A serial multiplex immunogold labeling method for identifying peptidergic 
neurons in connectomes

Shahidi, R; Williams, EA; Conzelmann, M; Asadulina, A; Verasztó, C; Jasek, S; Bezares-Calderón, LA; Jékely, G

doi:10.7554/eLife.11147

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A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes

MPS-Authors

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Shahidi, R
Research Group Neurobiology of Marine Zooplankton, Max Planck Institute for Developmental Biology, Max Planck Society;

/persons/resource/persons273025

Williams, EA
Research Group Neurobiology of Marine Zooplankton, Max Planck Institute for Developmental Biology, Max Planck Society;

/persons/resource/persons273027

Conzelmann, M
Research Group Neurobiology of Marine Zooplankton, Max Planck Institute for Developmental Biology, Max Planck Society;

/persons/resource/persons274971

Asadulina, A
Research Group Neurobiology of Marine Zooplankton, Max Planck Institute for Developmental Biology, Max Planck Society;

/persons/resource/persons84766

Verasztó, C
Research Group Neurobiology of Marine Zooplankton, Max Planck Institute for Developmental Biology, Max Planck Society;

/persons/resource/persons273016

Jasek, S
Research Group Neurobiology of Marine Zooplankton, Max Planck Institute for Developmental Biology, Max Planck Society;

/persons/resource/persons273014

Bezares-Calderón, LA
Research Group Neurobiology of Marine Zooplankton, Max Planck Institute for Developmental Biology, Max Planck Society;

/persons/resource/persons85269

Jékely, G
Research Group Neurobiology of Marine Zooplankton, Max Planck Institute for Developmental Biology, Max Planck Society;

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引用

Shahidi, R., Williams, E., Conzelmann, M., Asadulina, A., Verasztó, C., Jasek, S., Bezares-Calderón, L., & Jékely, G. (2015). A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes. eLife, 4:. doi:10.7554/eLife.11147.

引用: https://hdl.handle.net/21.11116/0000-000A-9470-0

要旨

Electron microscopy-based connectomics aims to comprehensively map synaptic connections in neural tissue. However, current approaches are limited in their capacity to directly assign molecular identities to neurons. Here, we use serial multiplex immunogold labeling (siGOLD) and serial-section transmission electron microscopy (ssTEM) to identify multiple peptidergic neurons in a connectome. The high immunogenicity of neuropeptides and their broad distribution along axons, allowed us to identify distinct neurons by immunolabeling small subsets of sections within larger series. We demonstrate the scalability of siGOLD by using 11 neuropeptide antibodies on a full-body larval ssTEM dataset of the annelid Platynereis. We also reconstruct a peptidergic circuitry comprising the sensory nuchal organs, found by siGOLD to express pigment-dispersing factor, a circadian neuropeptide. Our approach enables the direct overlaying of chemical neuromodulatory maps onto synaptic connectomic maps in the study of nervous systems.