Golgi depletion from living cells with laser nanosurgery.

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.