Vti1a/b support distinct aspects of TGN and cis-/medial Golgi organization

Danique M. van Bommel; Ruud F. Toonen; Matthijs Verhage

doi:10.1038/s41598-022-25331-x

Vti1a/b support distinct aspects of TGN and cis-/medial Golgi organization

Danique M. van Bommel, Ruud F. Toonen, Matthijs Verhage^*

^*Corresponding author for this work

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

Retrograde trafficking towards the trans-Golgi network (TGN) is important for dense core vesicle (DCV) biogenesis. Here, we used Vti1a/b deficient neurons to study the impact of disturbed retrograde trafficking on Golgi organization and cargo sorting. In Vti1a/b deficient neurons, staining intensity of cis-/medial Golgi proteins (e.g., GM130 and giantin) was increased, while the intensity of two recycling TGN proteins, TGN38 and TMEM87A, was decreased and the TGN-resident protein Golgin97 was normal. Levels and localization of DCV cargo markers, LAMP1 and KDEL were also altered. This phenotype was not caused by reduced Golgi size or absence of a TGN compartment. The phenotype was partially phenocopied by disturbing sphingolipid homeostasis, but was not rescued by overexpression of sphingomyelin synthases or the sphingolipid synthesis inhibitor myriocin. We conclude that Vti1a/b are important for distinct aspects of TGN and cis-/medial Golgi organization. Our data underline the importance of retrograde trafficking for Golgi organization, DCV cargo sorting and the distribution of proteins of the regulated secretory pathway.

Original language	English
Article number	20870
Pages (from-to)	1-12
Number of pages	12
Journal	Scientific Reports
Volume	12
DOIs	https://doi.org/10.1038/s41598-022-25331-x
Publication status	Published - 2 Dec 2022

Bibliographical note

Funding Information:
We thank Ingrid Saarloos and Robbert Zalm for cloning and producing viral particles; Lisa Laan, Ingrid Saarloos and Desiree Schut for producing glia and for primary culture assistance; Ingrid Saarloos for Western blotting; Joke Wortel for animal breeding; Joost Hoetjes for genotyping; and members of the Center for Neurogenomics and Cognitive Research DCV project team for fruitful discussions. We acknowledge the Microscopy and Cytometry Core Facility at the Amsterdam UMC—Location VUmc for providing assistance in microscopy. This work is supported by a European Research Council Advanced Grant (322966) of the European Union (to M.V.).

Funding Information:
We thank Ingrid Saarloos and Robbert Zalm for cloning and producing viral particles; Lisa Laan, Ingrid Saarloos and Desiree Schut for producing glia and for primary culture assistance; Ingrid Saarloos for Western blotting; Joke Wortel for animal breeding; Joost Hoetjes for genotyping; and members of the Center for Neurogenomics and Cognitive Research DCV project team for fruitful discussions. We acknowledge the Microscopy and Cytometry Core Facility at the Amsterdam UMC—Location VUmc for providing assistance in microscopy. This work is supported by a European Research Council Advanced Grant (322966) of the European Union (to M.V.).

Publisher Copyright:
© 2022, The Author(s).

Funding

We thank Ingrid Saarloos and Robbert Zalm for cloning and producing viral particles; Lisa Laan, Ingrid Saarloos and Desiree Schut for producing glia and for primary culture assistance; Ingrid Saarloos for Western blotting; Joke Wortel for animal breeding; Joost Hoetjes for genotyping; and members of the Center for Neurogenomics and Cognitive Research DCV project team for fruitful discussions. We acknowledge the Microscopy and Cytometry Core Facility at the Amsterdam UMC—Location VUmc for providing assistance in microscopy. This work is supported by a European Research Council Advanced Grant (322966) of the European Union (to M.V.). We thank Ingrid Saarloos and Robbert Zalm for cloning and producing viral particles; Lisa Laan, Ingrid Saarloos and Desiree Schut for producing glia and for primary culture assistance; Ingrid Saarloos for Western blotting; Joke Wortel for animal breeding; Joost Hoetjes for genotyping; and members of the Center for Neurogenomics and Cognitive Research DCV project team for fruitful discussions. We acknowledge the Microscopy and Cytometry Core Facility at the Amsterdam UMC—Location VUmc for providing assistance in microscopy. This work is supported by a European Research Council Advanced Grant (322966) of the European Union (to M.V.).

Funders	Funder number
European Research Council
European Commission
Center for Neurogenomics and Cognitive Research DCV
Seventh Framework Programme	322966

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title = "Vti1a/b support distinct aspects of TGN and cis-/medial Golgi organization",

abstract = "Retrograde trafficking towards the trans-Golgi network (TGN) is important for dense core vesicle (DCV) biogenesis. Here, we used Vti1a/b deficient neurons to study the impact of disturbed retrograde trafficking on Golgi organization and cargo sorting. In Vti1a/b deficient neurons, staining intensity of cis-/medial Golgi proteins (e.g., GM130 and giantin) was increased, while the intensity of two recycling TGN proteins, TGN38 and TMEM87A, was decreased and the TGN-resident protein Golgin97 was normal. Levels and localization of DCV cargo markers, LAMP1 and KDEL were also altered. This phenotype was not caused by reduced Golgi size or absence of a TGN compartment. The phenotype was partially phenocopied by disturbing sphingolipid homeostasis, but was not rescued by overexpression of sphingomyelin synthases or the sphingolipid synthesis inhibitor myriocin. We conclude that Vti1a/b are important for distinct aspects of TGN and cis-/medial Golgi organization. Our data underline the importance of retrograde trafficking for Golgi organization, DCV cargo sorting and the distribution of proteins of the regulated secretory pathway.",

author = "{van Bommel}, {Danique M.} and Toonen, {Ruud F.} and Matthijs Verhage",

note = "Funding Information: We thank Ingrid Saarloos and Robbert Zalm for cloning and producing viral particles; Lisa Laan, Ingrid Saarloos and Desiree Schut for producing glia and for primary culture assistance; Ingrid Saarloos for Western blotting; Joke Wortel for animal breeding; Joost Hoetjes for genotyping; and members of the Center for Neurogenomics and Cognitive Research DCV project team for fruitful discussions. We acknowledge the Microscopy and Cytometry Core Facility at the Amsterdam UMC—Location VUmc for providing assistance in microscopy. This work is supported by a European Research Council Advanced Grant (322966) of the European Union (to M.V.). Funding Information: We thank Ingrid Saarloos and Robbert Zalm for cloning and producing viral particles; Lisa Laan, Ingrid Saarloos and Desiree Schut for producing glia and for primary culture assistance; Ingrid Saarloos for Western blotting; Joke Wortel for animal breeding; Joost Hoetjes for genotyping; and members of the Center for Neurogenomics and Cognitive Research DCV project team for fruitful discussions. We acknowledge the Microscopy and Cytometry Core Facility at the Amsterdam UMC—Location VUmc for providing assistance in microscopy. This work is supported by a European Research Council Advanced Grant (322966) of the European Union (to M.V.). Publisher Copyright: {\textcopyright} 2022, The Author(s).",

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doi = "10.1038/s41598-022-25331-x",

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AU - Toonen, Ruud F.

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N1 - Funding Information: We thank Ingrid Saarloos and Robbert Zalm for cloning and producing viral particles; Lisa Laan, Ingrid Saarloos and Desiree Schut for producing glia and for primary culture assistance; Ingrid Saarloos for Western blotting; Joke Wortel for animal breeding; Joost Hoetjes for genotyping; and members of the Center for Neurogenomics and Cognitive Research DCV project team for fruitful discussions. We acknowledge the Microscopy and Cytometry Core Facility at the Amsterdam UMC—Location VUmc for providing assistance in microscopy. This work is supported by a European Research Council Advanced Grant (322966) of the European Union (to M.V.). Funding Information: We thank Ingrid Saarloos and Robbert Zalm for cloning and producing viral particles; Lisa Laan, Ingrid Saarloos and Desiree Schut for producing glia and for primary culture assistance; Ingrid Saarloos for Western blotting; Joke Wortel for animal breeding; Joost Hoetjes for genotyping; and members of the Center for Neurogenomics and Cognitive Research DCV project team for fruitful discussions. We acknowledge the Microscopy and Cytometry Core Facility at the Amsterdam UMC—Location VUmc for providing assistance in microscopy. This work is supported by a European Research Council Advanced Grant (322966) of the European Union (to M.V.). Publisher Copyright: © 2022, The Author(s).

PY - 2022/12/2

Y1 - 2022/12/2

N2 - Retrograde trafficking towards the trans-Golgi network (TGN) is important for dense core vesicle (DCV) biogenesis. Here, we used Vti1a/b deficient neurons to study the impact of disturbed retrograde trafficking on Golgi organization and cargo sorting. In Vti1a/b deficient neurons, staining intensity of cis-/medial Golgi proteins (e.g., GM130 and giantin) was increased, while the intensity of two recycling TGN proteins, TGN38 and TMEM87A, was decreased and the TGN-resident protein Golgin97 was normal. Levels and localization of DCV cargo markers, LAMP1 and KDEL were also altered. This phenotype was not caused by reduced Golgi size or absence of a TGN compartment. The phenotype was partially phenocopied by disturbing sphingolipid homeostasis, but was not rescued by overexpression of sphingomyelin synthases or the sphingolipid synthesis inhibitor myriocin. We conclude that Vti1a/b are important for distinct aspects of TGN and cis-/medial Golgi organization. Our data underline the importance of retrograde trafficking for Golgi organization, DCV cargo sorting and the distribution of proteins of the regulated secretory pathway.

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Vti1a/b support distinct aspects of TGN and cis-/medial Golgi organization

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