phosphorylated Ret receptor through the binding of the Rai PTB domain to tyrosine 1062 of Ret. In neurons treated with low concentrations of GDNF, the prosurvival effect of Rai depends on Rai phosphorylation and Ret activation. In the absence of Ret activation, the prosurvival effect of Rai is, instead, phosphorylation independent. Finally, we showed that overexpression of Rai, at variance with Shc, had no effects on the early peak of mitogen-activated protein kinase (MAPK) activation, whereas it increased its activation at later time points. Phosphorylated Rai, however, was not found in complexes with Grb2. We propose that Rai potentiates the MAPK and PI3K signaling pathways and regulates Ret-dependent and -independent survival signals.

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