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

Content Provider	PubMed Central
Author	Shahidi, Réza Williams, Elizabeth A. Conzelmann, Markus Albina, Asadulina Csaba, Verasztó Jasek, Sanja Bezares-calderón, Luis A. Gáspár, Jékely
Editor	Calabrese, Ronald L.
Copyright Year	2015
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.
Related Links	http://dx.doi.org/10.7554/elife.11147
Starting Page	11147
File Format	PDF
ISSN	2050084X
e-ISSN	2050084X
Journal	eLife
Volume Number	4
Language	English
Publisher	eLife Sciences Publications, Ltd
Publisher Date	2015-12-01
Access Restriction	Open
Rights Holder	eLife Sciences Publications, Ltd
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Neuroscience Immunology and Microbiology Medicine Biochemistry, Genetics and Molecular Biology

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes

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

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