ABSTRACT
Soluble amyloid-beta (Abeta) peptide is likely to play a key role during early stages of Alzheimer's disease (AD) by perturbing synaptic function and cognitive processes. Receptor for advanced glycation end products (RAGE) has been identified as a receptor involved in Abeta-induced neuronal dysfunction. We investigated the role of neuronal RAGE in Abeta-induced synaptic dysfunction in the entorhinal cortex, an area of the brain important in memory processes that is affected early in AD. We found that soluble oligomeric Abeta peptide (Abeta42) blocked long-term potentiation (LTP), but did not affect long-term depression, paired-pulse facilitation, or basal synaptic transmission. In contrast, Abeta did not inhibit LTP in slices from RAGE-null mutant mice or in slices from wild-type mice treated with anti-RAGE IgG. Similarly, transgenic mice expressing a dominant-negative form of RAGE targeted to neurons showed normal LTP in the presence of Abeta, suggesting that neuronal RAGE functions as a signal transducer for Abeta-mediated LTP impairment. To investigate intracellular pathway transducing RAGE activation by Abeta, we used inhibitors of stress activated kinases. We found that inhibiting p38 mitogen-activated protein kinase (p38 MAPK), but not blocking c-Jun N-terminal kinase activation, was capable of maintaining LTP in Abeta-treated slices. Moreover, Abeta-mediated enhancement of p38 MAPK phosphorylation in cortical neurons was reduced by blocking antibodies to RAGE. Together, our results indicate that Abeta impairs LTP in the entorhinal cortex through neuronal RAGE-mediated activation of p38 MAPK.
Subject(s)
Amyloid beta-Peptides/toxicity , Neurons/cytology , Neurons/drug effects , Peptide Fragments/toxicity , Receptors, Immunologic/metabolism , Synapses/physiology , p38 Mitogen-Activated Protein Kinases/physiology , Action Potentials/physiology , Action Potentials/radiation effects , Animals , Animals, Newborn , Antibodies/pharmacology , Cells, Cultured , Dose-Response Relationship, Radiation , Electric Stimulation/methods , Entorhinal Cortex/cytology , Enzyme Activation , Enzyme-Linked Immunosorbent Assay/methods , In Vitro Techniques , Long-Term Potentiation/drug effects , Long-Term Potentiation/radiation effects , Mice , Mice, Inbred C57BL , Mice, Transgenic , Neural Inhibition/drug effects , Neural Inhibition/radiation effects , Receptor for Advanced Glycation End Products , Receptors, Immunologic/genetics , Receptors, Immunologic/immunology , Synapses/drug effectsABSTRACT
During neuronal differentiation, lamellipodia and filopodia explore the environment in search for the correct path to the axon's final destination. Although the motion of lamellipodia and filopodia has been characterized to an extent, little is known about the force they exert. In this study, we used optical tweezers to measure the force exerted by filopodia and lamellipodia with a millisecond temporal resolution. We found that a single filopodium exerts a force not exceeding 3 pN, whereas lamellipodia can exert a force up to 20 pN. Using metabolic inhibitors, we showed that no force is produced in the absence of actin polymerization and that development of forces larger than 3 pN requires microtubule polymerization. These results show that actin polymerization is necessary for force production and demonstrate that not only do neurons process information, but they also act on their environment exerting forces varying from tenths pN to tens of pN.
Subject(s)
Cytoskeleton/metabolism , Pseudopodia/metabolism , Actins/metabolism , Animals , Axons/metabolism , Cell Movement , Cytoskeletal Proteins/metabolism , Extracellular Matrix/metabolism , Growth Cones/metabolism , Models, Biological , Models, Statistical , Neurons/metabolism , Optical Tweezers , RatsABSTRACT
The microtubule binding protein gephyrin plays a prominent role in establishing and maintaining a high concentration of inhibitory glycine receptors juxtaposed to presynaptic releasing sites. Here, we show that endogenous gephyrin undergoes proline-directed phosphorylation, which is followed by the recruitment of the peptidyl-prolyl isomerase Pin1. The interaction between gephyrin and Pin1 is strictly dependent on gephyrin phosphorylation and requires serine-proline consensus sites encompassing the gephyrin proline-rich domain. Upon binding, Pin1 triggers conformational changes in the gephyrin molecule, thus enhancing its ability to bind the beta subunit of GlyRs. Consistently, a downregulation of GlyR clusters was detected in hippocampal neurons derived from Pin1 knockout mice, which was paralleled by a reduction in the amplitude of glycine-evoked currents. Our results suggest that phosphorylation-dependent prolyl isomerisation of gephyrin represents a mechanism for regulating GlyRs function.
Subject(s)
Carrier Proteins/metabolism , Membrane Proteins/metabolism , Peptidylprolyl Isomerase/metabolism , Receptors, Glycine/metabolism , Animals , Brain/metabolism , Carrier Proteins/chemistry , Cytoplasmic Structures/metabolism , Epitopes , Evoked Potentials , Hippocampus/cytology , Hippocampus/enzymology , Humans , Membrane Proteins/chemistry , Mice , Mice, Knockout , NIMA-Interacting Peptidylprolyl Isomerase , Neurons/cytology , Neurons/enzymology , Peptidylprolyl Isomerase/deficiency , Phosphorylation , Proline/metabolism , Protein Binding , Protein Structure, Tertiary , Protein Subunits/metabolism , Recombinant Proteins/metabolism , Serine/metabolismABSTRACT
We demonstrate the possibility of using carbon nanotubes (CNTs) as potential devices able to improve neural signal transfer while supporting dendrite elongation and cell adhesion. The results strongly suggest that the growth of neuronal circuits on a CNT grid is accompanied by a significant increase in network activity. The increase in the efficacy of neural signal transmission may be related to the specific properties of CNT materials, such as the high electrical conductivity.
