A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes
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 transm...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2015-12-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/11147 |
| _version_ | 1828375471087157248 |
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| author | Réza Shahidi Elizabeth A Williams Markus Conzelmann Albina Asadulina Csaba Verasztó Sanja Jasek Luis A Bezares-Calderón Gáspár Jékely |
| author_facet | Réza Shahidi Elizabeth A Williams Markus Conzelmann Albina Asadulina Csaba Verasztó Sanja Jasek Luis A Bezares-Calderón Gáspár Jékely |
| author_sort | Réza Shahidi |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-14T07:47:03Z |
| format | Article |
| id | doaj.art-5440e12252ca4e9985d6dde42ebe34a6 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-14T07:47:03Z |
| publishDate | 2015-12-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-5440e12252ca4e9985d6dde42ebe34a62022-12-22T02:05:18ZengeLife Sciences Publications LtdeLife2050-084X2015-12-01410.7554/eLife.11147A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomesRéza Shahidi0Elizabeth A Williams1Markus Conzelmann2Albina Asadulina3Csaba Verasztó4Sanja Jasek5Luis A Bezares-Calderón6Gáspár Jékely7https://orcid.org/0000-0001-8496-9836Max-Planck-Institute for Developmental Biology, Tübingen, GermanyMax-Planck-Institute for Developmental Biology, Tübingen, GermanyMax-Planck-Institute for Developmental Biology, Tübingen, GermanyMax-Planck-Institute for Developmental Biology, Tübingen, GermanyMax-Planck-Institute for Developmental Biology, Tübingen, GermanyMax-Planck-Institute for Developmental Biology, Tübingen, GermanyMax-Planck-Institute for Developmental Biology, Tübingen, GermanyMax-Planck-Institute for Developmental Biology, Tübingen, GermanyElectron 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.https://elifesciences.org/articles/11147Platynereis dumeriliiannelidneuropeptideconnectomeimmunogold |
| spellingShingle | Réza Shahidi Elizabeth A Williams Markus Conzelmann Albina Asadulina Csaba Verasztó Sanja Jasek Luis A Bezares-Calderón Gáspár Jékely A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes eLife Platynereis dumerilii annelid neuropeptide connectome immunogold |
| title | A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes |
| title_full | A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes |
| title_fullStr | A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes |
| title_full_unstemmed | A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes |
| title_short | A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes |
| title_sort | serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes |
| topic | Platynereis dumerilii annelid neuropeptide connectome immunogold |
| url | https://elifesciences.org/articles/11147 |
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