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...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Tina Ghelani, Carolina Montenegro-Venegas, Anna Fejtova, Thomas Dresbach
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	Golgi STED Bassoon synapse assembly synaptogenesis Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571
Acceso en línea:	https://doaj.org/article/293f35936d2f4672a52f113d0883329e
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:293f35936d2f4672a52f113d0883329e
record_format	dspace
spelling	oai:doaj.org-article:293f35936d2f4672a52f113d0883329e2021-11-11T10:39:04ZNanoscopical Analysis Reveals an Orderly Arrangement of the Presynaptic Scaffold Protein Bassoon at the Golgi-Apparatus1662-509910.3389/fnmol.2021.744034https://doaj.org/article/293f35936d2f4672a52f113d0883329e2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fnmol.2021.744034/fullhttps://doaj.org/toc/1662-5099Bassoon 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.Tina GhelaniCarolina Montenegro-VenegasCarolina Montenegro-VenegasCarolina Montenegro-VenegasAnna FejtovaAnna FejtovaAnna FejtovaThomas DresbachFrontiers Media S.A.articleGolgiSTEDBassoonsynapse assemblysynaptogenesisNeurosciences. Biological psychiatry. NeuropsychiatryRC321-571ENFrontiers in Molecular Neuroscience, Vol 14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Golgi STED Bassoon synapse assembly synaptogenesis Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571
spellingShingle	Golgi STED Bassoon synapse assembly synaptogenesis Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571 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
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.
format	article
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
title	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_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_sort	nanoscopical analysis reveals an orderly arrangement of the presynaptic scaffold protein bassoon at the golgi-apparatus
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/293f35936d2f4672a52f113d0883329e
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
_version_	1718439110321897472

Nanoscopical Analysis Reveals an Orderly Arrangement of the Presynaptic Scaffold Protein Bassoon at the Golgi-Apparatus

Ejemplares similares