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Nanoscaled RIM clustering at presynaptic active zones revealed by endogenous tagging
Chemical synaptic transmission involves neurotransmitter release from presynaptic active zones (AZs). The AZ protein Rab-3-interacting molecule (RIM) is important for normal Ca(2+)-triggered release. However, its precise localization within AZs of the glutamatergic neuromuscular junctions of Drosoph...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Life Science Alliance LLC
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10494931/ https://www.ncbi.nlm.nih.gov/pubmed/37696575 http://dx.doi.org/10.26508/lsa.202302021 |
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author | Mrestani, Achmed Dannhäuser, Sven Pauli, Martin Kollmannsberger, Philip Hübsch, Martha Morris, Lydia Langenhan, Tobias Heckmann, Manfred Paul, Mila M |
author_facet | Mrestani, Achmed Dannhäuser, Sven Pauli, Martin Kollmannsberger, Philip Hübsch, Martha Morris, Lydia Langenhan, Tobias Heckmann, Manfred Paul, Mila M |
author_sort | Mrestani, Achmed |
collection | PubMed |
description | Chemical synaptic transmission involves neurotransmitter release from presynaptic active zones (AZs). The AZ protein Rab-3-interacting molecule (RIM) is important for normal Ca(2+)-triggered release. However, its precise localization within AZs of the glutamatergic neuromuscular junctions of Drosophila melanogaster remains elusive. We used CRISPR/Cas9-assisted genome engineering of the rim locus to incorporate small epitope tags for targeted super-resolution imaging. A V5-tag, derived from simian virus 5, and an HA-tag, derived from human influenza virus, were N-terminally fused to the RIM Zinc finger. Whereas both variants are expressed in co-localization with the core AZ scaffold Bruchpilot, electrophysiological characterization reveals that AP-evoked synaptic release is disturbed in rim(V5−Znf) but not in rim(HA−Znf). In addition, rim(HA−Znf) synapses show intact presynaptic homeostatic potentiation. Combining super-resolution localization microscopy and hierarchical clustering, we detect ∼10 RIM(HA−Znf) subclusters with ∼13 nm diameter per AZ that are compacted and increased in numbers in presynaptic homeostatic potentiation. |
format | Online Article Text |
id | pubmed-10494931 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Life Science Alliance LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-104949312023-09-12 Nanoscaled RIM clustering at presynaptic active zones revealed by endogenous tagging Mrestani, Achmed Dannhäuser, Sven Pauli, Martin Kollmannsberger, Philip Hübsch, Martha Morris, Lydia Langenhan, Tobias Heckmann, Manfred Paul, Mila M Life Sci Alliance Research Articles Chemical synaptic transmission involves neurotransmitter release from presynaptic active zones (AZs). The AZ protein Rab-3-interacting molecule (RIM) is important for normal Ca(2+)-triggered release. However, its precise localization within AZs of the glutamatergic neuromuscular junctions of Drosophila melanogaster remains elusive. We used CRISPR/Cas9-assisted genome engineering of the rim locus to incorporate small epitope tags for targeted super-resolution imaging. A V5-tag, derived from simian virus 5, and an HA-tag, derived from human influenza virus, were N-terminally fused to the RIM Zinc finger. Whereas both variants are expressed in co-localization with the core AZ scaffold Bruchpilot, electrophysiological characterization reveals that AP-evoked synaptic release is disturbed in rim(V5−Znf) but not in rim(HA−Znf). In addition, rim(HA−Znf) synapses show intact presynaptic homeostatic potentiation. Combining super-resolution localization microscopy and hierarchical clustering, we detect ∼10 RIM(HA−Znf) subclusters with ∼13 nm diameter per AZ that are compacted and increased in numbers in presynaptic homeostatic potentiation. Life Science Alliance LLC 2023-09-11 /pmc/articles/PMC10494931/ /pubmed/37696575 http://dx.doi.org/10.26508/lsa.202302021 Text en © 2023 Mrestani et al. https://creativecommons.org/licenses/by/4.0/This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Articles Mrestani, Achmed Dannhäuser, Sven Pauli, Martin Kollmannsberger, Philip Hübsch, Martha Morris, Lydia Langenhan, Tobias Heckmann, Manfred Paul, Mila M Nanoscaled RIM clustering at presynaptic active zones revealed by endogenous tagging |
title | Nanoscaled RIM clustering at presynaptic active zones revealed by endogenous tagging |
title_full | Nanoscaled RIM clustering at presynaptic active zones revealed by endogenous tagging |
title_fullStr | Nanoscaled RIM clustering at presynaptic active zones revealed by endogenous tagging |
title_full_unstemmed | Nanoscaled RIM clustering at presynaptic active zones revealed by endogenous tagging |
title_short | Nanoscaled RIM clustering at presynaptic active zones revealed by endogenous tagging |
title_sort | nanoscaled rim clustering at presynaptic active zones revealed by endogenous tagging |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10494931/ https://www.ncbi.nlm.nih.gov/pubmed/37696575 http://dx.doi.org/10.26508/lsa.202302021 |
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