TY - JOUR TI - A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes AU - Shahidi, Réza AU - Williams, Elizabeth A AU - Conzelmann, Markus AU - Asadulina, Albina AU - Verasztó, Csaba AU - Jasek, Sanja AU - Bezares-Calderón, Luis A AU - Jékely, Gáspár A2 - Calabrese, Ronald L VL - 4 PY - 2015 DA - 2015/12/15 SP - e11147 C1 - eLife 2015;4:e11147 DO - 10.7554/eLife.11147 UR - https://doi.org/10.7554/eLife.11147 AB - 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. KW - Platynereis dumerilii KW - annelid KW - neuropeptide KW - connectome KW - immunogold JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -