![]() ![]() This complex multi-step process is highly regulated and involves a large number of molecular components. During synaptic transmission, an action potential triggers the entry of calcium ions into the presynapse, which induces synaptic vesicle (SV) fusion and release of neurotransmitters into the synaptic cleft. They consist of pre- and post-synaptic compartments, whose function is to release and respond to neurotransmitters, respectively. Our software is applicable to a wide range of synaptic induction protocols (including spontaneous synaptogenesis observed in neuron cultures) and is a valuable tool to determine the subtle impact of disease-associated genes on presynaptic assembly.Ĭhemical synapses are specialized cellular junctions that permit information flow from one neuron to another. Our analysis revealed a novel role for miR-27b in augmenting the density of presynaptic clusters. Using this software, we assessed the intrinsic variability of this synaptic induction assay and probed the effect of eight neuronal microRNAs on presynaptic assembly. Here, we introduce an image-processing algorithm that identifies presynaptic clusters in mammalian co-cultures and extracts a range of synapse-specific parameters. The identification of regulatory pathways that fine-tune presynaptic assembly is hindered by the lack of adequate tools to quantitatively image this process. This assay has led to the discovery of numerous synaptogenic proteins, but has not been used to probe neuronal mechanisms regulating presynaptic induction. In mammalian neurons, presynaptic assembly is widely studied in a co-culture assay, where a synaptogenic cue expressed at the surface of a heterologous cell induces presynaptic differentiation in a contacting axon. ![]() Presynaptic assembly involves the specialization of a patch of axonal membrane into a complex structure that supports synaptic vesicle exocytosis and neurotransmitter release. 4Department of Physiology, National University of Singapore, Singapore, Singapore.3Department of Biochemistry, National University of Singapore, Singapore, Singapore.2Cancer and Stem Cell Biology Program, Duke-NUS Graduate Medical School, Singapore, Singapore.1Neuroscience and Behavioral Disorders Program, Duke-NUS Graduate Medical School, Singapore, Singapore.Mathijs Voorhoeve 2,3 and Marc Fivaz 1,4 * ![]()
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