RESUMO
Over a century ago, the Spanish anatomist Ramón y Cajal described 'mossy fibres' in the hippocampus and the cerebellum, which contain several presynaptic boutons. Technical improvements in recent decades have allowed direct patch-clamp recordings from both hippocampal and cerebellar mossy fibre boutons (hMFBs and cMFBs, respectively), making them ideal models to study fundamental properties of synaptic transmission. hMFBs and cMFBs have similar size and shape, but each hMFB contacts one postsynaptic hippocampal CA3 pyramidal neuron, while each cMFB contacts â¼50 cerebellar granule cells. Furthermore, hMFBs and cMFBs differ in terms of their functional specialization. At hMFBs, a large number of release-ready vesicles and low release probability (<0.1) contribute to marked synaptic facilitation. At cMFBs, a small number of release-ready vesicles, high release probability (â¼0.5) and rapid vesicle reloading result in moderate frequency-dependent synaptic depression. These presynaptic mechanisms, in combination with faster postsynaptic currents of cerebellar granule cells compared with hippocampal CA3 pyramidal neurons, enable much higher transmission frequencies at cMFB compared with hMFB synapses. Analysing the underling mechanisms of synaptic transmission and information processing represents a fascinating challenge and may reveal insights into the structure-function relationship of the human brain.
Assuntos
Fibras Musgosas Hipocampais/fisiologia , Fibras Nervosas/fisiologia , Animais , Plasticidade Neuronal , Transmissão SinápticaRESUMO
How can synapses change the amount of neurotransmitter released during synaptic plasticity? Although release in general is intensely investigated, its determinants during plasticity are still poorly understood. As a model for plastic strengthening of synaptic release, we here use the well-established presynaptic homeostatic compensation during interference with postsynaptic glutamate receptors at the Drosophila neuromuscular junction. Combining short-term plasticity analysis, cumulative EPSC analysis, fluctuation analysis, and quantal short-term plasticity modeling, we found an increase in the number of release-ready vesicles during presynaptic strengthening. High-resolution light microscopy revealed an increase in the amount of the active zone protein Bruchpilot and an enlargement of the presynaptic cytomatrix structure. Furthermore, these functional and structural alterations of the active zone were not only observed after lifelong but already after minutes of presynaptic strengthening. Our results demonstrate that presynaptic plasticity can induce active zone remodeling, which regulates the number of release-ready vesicles within minutes.
Assuntos
Plasticidade Neuronal/fisiologia , Sinapses/fisiologia , Transmissão Sináptica/fisiologia , Vesículas Sinápticas/fisiologia , Animais , Animais Geneticamente Modificados , Drosophila , Eletrofisiologia , Potenciais Pós-Sinápticos Excitadores/fisiologia , Modelos Neurológicos , Receptores de Glutamato/metabolismoRESUMO
The synapse-associated protein of 47 kDa (SAP47) is a member of a phylogenetically conserved gene family of hitherto unknown function. In Drosophila, SAP47 is encoded by a single gene (Sap47) and is expressed throughout all synaptic regions of the wild-type larval brain; specifically, electron microscopy reveals anti-SAP47 immunogold labeling within 30 nm of presynaptic vesicles. To analyze SAP47 function, we used the viable and fertile deletion mutant Sap47(156), which suffers from a 1.7 kb deletion in the regulatory region and the first exon. SAP47 cannot be detected by either immunoblotting or immunohistochemistry in Sap47(156) mutants. These mutants exhibit normal sensory detection of odorants and tastants as well as normal motor performance and basic neurotransmission at the neuromuscular junction. However, short-term plasticity at this synapse is distorted. Interestingly, Sap47(156) mutant larvae also show a 50% reduction in odorant-tastant associative learning ability; a similar associative impairment is observed in a second deletion allele (Sap47(201)) and upon reduction of SAP47 levels using RNA interference. In turn, transgenically restoring SAP47 in Sap47(156) mutant larvae rescues the defect in associative function. This report thus is the first to suggest a function for SAP47. It specifically argues that SAP47 is required for proper behavioral and synaptic plasticity in flies-and prompts the question whether its homologs are required for proper behavioral and synaptic plasticity in other species as well.
Assuntos
Proteínas de Drosophila/deficiência , Atividade Motora/fisiologia , Proteínas do Tecido Nervoso/deficiência , Plasticidade Neuronal/fisiologia , Sinapses/metabolismo , Animais , Animais Geneticamente Modificados , Proteínas de Drosophila/genética , Drosophila melanogaster , Técnicas de Silenciamento de Genes , Masculino , Proteínas do Tecido Nervoso/genética , Olfato/fisiologia , Sinapses/genéticaRESUMO
At presynaptic active zones (AZs), the frequently observed tethering of synaptic vesicles to an electron-dense cytomatrix represents a process of largely unknown functional significance. Here, we identified a hypomorphic allele, brpnude, lacking merely the last 1% of the C-terminal amino acids (17 of 1740) of the active zone protein Bruchpilot. In brpnude, electron-dense bodies were properly shaped, though entirely bare of synaptic vesicles. While basal glutamate release was unchanged, paired-pulse and sustained stimulation provoked depression. Furthermore, rapid recovery following sustained release was slowed. Our results causally link, with intramolecular precision, the tethering of vesicles at the AZ cytomatrix to synaptic depression.
Assuntos
Proteínas de Drosophila/genética , Terminações Pré-Sinápticas/fisiologia , Vesículas Sinápticas/genética , Vesículas Sinápticas/fisiologia , Animais , Sequência de Bases , Canais de Cálcio/metabolismo , Citoplasma/metabolismo , Drosophila , Estimulação Elétrica , Eletrofisiologia , Cinética , Larva , Microscopia Eletrônica , Dados de Sequência Molecular , Mutação , Técnicas de Patch-ClampRESUMO
BMP signaling responses are refined by distinct secreted and intracellular antagonists in different cellular and temporal contexts. Here, we show that the nuclear LEM-domain protein MAN1 is a tissue-specific antagonist of BMP signaling in Drosophila. MAN1 contains two potential Mad-binding sites. We generated MAN1DeltaC mutants, harbouring a MAN1 protein that lacks part of the C-terminus including the RNA recognition motif, a putative Mad-binding domain. MAN1DeltaC mutants show wing crossvein (CV) patterning defects but no detectable alterations in nuclear morphology. MAN1(DeltaC) pupal wings display expanded phospho-Mad (pMad) accumulation and ectopic expression of the BMP-responsive gene crossveinless-2 (cv-2) indicating that MAN1 restricts BMP signaling. Conversely, MAN1 overexpression in wing imaginal discs inhibited crossvein development and BMP signaling responses. MAN1 is expressed at high levels in pupal wing veins and can be activated in intervein regions by ectopic BMP signaling. The specific upregulation of MAN1 in pupal wing veins may thus represent a negative feedback circuit that limits BMP signaling during CV formation. MAN1DeltaC flies also show reduced locomotor activity, and electrophysiology recordings in MAN1DeltaC larvae uncover a new presynaptic role of MAN1 at the neuromuscular junction (NMJ). Genetic interaction experiments suggest that MAN1 is a BMP signaling antagonist both at the NMJ and during CV formation.
Assuntos
Proteínas Morfogenéticas Ósseas/metabolismo , Proteínas de Drosophila/fisiologia , Drosophila/embriologia , Junção Neuromuscular/embriologia , Proteínas Nucleares/fisiologia , Transdução de Sinais/fisiologia , Asas de Animais/embriologia , Animais , Sequência de Bases , Sítios de Ligação , Primers do DNA , Proteínas de Drosophila/metabolismo , Eletroforese em Gel de Poliacrilamida , Imuno-Histoquímica , Imunoprecipitação , Proteínas Nucleares/metabolismoRESUMO
Sustained rate-coded signals encode many types of sensory modalities. Some sensory synapses possess specialized ribbon structures, which tether vesicles, to enable high-frequency signaling. However, central synapses lack these structures, yet some can maintain signaling over a wide bandwidth. To analyze the underlying molecular mechanisms, we investigated the function of the active zone core component Bassoon in cerebellar mossy fiber to granule cell synapses. We show that short-term synaptic depression is enhanced in Bassoon knockout mice during sustained high-frequency trains but basal synaptic transmission is unaffected. Fluctuation and quantal analysis as well as quantification with constrained short-term plasticity models revealed that the vesicle reloading rate was halved in the absence of Bassoon. Thus, our data show that the cytomatrix protein Bassoon speeds the reloading of vesicles to release sites at a central excitatory synapse.