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A Serial Multiplex Immunogold Labeling Method for Identifying Peptidergic Neurons in Connectomes

Reza Shahidi, Elizabeth Williams, Markus Conzelmann, Albina Asadulina, Csaba Veraszto, Sanja Jasek, Luis A Bezares-Calderon, Gaspar Jekely
doi: https://doi.org/10.1101/027458
Reza Shahidi
Max-Planck-Institute for Developmental Biology;
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Elizabeth Williams
Max-Planck-Institute for Developmental Biology;
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Markus Conzelmann
Max-Planck-Institute for Developmental Biology;
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Albina Asadulina
Max-Planck-Institute for Developmental Biology;
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Csaba Veraszto
Max-Planck-Institute for Developmental Biology;
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Sanja Jasek
Max-Planck-Institute for Developmental Biology;
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Luis A Bezares-Calderon
Max-Planck-Institute for Developmental Biology;
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Gaspar Jekely
Max Planck Institute for Developmental Biology
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  • For correspondence: gaspar.jekely@tuebingen.mpg.de
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Abstract

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.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted November 27, 2015.
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A Serial Multiplex Immunogold Labeling Method for Identifying Peptidergic Neurons in Connectomes
Reza Shahidi, Elizabeth Williams, Markus Conzelmann, Albina Asadulina, Csaba Veraszto, Sanja Jasek, Luis A Bezares-Calderon, Gaspar Jekely
bioRxiv 027458; doi: https://doi.org/10.1101/027458
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A Serial Multiplex Immunogold Labeling Method for Identifying Peptidergic Neurons in Connectomes
Reza Shahidi, Elizabeth Williams, Markus Conzelmann, Albina Asadulina, Csaba Veraszto, Sanja Jasek, Luis A Bezares-Calderon, Gaspar Jekely
bioRxiv 027458; doi: https://doi.org/10.1101/027458

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