TY - JOUR
T1 - Vti1a/b support distinct aspects of TGN and cis-/medial Golgi organization
AU - van Bommel, Danique M.
AU - Toonen, Ruud F.
AU - Verhage, Matthijs
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 - 2022/12/1
Y1 - 2022/12/1
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.
AB - 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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U2 - https://doi.org/10.1038/s41598-022-25331-x
DO - https://doi.org/10.1038/s41598-022-25331-x
M3 - Article
C2 - 36460703
SN - 2045-2322
VL - 12
SP - 1
EP - 12
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 20870
ER -