@article {Shahidi027458, author = {R{\'e}za Shahidi and Elizabeth Williams and Markus Conzelmann and Albina Asadulina and Csaba Veraszt{\'o} and Sanja Jasek and Luis A. Bezares-Calder{\'o}n and G{\'a}sp{\'a}r J{\'e}kely}, title = {siGOLD: a Serial Multiplex Immunogold Labeling Method for Identifying Peptidergic Neurons in Connectomes}, elocation-id = {027458}, year = {2015}, doi = {10.1101/027458}, publisher = {Cold Spring Harbor Laboratory}, abstract = {Connectomics aims to comprehensively map synaptic connections in blocks of neural tissue. However, current approaches are limited in directly assigning molecular identities to neurons in a connectome. Here, we use serial multiplex immunogold labeling (siGOLD) combined with serial-section transmission electron microscopy (ssTEM) to reveal the identity of multiple peptidergic neurons in a connectome. The unique antigenicity of neuropeptides, combined with their uniform distribution along axons, allowed us to identify distinct neurons by immunolabeling a small subset of sections within larger series. We demonstrate the scalability of siGOLD by identifying several peptidergic neurons using 11 neuropeptide antibodies on a full-body larval ssTEM dataset of the annelid Platynereis. We also reconstruct the circuitry of the sensory nuchal organs that were found by siGOLD to express the circadian neuropeptide pigment-dispersing factor. Our approach opens up the possibility for directly overlaying chemical neuromodulatory maps onto synaptic connectomic maps in the study of nervous systems.}, URL = {https://www.biorxiv.org/content/early/2015/09/23/027458}, eprint = {https://www.biorxiv.org/content/early/2015/09/23/027458.full.pdf}, journal = {bioRxiv} }