Endocytosis: an assembly protein for clathrin cages.

The protein AP180 is known to have clathrin-assembly activity in vitro. AP180 has now been found to be crucial for synaptic vesicle endocytosis and the maintenance of a uniform-size vesicle population in vivo. These results significantly advance our understanding of clathrin-mediated endocytosis in the synapse and elsewhere.     

AP180 Couples Protein Retrieval to Clathrin‐Mediated Endocytosis of Synaptic Vesicles

How clathrin‐mediated endocytosis (CME) retrieves vesicle proteins into newly formed synaptic vesicles (SVs) remains a major puzzle. Besides its roles in stimulating clathrin‐coated vesicle formation and regulating SV size, the clathrin assembly protein AP180 has been identified as a key player in retrieving SV proteins. The mechanisms by which AP180 recruits SV proteins are not fully understood. Here, we show that following acute inactivation of AP180 in Drosophila, SV recycling is severely impaired at the larval neuromuscular synapse based on analyses of FM 1‐43 uptake and synaptic ultrastructure. More dramatically, AP180 activity is important to maintain the integrity of SV protein complexes at the plasma membrane during endocytosis. These observations suggest that AP180 normally clusters SV proteins together during recycling. Consistent with this notion, SV protein composition and distribution are altered in AP180 mutant flies. Finally, AP180 co‐immunoprecipitates with SV proteins, including the vesicular glutamate transporter and neuronal synaptobrevin. These results reveal a new mode by which AP180 couples protein retrieval to CME of SVs. AP180 is also genetically linked to Alzheimer's disease. Hence, the findings of this study may provide new mechanistic insight into the role of AP180 dysfunction in Alzheimer's disease.      

Serpins are apically secreted from MDCK cells independently of their raft association

The role of phospholipase Cgamma1 (PLCgamma1) in signal transduction was investigated by characterizing its SH domain-binding proteins that may represent components of a novel signaling pathway. A 180-kDa protein that binds to the SH2 domain of PLCgamma1 was purified from rat brain. The amino acid sequence of peptide derived from the purified protein is now identified as AP180, a clathrin assembly protein that has been implicated in clathrin-mediated synaptic vesicle recycling in synapses. In this report, we demonstrate the stable association of PLCgamma1 with AP180 in a clathrin-coated vesicle complex, which not only binds to the carboxyl-terminal SH2 domain of PLCgamma1, but also inhibits its enzymatic activity in a dose-dependent manner.     

Clathrin assembly protein AP180: primary structure, domain organization and identification of a clathrin binding site.

Binding of AP180 to clathrin triskelia induces their assembly into 60-70 nm coats. The largest rat brain cDNA clone isolated predicts a molecular weight of 91,430 for AP180. Two cDNA clones have an additional small 57 bp insert. The deduced molecular weight agrees with gel filtration results provided the more chaotropic denaturant 6 M guanidinium thiocyanate is substituted for the weaker guanidinium chloride. The sequence and the proteolytic cleavage pattern suggest a three domain structure. The N-terminal 300 residues (pI 8.7) harbour a clathrin binding site. An acidic middle domain (pI 3.6, 450 residues), interrupted by an uncharged alanine rich segment of 59 residues, appears to be responsible for the anomalous physical properties of AP180. The C-terminal domain (166 residues) has a pI of 10.4. AP180 mRNA is restricted to neuronal sources. AP180 shows no significant homology to known clathrin binding proteins, but is nearly identical to a mouse phosphoprotein (F1-20). This protein, localized to synaptic termini, has so far been of unknown function.     

Miniature neurotransmission stabilizes synaptic function via tonic suppression of local dendritic protein synthesis

Activity deprivation in neurons induces a slow compensatory scaling up of synaptic strength, reflecting a homeostatic mechanism for stabilizing neuronal activity. Prior studies have focused on the loss of action potential (AP) driven neurotransmission in synaptic homeostasis. Here, we show that the miniature synaptic transmission that persists during AP blockade profoundly shapes the time course and mechanism of homeostatic scaling. A brief blockade of NMDA receptor (NMDAR) mediated miniature synaptic events ("minis") rapidly scales up synaptic strength, over an order of magnitude faster than with AP blockade alone. The rapid scaling induced by NMDAR mini blockade is mediated by increased synaptic expression of surface GluR1 and the transient incorporation of Ca2+-permeable AMPA receptors at synapses; both of these changes are implemented locally within dendrites and require dendritic protein synthesis. These results indicate that NMDAR signaling during miniature synaptic transmission serves to stabilize synaptic function through active suppression of dendritic protein synthesis.     

Activity-dependent wiring of the developing hippocampal neuronal circuit

In the developing hippocampus, functional excitatory synaptic connections seem to be recruited from a preformed, initially silent synaptic network. This functional synapse induction requires presynaptic action potentials paired with postsynaptic depolarization, thus obeying Hebb's rule of association. During early postnatal development the hippocampus exhibits an endogenous form of patterned neuronal activity that is driven by GABAergic depolarization. We propose that this recurrent activity promotes the input-specific induction of functional synapses in the CA1 region. Thus, activity-dependent synaptic reorganization in the developing hippocampus appears to be dominated by an active recruitment of new synapses rather than an active elimination of redundant connections.     

Computational simulation: astrocyte-induced depolarization of neighboring neurons mediates synchronous UP states in a neural network

Although recent reports have suggested that synchronous neuronal UP states are mediated by astrocytic activity, the mechanism responsible for this remains unknown. Astrocytic glutamate release synchronously depolarizes adjacent neurons, while synaptic transmissions are blocked. The purpose of this study was to confirm that astrocytic depolarization, propagated through synaptic connections, can lead to synchronous neuronal UP states. We applied astrocytic currents to local neurons in a neural network consisting of model cortical neurons. Our results show that astrocytic depolarization may generate synchronous UP states for hundreds of milliseconds in neurons even if they do not directly receive glutamate release from the activated astrocyte.     

Membrane Potential Oscillations Produced by Excitatory Amino Acids in Isolated Spinal Neurons of the Lamprey Lampetra fluviatilis

Membrane potential (MP) oscillations produced by excitatory amino acids (EAA) have been studied in branching neurons isolated by an enzymatic-mechanical method from the lamprey spinal cord. It was shown that (1) all studied EAA (glutamate, kainate, NMDA, aspartate, and quisqualate) evoke an ion current and a short-term reversible depolarization in studied cells; (2) EAA added to perfusion solution may produce MP oscillations, with kinetic parameters and duration of the oscillation depending on the amino acid used (the most effective are kainate and NMDA, the least effective, quisqualate); (3) oscillations can be irregular (of the type of a synaptic noise or of a long-term plateau of depolarization with action potentials—AP) or regular, with frequency of 0.5–1.5 Hz. Amplitude of both oscillation types depends on MP level, frequency is more steady for each cell and less depends on MP. In 68 out of 128 studied cells, oscillations could be evoked, which indicates that a significant part of lamprey spinal neurons have intrinsic capability for MP oscillations and probably pacemaker properties. The functional role of oscillations can be different. They can take cells out from the profound inhibition state, synchronize activity of rhythm generation neurons and/or be the base for trigger signals (AP firing) sent by locomotor neuronal circuits to trunk muscles.     

Convergence of preganglionic fibers on superior cervical ganglion neurons in the rabbit

Intracellular recording techniques were used to record electrical response from neurons of the rabbit (isolated) superior cervical ganglion to single stimuli applied to bundles of preganglionic fibers as well as tonic electrical neuronal activity in this ganglion during acute experiments in situ. A review of the findings obtained confirms that neurons of the ganglion receive preganglionic synaptic inputs of two types, the first of a single pattern, formed by a preganglionic fiber; excitatory action of the latter on ganglion units suffices to induce postsynaptic action potentials (AP I) and the second of a multiple pattern, formed by several preganglionic fibers with relatively faint excitatory action, capable of evoking postsynaptic action potentials (AP II) only when excited in unison. Interspike intervals for AP I and AP II in tonic neuronal activity conformed to a normal distribution and a random distribution pattern respectively.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 21, No. 2, pp. 252–261, March–April, 1989.     

AMPA receptor trafficking at excitatory synapses

Excitatory synapses in the CNS release glutamate, which acts primarily on two sides of ionotropic receptors: AMPA receptors and NMDA receptors. AMPA receptors mediate the postsynaptic depolarization that initiates neuronal firing, whereas NMDA receptors initiate synaptic plasticity. Recent studies have emphasized that distinct mechanisms control synaptic expression of these two receptor classes. Whereas NMDA receptor proteins are relatively fixed, AMPA receptors cycle synaptic membranes on and off. A large family of interacting proteins regulates AMPA receptor turnover at synapses and thereby influences synaptic strength. Furthermore, neuronal activity controls synaptic AMPA receptor trafficking, and this dynamic process plays a key role in the synaptic plasticity that is thought to underlie aspects of learning and memory.