RT Journal Article
SR Electronic
T1 siGOLD: a Serial Multiplex Immunogold Labeling Method for Identifying Peptidergic Neurons in Connectomes
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 027458
DO 10.1101/027458
A1 Réza Shahidi
A1 Elizabeth Williams
A1 Markus Conzelmann
A1 Albina Asadulina
A1 Csaba Verasztó
A1 Sanja Jasek
A1 Luis A. Bezares-Calderón
A1 Gáspár Jékely
YR 2015
UL http://biorxiv.org/content/early/2015/09/23/027458.abstract
AB 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.