Nanoscopical Analysis Reveals an Orderly Arrangement of the Presynaptic Scaffold Protein Bassoon at the Golgi-Apparatus
Bassoon is a core scaffold protein of the presynaptic active zone. In brain synapses, the C-terminus of Bassoon is oriented toward the plasma membrane and its N-terminus is oriented toward synaptic vesicles. At the Golgi-apparatus, Bassoon is thought to assemble active zone precursor structures, but...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2021-11-01
|
| Series: | Frontiers in Molecular Neuroscience |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fnmol.2021.744034/full |
| _version_ | 1818821153387446272 |
|---|---|
| author | Tina Ghelani Carolina Montenegro-Venegas Carolina Montenegro-Venegas Carolina Montenegro-Venegas Anna Fejtova Anna Fejtova Anna Fejtova Thomas Dresbach |
| author_facet | Tina Ghelani Carolina Montenegro-Venegas Carolina Montenegro-Venegas Carolina Montenegro-Venegas Anna Fejtova Anna Fejtova Anna Fejtova Thomas Dresbach |
| author_sort | Tina Ghelani |
| collection | DOAJ |
| description | Bassoon is a core scaffold protein of the presynaptic active zone. In brain synapses, the C-terminus of Bassoon is oriented toward the plasma membrane and its N-terminus is oriented toward synaptic vesicles. At the Golgi-apparatus, Bassoon is thought to assemble active zone precursor structures, but whether it is arranged in an orderly fashion is unknown. Understanding the topology of this large scaffold protein is important for models of active zone biogenesis. Using stimulated emission depletion nanoscopy in cultured hippocampal neurons, we found that an N-terminal intramolecular tag of recombinant Bassoon, but not C-terminal tag, colocalized with markers of the trans-Golgi network (TGN). The N-terminus of Bassoon was located between 48 and 69 nm away from TGN38, while its C-terminus was located between 100 and 115 nm away from TGN38. Sequences within the first 95 amino acids of Bassoon were required for this arrangement. Our results indicate that, at the Golgi-apparatus, Bassoon is oriented with its N-terminus toward and its C-terminus away from the trans Golgi network membrane. Moreover, they suggest that Bassoon is an extended molecule at the trans Golgi network with the distance between amino acids 97 and 3,938, estimated to be between 46 and 52 nm. Our data are consistent with a model, in which the N-terminus of Bassoon binds to the membranes of the trans-Golgi network, while the C-terminus associates with active zone components, thus reflecting the topographic arrangement characteristic of synapses also at the Golgi-apparatus. |
| first_indexed | 2024-12-18T23:03:40Z |
| format | Article |
| id | doaj.art-293f35936d2f4672a52f113d0883329e |
| institution | Directory Open Access Journal |
| issn | 1662-5099 |
| language | English |
| last_indexed | 2024-12-18T23:03:40Z |
| publishDate | 2021-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Neuroscience |
| spelling | doaj.art-293f35936d2f4672a52f113d0883329e2022-12-21T20:48:31ZengFrontiers Media S.A.Frontiers in Molecular Neuroscience1662-50992021-11-011410.3389/fnmol.2021.744034744034Nanoscopical Analysis Reveals an Orderly Arrangement of the Presynaptic Scaffold Protein Bassoon at the Golgi-ApparatusTina Ghelani0Carolina Montenegro-Venegas1Carolina Montenegro-Venegas2Carolina Montenegro-Venegas3Anna Fejtova4Anna Fejtova5Anna Fejtova6Thomas Dresbach7Institute of Anatomy and Embryology, University Medical Center Göttingen, Göttingen, GermanyDepartment of Neurochemistry and Molecular Biology, Leibniz Institute for Neurobiology, Magdeburg, GermanyCenter for Behavioral Brain Sciences, Magdeburg, GermanyInstitute for Pharmacology and Toxicology, Medical Faculty, Otto von Guericke University, Magdeburg, GermanyDepartment of Neurochemistry and Molecular Biology, Leibniz Institute for Neurobiology, Magdeburg, GermanyDepartment of Psychiatry and Psychotherapy, University Hospital Erlangen, Friedrich-Alexander Universität Erlangen-Nürnberg, Erlangen, GermanyRG Presynaptic Plasticity, Leibniz Institute for Neurobiology, Magdeburg, GermanyInstitute of Anatomy and Embryology, University Medical Center Göttingen, Göttingen, GermanyBassoon is a core scaffold protein of the presynaptic active zone. In brain synapses, the C-terminus of Bassoon is oriented toward the plasma membrane and its N-terminus is oriented toward synaptic vesicles. At the Golgi-apparatus, Bassoon is thought to assemble active zone precursor structures, but whether it is arranged in an orderly fashion is unknown. Understanding the topology of this large scaffold protein is important for models of active zone biogenesis. Using stimulated emission depletion nanoscopy in cultured hippocampal neurons, we found that an N-terminal intramolecular tag of recombinant Bassoon, but not C-terminal tag, colocalized with markers of the trans-Golgi network (TGN). The N-terminus of Bassoon was located between 48 and 69 nm away from TGN38, while its C-terminus was located between 100 and 115 nm away from TGN38. Sequences within the first 95 amino acids of Bassoon were required for this arrangement. Our results indicate that, at the Golgi-apparatus, Bassoon is oriented with its N-terminus toward and its C-terminus away from the trans Golgi network membrane. Moreover, they suggest that Bassoon is an extended molecule at the trans Golgi network with the distance between amino acids 97 and 3,938, estimated to be between 46 and 52 nm. Our data are consistent with a model, in which the N-terminus of Bassoon binds to the membranes of the trans-Golgi network, while the C-terminus associates with active zone components, thus reflecting the topographic arrangement characteristic of synapses also at the Golgi-apparatus.https://www.frontiersin.org/articles/10.3389/fnmol.2021.744034/fullGolgiSTEDBassoonsynapse assemblysynaptogenesis |
| spellingShingle | Tina Ghelani Carolina Montenegro-Venegas Carolina Montenegro-Venegas Carolina Montenegro-Venegas Anna Fejtova Anna Fejtova Anna Fejtova Thomas Dresbach Nanoscopical Analysis Reveals an Orderly Arrangement of the Presynaptic Scaffold Protein Bassoon at the Golgi-Apparatus Frontiers in Molecular Neuroscience Golgi STED Bassoon synapse assembly synaptogenesis |
| title | Nanoscopical Analysis Reveals an Orderly Arrangement of the Presynaptic Scaffold Protein Bassoon at the Golgi-Apparatus |
| title_full | Nanoscopical Analysis Reveals an Orderly Arrangement of the Presynaptic Scaffold Protein Bassoon at the Golgi-Apparatus |
| title_fullStr | Nanoscopical Analysis Reveals an Orderly Arrangement of the Presynaptic Scaffold Protein Bassoon at the Golgi-Apparatus |
| title_full_unstemmed | Nanoscopical Analysis Reveals an Orderly Arrangement of the Presynaptic Scaffold Protein Bassoon at the Golgi-Apparatus |
| title_short | Nanoscopical Analysis Reveals an Orderly Arrangement of the Presynaptic Scaffold Protein Bassoon at the Golgi-Apparatus |
| title_sort | nanoscopical analysis reveals an orderly arrangement of the presynaptic scaffold protein bassoon at the golgi apparatus |
| topic | Golgi STED Bassoon synapse assembly synaptogenesis |
| url | https://www.frontiersin.org/articles/10.3389/fnmol.2021.744034/full |
| work_keys_str_mv | AT tinaghelani nanoscopicalanalysisrevealsanorderlyarrangementofthepresynapticscaffoldproteinbassoonatthegolgiapparatus AT carolinamontenegrovenegas nanoscopicalanalysisrevealsanorderlyarrangementofthepresynapticscaffoldproteinbassoonatthegolgiapparatus AT carolinamontenegrovenegas nanoscopicalanalysisrevealsanorderlyarrangementofthepresynapticscaffoldproteinbassoonatthegolgiapparatus AT carolinamontenegrovenegas nanoscopicalanalysisrevealsanorderlyarrangementofthepresynapticscaffoldproteinbassoonatthegolgiapparatus AT annafejtova nanoscopicalanalysisrevealsanorderlyarrangementofthepresynapticscaffoldproteinbassoonatthegolgiapparatus AT annafejtova nanoscopicalanalysisrevealsanorderlyarrangementofthepresynapticscaffoldproteinbassoonatthegolgiapparatus AT annafejtova nanoscopicalanalysisrevealsanorderlyarrangementofthepresynapticscaffoldproteinbassoonatthegolgiapparatus AT thomasdresbach nanoscopicalanalysisrevealsanorderlyarrangementofthepresynapticscaffoldproteinbassoonatthegolgiapparatus |