Imaging Secretory Granule Budding from the Trans-Golgi Network Using Retention Using Selective Hook (RUSH)

Cedric S Asensio

doi:10.1007/978-1-0716-2209-4_3

Imaging Secretory Granule Budding from the Trans-Golgi Network Using Retention Using Selective Hook (RUSH)

Methods Mol Biol. 2022:2473:23-28. doi: 10.1007/978-1-0716-2209-4_3.

Author

Cedric S Asensio¹

Affiliation

¹ Department of Biological Sciences, University of Denver, Denver, CO, USA. Cedric.Asensio@du.edu.

PMID: 35819756
DOI: 10.1007/978-1-0716-2209-4_3

Abstract

The retention using selective hook (RUSH) system enables us to synchronize and visualize the movement of cargoes along the secretory pathway. A fluorescently tagged cargo of interest is retained in the endoplasmic reticulum and released in a biotin-dependent manner. Here, we report a detailed protocol describing the steps necessary to perform RUSH experiments to study secretory granule biogenesis at the trans-Golgi network (TGN).

Keywords: Cell biology; Golgi apparatus; Imaging; Insulin; Membrane trafficking; RUSH; Secretory granules; Trans-Golgi network; Vesicle budding.

MeSH terms

Endoplasmic Reticulum / metabolism
Golgi Apparatus* / metabolism
Secretory Pathway
Secretory Vesicles / metabolism
trans-Golgi Network* / metabolism

Grants and funding

R01 GM124035/GM/NIGMS NIH HHS/United States