Correlative Light-Electron Microscopy Reveals the Tubular-Saccular Ultrastructure of Carriers Operating between Golgi Apparatus and Plasma Membrane

doi:10.1083/jcb.148.1.45

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© The Rockefeller University Press, 0021-9525/2000/1/45/ $5.00
The Journal of Cell Biology, Volume 148, Number 1, January 10, 2000 45-58

Original Article

Correlative Light-Electron Microscopy Reveals the Tubular-Saccular Ultrastructure of Carriers Operating between Golgi Apparatus and Plasma Membrane

Roman S. Polishchuk^a, Elena V. Polishchuk^a, Pierfrancesco Marra^a, Saverio Alberti^a, Roberto Buccione^a, Alberto Luini^a, and Alexander A. Mironov^a
^a Department of Cell Biology and Oncology, Istituto di Ricerche Farmacologiche "Mario Negri," Consorzio Mario Negri Sud, 66030 S. Maria Imbaro (Chieti), Italy

Correspondence to: Alexander A. Mironov, Department of Cell Biology and Oncology, Consorzio Mario Negri Sud, Via Nazionale, 66030 S. Maria Imbaro (Chieti), Italy. Tel:39-0872-570-355 Fax:39-0872-570412 E-mail:mironov{at}cmns.mnegri.it.

Transport intermediates (TIs) have a central role in intracellulartraffic, and much effort has been directed towards definingtheir molecular organization. Unfortunately, major uncertaintiesremain regarding their true structure in living cells. To addressthis question, we have developed an approach based on the combinationof the green fluorescent protein technology and correlativelight-electron microscopy, by which it is possible to monitoran individual carrier in vivo and then take a picture of itsultrastructure at any moment of its lifecycle. We have appliedthis technique to define the structure of TIs operating fromthe Golgi apparatus to the plasma membrane, whose in vivo dynamicshave been characterized recently by light microscopy. We findthat these carriers are large (ranging from 0.3–1.7 µmin maximum diameter, nearly half the size of a Golgi cisterna),comprise almost exclusively tubular-saccular structures, andfuse directly with the plasma membrane, sometimes minutes afterdocking to the fusion site.

Key Words: Golgi complex, anterograde intracellular transport, membranetraffic, immunoelectron microscopy, plasmalemma

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