A semi-correlative technique for the subcellular localization of proteins in Drosophila synapses

Jiao, Wei; Shupliakov, Andrey; Shupliakov, Oleg

doi:10.1016/j.jneumeth.2009.10.010

Cited by 11 publications

(9 citation statements)

References 26 publications

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“…Electron microscopy and ultrastructure quantification were previously described (Jiao et al., 2010). Only type Ib boutons, identified by postsynaptic subsynaptic reticulum structure, were selected for quantification.…”

Section: Methodsmentioning

confidence: 99%

Miniature Neurotransmission Regulates Drosophila Synaptic Structural Maturation

Choi

Imlach

Jiao

et al. 2014

Neuron

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SummaryMiniature neurotransmission is the transsynaptic process where single synaptic vesicles spontaneously released from presynaptic neurons induce miniature postsynaptic potentials. Since their discovery over 60 years ago, miniature events have been found at every chemical synapse studied. However, the in vivo necessity for these small-amplitude events has remained enigmatic. Here, we show that miniature neurotransmission is required for the normal structural maturation of Drosophila glutamatergic synapses in a developmental role that is not shared by evoked neurotransmission. Conversely, we find that increasing miniature events is sufficient to induce synaptic terminal growth. We show that miniature neurotransmission acts locally at terminals to regulate synapse maturation via a Trio guanine nucleotide exchange factor (GEF) and Rac1 GTPase molecular signaling pathway. Our results establish that miniature neurotransmission, a universal but often-overlooked feature of synapses, has unique and essential functions in vivo.

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Section: Methodsmentioning

confidence: 99%

Miniature Neurotransmission Regulates Drosophila Synaptic Structural Maturation

Choi

Imlach

Jiao

et al. 2014

Neuron

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“…For experiments, fillets were incubated in HL3 buffer containing either EDTA for 10 minutes, resting conditions, or stimulated by addition of 60 mM K + for 10 minutes. The specimens were fixed in 4% paraformaldehyde solution, in 0.1 M phosphate buffer, pH 7.2 and labeled and embedded for EM as earlier described (Jiao et al, 2010b). Serial ultrathin sections were cut with a diamond knife (Diatome), stained with 1% uranyl acetate and lead citrate on grids, and examined with a Tecnai 12 electron microscope (FEI).…”

Section: Preembedding Immunocytochemistry and Temmentioning

confidence: 99%

“…Membrane contours were traced using a digitizer and transferred into Maya 8.0H 3D-reconstruction program and surface rendered as earlier described (Jiao et al, 2010b).…”

Section: D Reconstruction Of Tem Imagesmentioning

confidence: 99%

The dynamin-binding domains of Dap160/Intersectin affect bulk membrane retrieval in synapses

Winther

Jiao

Vorontsova

et al. 2013

Journal of Cell Science

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SummaryDynamin-associated protein 160 kDa (Dap160)/intersectin interacts with several synaptic proteins and affects endocytosis and synapse development. The functional role of the different protein interaction domains is not well understood. Here we show that Drosophila Dap160 lacking the dynamin-binding SH3 domains does not affect the development of the neuromuscular junction but plays a key role in synaptic vesicle recycling. dap160 mutants lacking dynamin-interacting domains no longer accumulate dynamin properly at the periactive zone, and it becomes dispersed in the bouton during stimulation. This is accompanied by a reduction in uptake of the dye FM1-43 and an accumulation of large vesicles and membrane invaginations. However, we do not observe an increase in the number of clathrin-coated intermediates. We also note a depression in evoked excitatory junction potentials (EJPs) during high-rate stimulation, accompanied by aberrantly large miniature EJPs. The data reveal the important role of Dap160 in the targeting of dynamin to the periactive zone, where it is required to suppress bulk synaptic vesicle membrane retrieval during high-frequency activity.

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“…For conventional electron microscopy, fillets were fixed in 3% glutaraldehyde in 0.1 M PBS buffer (pH 7.2). Specimens were embedded in Durcupan and analyzed as described elsewhere (Jiao et al, 2010).…”

Section: Specimen Preparationmentioning

confidence: 99%