title: Golgi depletion from living cells with laser nanosurgery.
abstract: How Golgi biogenesis occurs in mammalian cells is a controversial problem. Can the Golgi complex (GC) form de novo from ER membranes or does it require a template? The method described in this chapter uses growth of cells on micropatterns to displace the GC from its juxtanuclear position and laser nanosurgery to subsequently deplete it from living cells. Golgi-depleted karyoplasts can be followed by time-lapse microscopy to address if and how the GC can be de novo synthesized from ER membranes. Furthermore, the study of different processes in the absence of the GC can shed light on the role of this organelle in the intracellular signaling and homeostasis.
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
authors.forename: Paolo
authors.initials: P
authors.lastname: Ronchi
authors.forename: Rainer
authors.initials: R
authors.lastname: Pepperkok
mesh_terms: D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002460:Cell Line; D002522:Chlorocebus aethiops; D005353:Fibronectins / Q000378:metabolism; D006056:Golgi Apparatus / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D061346:Laser Capture Microdissection; D008856:Microscopy, Fluorescence; D059008:Time-Lapse Imaging
chemical_list: D005353:Fibronectins; D049452:Green Fluorescent Proteins
keywords: Cell shaping; Golgi biogenesis; Laser nanosurgery; Micropattern; Subcellular dissection