scholarly journals Modulation of receptor cycling by neuron-enriched endosomal protein of 21 kD

2002 ◽  
Vol 157 (7) ◽  
pp. 1197-1209 ◽  
Author(s):  
Pascal Steiner ◽  
J.-C. Floyd Sarria ◽  
Liliane Glauser ◽  
Sarah Magnin ◽  
Stefan Catsicas ◽  
...  
Keyword(s):  
Hippocampal Neurons ◽  
Neuronal Membrane ◽  
Primary Neurons ◽  
Receptor Recycling ◽  
Down Regulation ◽  
Developmentally Regulated ◽  
Neuronal Protein ◽  
Specific Proteins ◽  
Endosomal Pathway ◽  
Stimulation Of

Although correct cycling of neuronal membrane proteins is essential for neurite outgrowth and synaptic plasticity, neuron-specific proteins of the implicated endosomes have not been characterized. Here we show that a previously cloned, developmentally regulated, neuronal protein of unknown function binds to syntaxin 13. We propose to name this protein neuron-enriched endosomal protein of 21 kD (NEEP21), because it is colocalized with transferrin receptors, internalized transferrin (Tf), and Rab4. In PC12 cells, NEEP21 overexpression accelerates Tf internalization and recycling, whereas its down-regulation strongly delays Tf recycling. In primary neurons, NEEP21 is localized to the somatodendritic compartment, and, upon N-methyl-d-aspartate (NMDA) stimulation, the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor subunit GluR2 is internalized into NEEP21-positive endosomes. NEEP21 down-regulation retards recycling of GluR1 to the cell surface after NMDA stimulation of hippocampal neurons. In summary, NEEP21 is a neuronal protein that is localized to the early endosomal pathway and is necessary for correct receptor recycling in neurons.

A developmentally regulated switch in neuronal responsiveness to NCAM and N-cadherin in the rat hippocampus

Development ◽  
1992 ◽  
Vol 115 (3) ◽  
pp. 885-892 ◽  
Author(s):  
P. Doherty ◽  
S.D. Skaper ◽  
S.E. Moore ◽  
A. Leon ◽  
F.S. Walsh
Keyword(s):  
Neurite Outgrowth ◽  
Cell Adhesion Molecules ◽  
Hippocampal Neurons ◽  
Direct Evidence ◽  
Polysialic Acid ◽  
Primary Neurons ◽  
Calcium Channel Antagonists ◽  
3T3 Cells ◽  
Developmentally Regulated ◽  
3T3 Fibroblasts

Monolayers of control 3T3 fibroblasts and 3T3 cells expressing transfected NCAM or N-cadherin have been used as a culture substratum for rat hippocampal neurons. Both NCAM and N-cadherin are expressed in the hippocampus through embryonic day 17 (E17) to postnatal day 4 (PND4); however, whereas E17 neurons responded to transfected NCAM by extending considerably longer neurites, PND4 neurons responded very poorly. The converse was true for responsiveness to N-cadherin. These data demonstrate a switch in neuronal responsiveness to NCAM and N-cadherin in the developing hippocampus. NCAM-dependent neurite outgrowth from E17 neurons was largely dependent on the presence of alpha 2–8-linked polysialic acid (PSA) on neuronal NCAM. NCAM-dependent neurite outgrowth could be fully inhibited by pertussis toxin or a combination of L- and N-type calcium channel antagonists thus providing direct evidence concerning the nature of the second messenger pathway activated in primary neurons by cell adhesion molecules (CAMs).

scholarly journals A Super-Resolved View of the Alzheimer’s Disease-Related Amyloidogenic Pathway in Hippocampal Neurons

2021 ◽  
pp. 1-20
Author(s):  
Yang Yu ◽  
Yang Gao ◽  
Bengt Winblad ◽  
Lars Tjernberg ◽  
Sophia Schedin Weiss
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Hippocampal Neurons ◽  
Three Dimensional ◽  
Amyloid Β ◽  
Stimulated Emission ◽  
Amyloid Β Peptide ◽  
Primary Neurons ◽  
Amyloid Β Protein ◽  
Early Endosomes

Background: Processing of the amyloid-β protein precursor (AβPP) is neurophysiologically important due to the resulting fragments that regulate synapse biology, as well as potentially harmful due to generation of the 42 amino acid long amyloid β-peptide (Aβ 42), which is a key player in Alzheimer’s disease. Objective: Our aim was to clarify the subcellular locations of the amyloidogenic AβPP processing in primary neurons, including the intracellular pools of the immediate substrate, AβPP C-terminal fragment (APP-CTF) and the product (Aβ 42). To overcome the difficulties of resolving these compartments due to their small size, we used super-resolution microscopy. Methods: Mouse primary hippocampal neurons were immunolabelled and imaged by stimulated emission depletion (STED) microscopy, including three-dimensional, three-channel imaging and image analyses. Results: The first (β-secretase) and second (γ-secretase) cleavages of AβPP were localized to functionally and distally distinct compartments. The β-secretase cleavage was observed in early endosomes, where we were able to show that the liberated N- and C-terminal fragments were sorted into distinct vesicles budding from the early endosomes in soma. Lack of colocalization of Aβ 42 and APP-CTF in soma suggested that γ-secretase cleavage occurs in neurites. Indeed, APP-CTF was, in line with Aβ 42 in our previous study, enriched in the presynapse but absent from the postsynapse. In contrast, full-length AβPP was not detected in either the pre- or the postsynaptic side of the synapse. Furthermore, we observed that endogenously produced and endocytosed Aβ 42 were localized in different compartments. Conclusion: These findings provide critical super-resolved insight into amyloidogenic AβPP processing in primary neurons.

scholarly journals Induction of Axonal Outgrowth in Mouse Hippocampal Neurons via Bacterial Magnetosomes

2021 ◽  
Vol 22 (8) ◽  
pp. 4126
Author(s):  
Sara De Vincentiis ◽  
Alessandro Falconieri ◽  
Frank Mickoleit ◽  
Valentina Cappello ◽  
Dirk Schüler ◽  
...  
Keyword(s):  
Magnetic Properties ◽  
Iron Oxide ◽  
Hippocampal Neurons ◽  
Magnetotactic Bacteria ◽  
Oxide Nanoparticles ◽  
Axonal Outgrowth ◽  
Mechanical Forces ◽  
Internalization Process ◽  
Oxide Crystals ◽  
Stimulation Of

Magnetosomes are membrane-enclosed iron oxide crystals biosynthesized by magnetotactic bacteria. As the biomineralization of bacterial magnetosomes can be genetically controlled, they have become promising nanomaterials for bionanotechnological applications. In the present paper, we explore a novel application of magnetosomes as nanotool for manipulating axonal outgrowth via stretch-growth (SG). SG refers to the process of stimulation of axonal outgrowth through the application of mechanical forces. Thanks to their superior magnetic properties, magnetosomes have been used to magnetize mouse hippocampal neurons in order to stretch axons under the application of magnetic fields. We found that magnetosomes are avidly internalized by cells. They adhere to the cell membrane, are quickly internalized, and slowly degrade after a few days from the internalization process. Our data show that bacterial magnetosomes are more efficient than synthetic iron oxide nanoparticles in stimulating axonal outgrowth via SG.

Differential Oxidative Modulation of Voltage-Dependent K+ Currents in Rat Hippocampal Neurons

2002 ◽  
Vol 87 (6) ◽  
pp. 2990-2995 ◽  
Author(s):  
Wolfgang Müller ◽  
Katrin Bittner
Keyword(s):  
Hydrogen Peroxide ◽  
Hippocampal Neurons ◽  
Immune Defense ◽  
Delayed Rectifier ◽  
Delayed Rectifier Current ◽  
Cell Recording ◽  
Voltage Dependent ◽  
Steady State Inactivation ◽  
K Currents ◽  
Stimulation Of

Oxidative stress is enhanced by [Ca2+]i-dependent stimulation of phospholipases and mitochondria and has been implicated in immune defense, ischemia, and excitotoxicity. Using whole cell recording from hippocampal neurons, we show that arachidonic acid (AA) and hydrogen peroxide (H2O2) both reduce the transient K+ current I A by −54 and −68%, respectively, and shift steady-state inactivation by −10 and −15 mV, respectively. While AA was effective at an extracellular concentration of 1 μM and an intracellular concentration of 1 pM, extracellular H2O2 was equally effective only at a concentration >800 μM (0.0027%). In contrast to AA, H2O2 decreased the slope of activation and increased the slope of inactivation of I A and reduced the sustained delayed rectifier current I K(V) by 22% and shifted its activation by −9 mV. Intracellular application of the antioxidant glutathione (GSH, 2–5 mM) blocked all effects of AA and the reduction of I A by H2O2. In contrast, intracellular GSH enhanced reduction of I K(V) by H2O2. Decrease of the slope of activation and increase of the slope of inactivation of I A by hydrogen peroxide was blocked and reversed to a decrease, respectively, by intracellular application of GSH. Intracellular GSH did not prevent H2O2 to shift inactivation and activation of I A and activation of I K(V) to more negative potentials. We conclude, that AA and H2O2modulate voltage-activated K currents differentially by oxidation of GSH accessible intracellular and GSH inaccessible extracellular K+-channel domains, thereby presumably affecting neuronal information processing and oxidative damage.

Desensitization and resensitization of lutropin receptors expressed in transfected Y-1 adrenal cells

1999 ◽  
Vol 163 (2) ◽  
pp. 289-297 ◽  
Author(s):  
GA Ulaner ◽  
J Chuang ◽  
W Lin ◽  
D Woodbury ◽  
RV Myers ◽  
...  
Keyword(s):  
Receptor Occupancy ◽  
Untranslated Regions ◽  
Protein Synthesis Inhibitor ◽  
Receptor Recycling ◽  
Synthesis Inhibitor ◽  
Early Step ◽  
Hormonal Stimulation ◽  
Adrenal Cells ◽  
Metallothionein Promoter ◽  
Stimulation Of

Stimulation of gonadal cells by lutropins such as human chorionic gonadotropin (hCG) is often transient and followed by down-regulation and/or desensitization of lutropin receptors (LHR). Here we describe desensitization/resensitization of LHR in Y-1 adrenal cell lines (termed Y-1L) expressing a rat cDNA lacking most 5' and 3' LHR untranslated regions under the control of a metallothionein promoter. Using a simple morphological assay in which stimulated cells are round and unstimulated cells are flat, we identified clones that rounded and remained round and others that became insensitive to lutropin stimulation and reverted to their flat appearance within 2-4 h. Flattened cells were insensitive to further hormonal stimulation but rounded after treatments with cholera toxin, forskolin, or cyclic AMP, showing that loss of responsiveness was associated with an early step in signal transduction, not loss of rounding potential. Removing the lutropin stimulus for at least 90-120 min reversed hormone insensitivity, even in the presence of the protein synthesis inhibitor puromycin. The number of surface bound receptors did not change during a cycle of rounding/flattening and hCG bound to rounded or flattened cells was replaced equally by radioiodinated hCG during incubations at 4 degrees C. Thus, desensitization/resensitization of LHR in Y-1L cells occurred in the absence of new receptor synthesis, receptor degradation, or receptor recycling. These observations suggest that LHR desensitization/resensitization in Y-1L cells was closely coupled to receptor occupancy and that this cell line may be useful for identifying factors that modulate the activities of occupied receptors.

Dynamics of Excitatory Transmitter Release: Analysis of Synaptic Responses in CA3 Hippocampal Neurons After Repetitive Stimulation of Afferent Fibers

1998 ◽  
Vol 79 (4) ◽  
pp. 1977-1988 ◽  
Author(s):  
Marco Canepari ◽  
Enrico Cherubini
Keyword(s):  
Transmitter Release ◽  
Hippocampal Neurons ◽  
Repetitive Stimulation ◽  
Patch Clamp Technique ◽  
Afferent Fibers ◽  
Presynaptic Mechanisms ◽  
Release Analysis ◽  
Excitatory Transmitter ◽  
Stimulation Of ◽  
Synaptic Responses

Canepari, Marco and Enrico Cherubini. Dynamics of excitatory transmitter release: analysis of synaptic responses in CA3 hippocampal neurons after repetitive stimulation of afferent fibers. J. Neurophysiol. 79: 1977–1988, 1998. The patch-clamp technique (whole cell configuration) was used to record excitatory postsynaptic currents (EPSCs) evoked by repetitive stimulation (4 pulses at 50-ms intervals) of afferent fibers in the stratum lucidum-radiatum. Different synaptic behaviors (EPSC patterns) were classified in terms of facilitation or depression of the mean amplitude of the second, third, and fourth EPSC with respect to the previous one. A large variety of EPSC patterns was observed by stimulating different afferent fibers. Experiments with the mGluR2/mGluR3 agonist 2-(2,3-dicarboxycyclopropyl)glycine (DCG-IV) (1 μM), a compound that reduces release at mossy but not at associative commissural fibers and therefore allows to identify the origin of synaptic responses, showed that particular EPSC patterns could not be associated to the activation of a specific type of synaptic input. To investigate the role of the probability of release in the dynamics of synaptic activity, the extracellular calcium concentration was varied from 0.8 to 4 mM in several experiments. EPSC patterns dominated by depression, characteristics of high release probability conditions, could be observed in the majority of the cases in the presence of higher calcium concentrations. A quantitative model for dynamics of transmitter release has been developed. Experimental results were compared with data computed with the model taking into account the probability of release and the time course of reavailability. This work indicates that short-term changes of presynaptic conditions occurring during a train of action potentials can account for the high variability of EPSC responses. The model that is proposed also suggests a general method of experimental data analysis to investigate the possible presynaptic mechanisms underlying long-lasting changes in synaptic efficacy.

Stimulation of actin synthesis by cytochalasin D is specific for the isoactins normally expressed in muscle or non-muscle cells

1986 ◽  
Vol 84 (1) ◽  
pp. 253-262
Author(s):  
J. Tannenbaum ◽  
A.F. Miranda
Keyword(s):  
Muscle Cell ◽  
Cell Cultures ◽  
Muscle Cells ◽  
Cytochalasin D ◽  
Human Muscle ◽  
The State ◽  
Cytoskeletal Proteins ◽  
Developmentally Regulated ◽  
Alpha Actin ◽  
Stimulation Of

Treatment of human muscle myotube cultures with 2 microM-cytochalasin D (CD) for 6 h stimulated synthesis of both the (muscle-specific) alpha-actin and the (non-muscle) beta and gamma-actins usually expressed by these cells. In non-muscle (HEp-2) cell cultures, CD enhanced synthesis of beta and gamma-actin, but did not induce synthesis of alpha-actin, which is not normally present in these cells. Thus, synthesis of both muscle and non-muscle actins can be increased by CD, but enhancement of actin synthesis results from increases in the isoactins usually present, rather than induction of new isotypes. Comparison of CD-treated (fused) myotube cultures with (unfused) myoblast cultures indicated that beta and gamma-actin synthesis was similarly enhanced in both cultures, but that alpha-actin synthesis was stimulated to a greater extent in the myoblast cultures. Desmin synthesis was also stimulated in the myoblasts but not the myotubes, suggesting that the effect of CD on synthesis of these developmentally regulated cytoskeletal proteins (alpha-actin, desmin) might be modulated by fusion or the state of differentiation of the muscle cell.

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