Depression Of Synaptic Transmission Research Articles

Repetitive transcranial magnetic stimulation (rTMS) triggers dose-dependent homeostatic rewiring in recurrent neuronal networks.

Repetitive transcranial magnetic stimulation (rTMS) is a non-invasive brain stimulation technique used to induce neuronal plasticity in healthy individuals and patients. Designing effective and reproducible rTMS protocols poses a major challenge in the field as the underlying biomechanisms of long-term effects remain elusive. Current clinical protocol designs are often based on studies reporting rTMS-induced long-term potentiation or depression of synaptic transmission. Herein, we employed computational modeling to explore the effects of rTMS on long-term structural plasticity and changes in network connectivity. We simulated a recurrent neuronal network with homeostatic structural plasticity among excitatory neurons, and demonstrated that this mechanism was sensitive to specific parameters of the stimulation protocol (i.e., frequency, intensity, and duration of stimulation). Particularly, the feedback-inhibition initiated by network stimulation influenced the net stimulation outcome and hindered the rTMS-induced structural reorganization, highlighting the role of inhibitory networks. These findings suggest a novel mechanism for the lasting effects of rTMS, i.e., rTMS-induced homeostatic structural plasticity, and highlight the importance of network inhibition in careful protocol design, standardization, and optimization of stimulation.

A Common Human Brain-Derived Neurotrophic Factor Polymorphism Leads to Prolonged Depression of Excitatory Synaptic Transmission by Isoflurane in Hippocampal Cultures.

Multiple presynaptic and postsynaptic targets have been identified for the reversible neurophysiological effects of general anesthetics on synaptic transmission and neuronal excitability. However, the synaptic mechanisms involved in persistent depression of synaptic transmission resulting in more prolonged neurological dysfunction following anesthesia are less clear. Here, we show that brain-derived neurotrophic factor (BDNF), a growth factor implicated in synaptic plasticity and dysfunction, enhances glutamate synaptic vesicle exocytosis, and that attenuation of vesicular BDNF release by isoflurane contributes to transient depression of excitatory synaptic transmission in mice. This reduction in synaptic vesicle exocytosis by isoflurane was acutely irreversible in neurons that release less endogenous BDNF due to a polymorphism (BDNF Val66Met; rs6265) compared to neurons from wild-type mice. These effects were prevented by exogenous application of BDNF. Our findings identify a role for a common human BDNF single nucleotide polymorphism in persistent changes of synaptic function following isoflurane exposure. These short-term persistent alterations in excitatory synaptic transmission indicate a role for human genetic variation in anesthetic effects on synaptic plasticity and neurocognitive function.

The effects of tricaine mesylate on arthropods: crayfish, crab and Drosophila.

Tricaine mesylate, also known as MS-222, was investigated to characterize its effects on sensory neurons, synaptic transmission at the neuromuscular junction, and heart rate in invertebrates. Three species were examined: Drosophila melanogaster, blue crab (Callinectes sapidus), and red swamp crayfish (Procambarus clarkii). Intracellular measures of action potentials in motor neurons of the crayfish demonstrated that MS-222 dampened the amplitude, suggesting that voltage-gated Na + channels are blocked by MS-222. This is likely the mechanism behind the reduced activity measured in sensory neurons and depressed synaptic transmission in all three species as well as reduced cardiac function in the larval Drosophila. To address public access to data, a group effort was used for analysis of given data sets, blind to the experimental design, to gauge analytical accuracy. The determination of a threshold in analysis for measuring extracellular recorded sensory events is critical and is not easily performed with commercial software.

Activation of α-adrenoceptors depresses synaptic transmission of myelinated afferents and inhibits pathways mediating primary afferent depolarization (PAD) in the in vitro mouse spinal cord.

Somatosensory afferent transmission strength is controlled by several presynaptic mechanisms that reduce transmitter release at the spinal cord level. We focused this investigation on the role of α-adrenoceptors in modulating sensory transmission in low-threshold myelinated afferents and in pathways mediating primary afferent depolarization (PAD) of neonatal mouse spinal cord. We hypothesized that the activation of α-adrenoceptors depresses low threshold-evoked synaptic transmission and inhibits pathways mediating PAD. Extracellular field potentials (EFPs) recorded in the deep dorsal horn assessed adrenergic modulation of population monosynaptic transmission, while dorsal root potentials (DRPs) recorded at root entry zone assessed adrenergic modulation of PAD. We found that noradrenaline (NA) and the α1-adrenoceptor agonists phenylephrine and cirazoline depressed synaptic transmission (by 15, 14 and 22%, respectively). DRPs were also depressed by NA, phenylephrine and cirazoline (by 62, 30, and 64%, respectively), and by the α2-adrenoceptor agonist clonidine, although to a lower extent (20%). We conclude that NA depresses monosynaptic transmission of myelinated afferents onto deep dorsal horn neurons via α1-adrenoceptors and inhibits interneuronal pathways mediating PAD through the activation of α1- and α2-adrenoceptors. The functional significance of these modulatory actions in shaping cutaneous and muscle sensory information during motor behaviors requires further study.

PSD-95 Serine 73 phosphorylation is not required for induction of NMDA-LTD

PSD-95 is a major scaffolding protein of the post-synaptic density (PSD) of a glutamatergic synapse. PSD-95, via interactions with stargazin, anchors AMPA receptors at the synapse and regulates AMPAR currents. The expression of PSD-95 is regulated during synaptic plasticity. It is, however, unknown whether this regulation is required for induction of functional plasticity of glutamatergic synapses. Here, we show that NMDA-induced long-term depression of synaptic transmission (NMDA-LTD) is accompanied by downregulation of PSD-95 protein levels. Using pharmacologic and molecular manipulations, we further demonstrate that the NMDA-induced downregulation of PSD-95 depends on the activation of CaMKII and CaMKII-driven phosphorylation of PSD-95 serine 73. Surprisingly, neither CaMKII activity nor CaMKII-dependent phosphorylation of PSD-95 serine 73 are required for the expression of NMDA-LTD. These results support the hypothesis that synaptic plasticity of AMPARs may occur without dynamic regulation of PSD-95 protein levels.

ЗАКОНОМІРНОСТІ СПОНТАННОГО КВАНТОВОГО ВИВІЛЬНЕННЯ ГЛУТАМАТУ В СИНАПСАХ НЕЙРОНІВ ПЕРВИННОЇ КУЛЬТУРИ ГІПОКАМПА ПРИ ДОВГОТРИВАЛІЙ ДЕПРЕСІЇ СИНАПТИЧНОЇ ПЕРЕДАЧІ

The results of electrophysiological studies of the quantal release features of glutamate in hippocampal neurons' synapses in long-term depression of synaptic transmission are presented. To research the topic, weused the primary culture of the rat hippocampus. Using the patch-clamp technique, the frequencies and amplitudes of excitatory spontaneous postsynaptic currents (EPSCs) were determined, theirdistributions were analyzed and the basicquantal parameters were calculated. Long-term depression of synaptic transmission was achieved by tetanic stimulation of the axon of the presynaptic cell for 5 minutes with a stimulation frequency of 5 Hz. Spontaneous current in depression was recorded 20-30 min after tetanic stimulation. It has been shown that the amplitude of the EPSCs registered in hippocampal neurons decreased in the long-term depression of synaptic transmission. It has been found that two neurotransmitter vesicles are usually released in the control at the same time and one during the long-term depression of synaptic transmission.It means that the probability of release decreased. In addition, the average quantalamplitude and quantum content decreased. It indicates the effect of presynaptic mechanisms in the expression of long-term depression of synaptic neurotransmission between hippocampal neurons in culture.In the conditions of long-term depression, no significant changes were found in the frequency of EPSCs. The analysis of quantum parameters in long-term depression of glutamatergic synaptic transmission between neurons is important for the formation of more complete ideas about the mechanisms that play a fundamental role in the normal functioning of the CNS and the development of neural networks.

Depressed Synaptic Transmission and Reduced Vesicle Release Sites in Huntington's Disease Neuromuscular Junctions

Huntington's disease (HD) is a progressive and fatal degenerative disorder that results in debilitating cognitive and motor dysfunction. Most HD studies have focused on degeneration of the CNS. We previously discovered that skeletal muscle from transgenic R6/2 HD mice is hyperexcitable due to decreased chloride and potassium conductances. The progressive and early onset of these defects suggest a primary myopathy in HD. In this study, we examined the relationship between neuromuscular transmission and skeletal muscle hyperexcitability. We used an ex vivo preparation of the levator auris longus muscle from male and female late-stage R6/2 mice and age-matched wild-type controls. Immunostaining of the synapses and molecular analyses revealed no evidence of denervation. Physiologically, we recorded spontaneous miniature endplate currents (mEPCs) and nerve-evoked EPCs (eEPCs) under voltage-clamp, which, unlike current-clamp records, were independent of the changes in muscle membrane properties. We found a reduction in the number of vesicles released per action potential (quantal content) in R6/2 muscle, which analysis of eEPC variance and morphology indicate is caused by a reduction in the number of vesicle release sites (n) rather than a change in the probability of release (prel). Furthermore, analysis of high-frequency stimulation trains suggests an impairment in vesicle mobilization. The depressed neuromuscular transmission in R6/2 muscle may help compensate for the muscle hyperexcitability and contribute to motor impersistence.SIGNIFICANCE STATEMENT Recent evidence indicates that Huntington's disease (HD) is a multisystem disorder. Our examination of neuromuscular transmission in this study reveals defects in the motor nerve terminal that may compensate for the muscle hyperexcitability in HD. The technique we used eliminates the effects of the altered muscle membrane properties on synaptic currents and thus provides hitherto the most detailed analysis of synaptic transmission in HD. Clinically, the striking depression of neurotransmission we found may help explain the motor impersistence in HD patients. Therapies that target the highly accessible peripheral nerve and muscle system provide a promising new avenue to lessen the debilitating motor symptoms of HD.

Mechanisms of Long-Term Plasticity of Hippocampal GABAergic Synapses.

Long-term potentiation and depression of synaptic transmission have been considered as cellular mechanisms of memory in studies conducted in recent decades. These studies were predominantly focused on mechanisms underlying plasticity at excitatory synapses. Nevertheless, normal central nervous system functioning requires maintenance of a balance between inhibition and excitation, suggesting existence of similar modulation of glutamatergic and GABAergic synapses. Here we review the involvement of G-protein-coupled receptors in the generation of long-term changes in synaptic transmission of inhibitory synapses. We considered the role of endocannabinoid and glutamate systems, GABAB and opioid receptors in the induction of long-term potentiation and long-term depression in inhibitory synapses. The pre- and postsynaptic effects of activation of these receptors are also discussed.

Herpes Simplex Virus type-1 infection induces synaptic dysfunction in cultured cortical neurons via GSK-3 activation and intraneuronal amyloid-β protein accumulation.

Increasing evidence suggests that recurrent Herpes Simplex Virus type 1 (HSV-1) infection spreading to the CNS is a risk factor for Alzheimer’s Disease (AD) but the underlying mechanisms have not been fully elucidated yet. Here we demonstrate that in cultured mouse cortical neurons HSV-1 induced Ca2+-dependent activation of glycogen synthase kinase (GSK)-3. This event was critical for the HSV-1-dependent phosphorylation of amyloid precursor protein (APP) at Thr668 and the following intraneuronal accumulation of amyloid-β protein (Aβ). HSV-1-infected neurons also exhibited: i) significantly reduced expression of the presynaptic proteins synapsin-1 and synaptophysin; ii) depressed synaptic transmission. These effects depended on GSK-3 activation and intraneuronal accumulation of Aβ. In fact, either the selective GSK-3 inhibitor, SB216763, or a specific antibody recognizing Aβ (4G8) significantly counteracted the effects induced by HSV-1 at the synaptic level. Moreover, in neurons derived from APP KO mice and infected with HSV-1 Aβ accumulation was not found and synaptic protein expression was only slightly reduced when compared to wild-type infected neurons. These data further support our contention that HSV-1 infections spreading to the CNS may contribute to AD phenotype.

Localization of Presynaptic Plasticity Mechanisms Enables Functional Independence of Synaptic and Ectopic Transmission in the Cerebellum.

In the cerebellar molecular layer parallel fibre terminals release glutamate from both the active zone and from extrasynaptic “ectopic” sites. Ectopic release mediates transmission to the Bergmann glia that ensheathe the synapse, activating Ca2+-permeable AMPA receptors and glutamate transporters. Parallel fibre terminals exhibit several forms of presynaptic plasticity, including cAMP-dependent long-term potentiation and endocannabinoid-dependent long-term depression, but it is not known whether these presynaptic forms of long-term plasticity also influence ectopic transmission to Bergmann glia. Stimulation of parallel fibre inputs at 16 Hz evoked LTP of synaptic transmission, but LTD of ectopic transmission. Pharmacological activation of adenylyl cyclase by forskolin caused LTP at Purkinje neurons, but only transient potentiation at Bergmann glia, reinforcing the concept that ectopic sites lack the capacity to express sustained cAMP-dependent potentiation. Activation of mGluR1 caused depression of synaptic transmission via retrograde endocannabinoid signalling but had no significant effect at ectopic sites. In contrast, activation of NMDA receptors suppressed both synaptic and ectopic transmission. The results suggest that the signalling mechanisms for presynaptic LTP and retrograde depression by endocannabinoids are restricted to the active zone at parallel fibre synapses, allowing independent modulation of synaptic transmission to Purkinje neurons and ectopic transmission to Bergmann glia.

Short-Term Plasticity of Glutamatergic and GABA-Ergic Synaptic Transmission between Co-Cultured Retinal Ganglion Cells and Superior Colliculus Neurons

Received 21 November, 2013. We studied changes in the efficacy of synaptic transmission via glutamatergic and GABA-ergic synaptic connections between co-cultured retinal ganglion cells (RGCs) and neurons of the superior colliculus (SC) at paired stimulation of RGCs (as a form of short-term synaptic plasticity). In a parallel manner, we recorded transmembrane currents and potentials in synaptically connected pairs of RGCs and SC neurons using the paired whole-cell patch-clamp technique. When glutamatergic synaptic action was mediated by activation of exclusively NMDA or non-NMDA receptor channel complexes on the postsynaptic membrane of SC neurons, depression of synaptic transmission after the second AP was observed. In the case of GABA-ergic transmission mediated by activation of GABA А receptor-channel complexes, analogous paired stimulation resulted in facilitation of synaptic action on an SC neuron after the second AP generated by an RGC. The use of basic and complete quantal analyses allowed us to find a significant decrease (Р < 0.05) in the binomial parameter n in the case of depression of NMDA-mediated evoked postsynaptic currents (ePSCs) and a significant drop in both quantal parameter q and binomial parameters n and p at depression of non-NMDA-mediated ePSCs. In the former case the estimate was indicative of possible presynaptic localization of the mechanisms of depression resulting from a decrease in the number of released synaptic vesicles containing the transmitter, while in the latter case not only pre- but also post-synaptic mechanisms (decrease in the number of released synaptic vesicles and desensitization of postsynaptic receptors) are probably involved. Estimation of normalized alterations of the quantal and binomial parameters at facilitation of GABA А -mediated ePSCs demonstrated a significant increase (Р < 0.05) in the presynaptic factors n and p and, correspondingly, in the quantal content m. Therefore, the effect of facilitation can be due to the processes realized in a presynaptic terminal (enhancement of the number of synaptic vesicles).

APP Homodimers Transduce an Amyloid-β-Mediated Increase in Release Probability at Excitatory Synapses

Accumulation of amyloid-β peptides (Aβ), the proteolytic products of the amyloid precursor protein (APP), induces a variety of synaptic dysfunctions ranging from hyperactivity to depression that are thought to cause cognitive decline in Alzheimer's disease. While depression of synaptic transmission has been extensively studied, the mechanisms underlying synaptic hyperactivity remain unknown. Here, we show that Aβ40 monomers and dimers augment release probability through local fine-tuning of APP-APP interactions at excitatory hippocampal boutons. Aβ40 binds to the APP, increases the APP homodimer fraction at the plasma membrane, and promotes APP-APP interactions. The APP activation induces structural rearrangements in the APP/Gi/o-protein complex, boosting presynaptic calcium flux and vesicle release. The APP growth-factor-like domain (GFLD) mediates APP-APP conformational changes and presynaptic enhancement. Thus, the APP homodimer constitutes a presynaptic receptor that transduces signal from Aβ40 to glutamate release. Excessive APP activation may initiate a positive feedback loop, contributing to hippocampal hyperactivity in Alzheimer's disease.

ProBDNF Negatively Regulates Neuronal Remodeling, Synaptic Transmission, and Synaptic Plasticity in Hippocampus

Experience-dependent plasticity shapes postnatal development of neural circuits, but the mechanisms that refine dendritic arbors, remodel spines, and impair synaptic activity are poorly understood. Mature brain-derived neurotrophic factor (BDNF) modulates neuronal morphology and synaptic plasticity, including long-term potentiation (LTP) via TrkB activation. BDNF is initially translated as proBDNF, which binds p75(NTR). Invitro, recombinant proBDNF modulates neuronal structure and alters hippocampal long-term plasticity, but the actions ofendogenously expressed proBDNF are unclear. Therefore, we generated a cleavage-resistant probdnf knockin mouse. Our results demonstrate that proBDNF negatively regulates hippocampal dendritic complexity and spine density through p75(NTR). Hippocampal slices from probdnf mice exhibit depressed synaptic transmission, impaired LTP, and enhanced long-term depression (LTD) in area CA1. These results suggest that proBDNF acts invivo as a biologically active factor that regulates hippocampal structure, synaptic transmission, and plasticity, effects that are distinct from those of mature BDNF.

The Glutamatergic Synaptic Transmission Mediated by Ionotropic Glutamate Receptors in Rats Cerebral Cortex in Behavioral Depression

In experiments on rats was simulated the behavioral depression caused by social isolation and chronic inflammation. It was found in slices of the medial prefrontal cortex that behavioral depression was attained by the inhibition of the basal glutamatergic synaptic transmission in the layer V pyramidal neurons of the medial prefrontal cortex, that was manifested by decay of the amplitude of field (f) excitatory postsynaptic potentials (EPSPs). The increase of amlitude of N-methyl-D-aspartate (NMDA) component of fEPSPs in addition observed. Last effect was conditioned by the increase of the functional activity of the NMDA receptors (NMDARs) containing NR2A subunit. The increase of functional activity of the NMDARs evoked inhibition of the basal glutamatergic synaptic transmission in the pyramidal neurons, whereas it was prevented by systemic administration of NMDARs blocker ketamine. It was observed also under behavioral depression disturbances of the plastic properties of cortical synapses, which become apparent by the inhibition of the expression of long-term potentiation and long-term depression of synaptic transmission, but with the reinforcement of the last under the induced by social isolation of behavioral depression. The disturbances of synaptic plasticity are conditioned par excellence by increase of functional activity of verapamil-sensetive Ca 2+ channels.

Long‐term depression of synaptic transmission in the adult mouse insular cortex in vitro

The insular cortex (IC) is known to play important roles in higher brain functions such as memory and pain. Activity-dependent long-term depression (LTD) is a major form of synaptic plasticity related to memory and chronic pain. Previous studies of LTD have mainly focused on the hippocampus, and no study in the IC has been reported. In this study, using a 64-channel recording system, we show for the first time that repetitive low-frequency stimulation (LFS) can elicit frequency-dependent LTD of glutamate receptor-mediated excitatory synaptic transmission in both superficial and deep layers of the IC of adult mice. The induction of LTD in the IC required activation of the N-methyl-d-aspartate (NMDA) receptor, metabotropic glutamate receptor (mGluR)5, and L-type voltage-gated calcium channel. Protein phosphatase 1/2A and endocannabinoid signaling are also critical for the induction of LTD. In contrast, inhibiting protein kinase C, protein kinase A, protein kinase Mζ or calcium/calmodulin-dependent protein kinase II did not affect LFS-evoked LTD in the IC. Bath application of the group I mGluR agonist (RS)-3,5-dihydroxyphenylglycine produced another form of LTD in the IC, which was NMDA receptor-independent and could not be occluded by LFS-induced LTD. Our studies have characterised the basic mechanisms of LTD in the IC at the network level, and suggest that two different forms of LTD may co-exist in the same population of IC synapses.

Neurochemical Nature of Cognitive and Emotional Disorders under Conditions of Experimental Diabetes Mellitus

Experimental diabetes mellitus (EDM) was induced in rats by injections of alloxan (125 mg/kg); this disease was manifested in a rise in the mean level of glucose in the blood from 5.0 to 18.7 mM. On the 4th week of the development of EDM, rats demonstrated increased levels of depression and anxiety and worsening of the processes of formation and reproduction of trained skills. Behavioral modifications were developed against the background of decreases in the amplitudes of NMDA components of field EPSPs (fEPSPs) of pyramidal neurons of the hippocampus and prefrontal neocortex, weakening of the expression of long-term potentiation, and, concurrently, intensified development of long-term depression of synaptic transmission. Injections of insulin (durable action, 2 U per animal) during 2 days resulted in a decrease in the level of glucose in the blood but did not change behavioral and neurochemical disorders typical of EDM. The action of glucose in a high concentration (30 mM) on cerebral slices obtained from intact rats did not influence the relay and plastic properties of glutamatergic synapses of neurons of the hippocampus and neocortex; the action of 0.1 U/ml insulin under these conditions led to an increase in the amplitude of NMDA EPSPcomponents and facilitated the development of plastic phenomena. A blocker of calcium L-type channels, verapamil (20 mg/kg), exerted no significant effect on the relay and plastic properties of synapses between hippocampal and neocortical neurons but partially weakened disorders of behavior and synaptic plasticity under EDM conditions. Such disorders were eliminated by chronic (2 weeks) injections of antidepressants, imipramine and fluoxetine (doses 20 mg/kg), against the background of clearly pronounced hyperglycemia and weakening of functional activity of neuronal NMDA receptors. We hypothesize that a deficiency of activation of insulin receptors of cerebral neurons and hyperglycemia cause a rise in the level of corticosteroids in the blood and brain, and this disturbs synaptic plasticity and behavior. This supposition is based on the ability of calcium channel blockers and (especially) antidepressants to reduce behavioral and neurochemical abnormalities in EDM.

Reduced d-serine levels in the nucleus accumbens of cocaine-treated rats hinder the induction of NMDA receptor-dependent synaptic plasticity

Cocaine seeking behaviour and relapse have been linked to impaired potentiation and depression at excitatory synapses in the nucleus accumbens, but the mechanism underlying this process is poorly understood. We show that, in the rat nucleus accumbens core, D-serine is the endogenous coagonist of N-methyl-D-aspartate receptors, and its presence is essential for N-methyl-D-aspartate receptor-dependent potentiation and depression of synaptic transmission. Nucleus accumbens core slices obtained from cocaine-treated rats after 1 day of abstinence presented significantly reduced D-serine concentrations, increased expression of the D-serine degrading enzyme, D-amino acid oxidase, and downregulated expression of serine racemase, the enzyme responsible for D-serine synthesis. The D-serine deficit was associated with impairment of potentiation and depression of glutamatergic synaptic transmission, which was restored by slice perfusion with exogenous D-serine. Furthermore, in vivo administration of D-serine directly into the nucleus accumbens core blocked behavioural sensitization to cocaine. These results provide evidence for a critical role of D-serine signalling in synaptic plasticity relevant to cocaine addiction.

Impairment of synaptic plasticity by the stress mediator CRH involves selective destruction of thin dendritic spines via RhoA signaling

Stress is ubiquitous in modern life and exerts profound effects on cognitive and emotional functions. Thus, whereas acute stress enhances memory, longer episodes exert negative effects through as yet unresolved mechanisms. We report a novel, hippocampus-intrinsic mechanism for the selective memory defects that are provoked by stress. CRH (corticotropin-releasing hormone), a peptide released from hippocampal neurons during stress, depressed synaptic transmission, blocked activity-induced polymerization of spine actin and impaired synaptic plasticity in adult hippocampal slices. Live, multiphoton imaging demonstrated a selective vulnerability of thin dendritic spines to this stress hormone, resulting in depletion of small, potentiation-ready excitatory synapses. The underlying molecular mechanisms required activation and signaling of the actin-regulating small GTPase, RhoA. These results implicate the selective loss of dendritic spine sub-populations as a novel structural and functional foundation for the clinically important effects of stress on cognitive and emotional processes.

Molecular Determinants of Modulation of CaV2.1 Channels by Visinin-like Protein 2

CaV2.1 channels, which conduct P/Q-type Ca2+ currents, initiate synaptic transmission at most synapses in the central nervous system. Ca2+/calmodulin-dependent facilitation and inactivation of these channels contributes to short-term facilitation and depression of synaptic transmission, respectively. Other calcium sensor proteins displace calmodulin (CaM) from its binding site, differentially regulate CaV2.1 channels, and contribute to the diversity of short-term synaptic plasticity. The neuronal calcium sensor protein visinin-like protein 2 (VILIP-2) inhibits inactivation and enhances facilitation of CaV2.1 channels. Here we examine the molecular determinants for differential regulation of CaV2.1 channels by VILIP-2 and CaM by construction and functional analysis of chimeras in which the functional domains of VILIP-2 are substituted in CaM. Our results show that the N-terminal domain, including its myristoylation site, the central α-helix, and the C-terminal lobe containing EF-hands 3 and 4 of VILIP-2 are sufficient to transfer its regulatory properties to CaM. This regulation by VILIP-2 requires binding to the IQ-like domain of CaV2.1 channels. Our results identify the essential molecular determinants of differential regulation of CaV2.1 channels by VILIP-2 and define the molecular code that these proteins use to control short-term synaptic plasticity.

Molecular Determinants of CaV2.1 Channel Regulation by Calcium-binding Protein-1

Presynaptic Ca(V)2.1 channels, which conduct P/Q-type Ca(2+) currents, initiate synaptic transmission at most synapses in the central nervous system. Regulation of Ca(V)2.1 channels by CaM contributes significantly to short term facilitation and rapid depression of synaptic transmission. Short term synaptic plasticity is diverse in form and function at different synapses, yet CaM is ubiquitously expressed. Differential regulation of Ca(V)2.1 channels by CaM-like Ca(2+) sensor (CaS) proteins differentially affects short term synaptic facilitation and rapid synaptic depression in transfected sympathetic neuron synapses. Here, we define the molecular determinants for differential regulation of Ca(V)2.1 channels by the CaS protein calcium-binding protein-1 (CaBP1) by analysis of chimeras in which the unique structural domains of CaBP1 are inserted into CaM. Our results show that the N-terminal domain, including its myristoylation site, and the second EF-hand, which is inactive in Ca(2+) binding, are the key molecular determinants of differential regulation of Ca(V)2.1 channels by CaBP1. These findings give insight into the molecular code by which CaS proteins differentially regulate Ca(V)2.1 channel function and provide diversity of form and function of short term synaptic plasticity.