Elucidation of intracellular recycling pathways leading to exocytosis of the Fc receptor, FcRn, by using multifocal plane microscopy.
The intracellular events on the recycling pathway that lead from sorting endosomes to exocytosis at the plasma membrane are central to cellular function. However, despite intensive study, these processes are poorly characterized in spatial and dynamic terms. The primary reason for this is that, to date, it has not been possible to visualize rapidly moving intracellular compartments in three dimensions in cells. Here, we use a recently developed imaging setup in which multiple planes can be simultaneously imaged within the cell in conjunction with visualization of the plasma membrane plane by using total internal reflection fluorescence microscopy. This has allowed us to track and characterize intracellular events on the recycling pathway that lead to exocytosis of the MHC Class I-related receptor, FcRn. We observe both direct delivery of tubular and vesicular transport containers (TCs) from sorting endosomes to exocytic sites at the plasma membrane, and indirect pathways in which TCs that are not in proximity to sorting endosomes undergo exocytosis. TCs can also interact with different sorting endosomes before exocytosis. Our data provide insight into the intracellular events that precede exocytic fusion.
Prabhat
Prashant
P
Department of Immunology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
Gan
Zhuo
Z
Chao
Jerry
J
Ram
Sripad
S
Vaccaro
Carlos
C
Gibbons
Steven
S
Ober
Raimund J
RJ
Ward
E Sally
ES
D018929:Cell Culture Techniques; D002460:Cell Line; D011992:Endosomes / Q000276:immunology*; D042783:Endothelial Cells / Q000276:immunology / Q000378:metabolism*; D005089:Exocytosis / Q000276:immunology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000235:genetics / Q000276:immunology*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D010957:Plasmids; D011961:Receptors, Fc / Q000235:genetics / Q000276:immunology*; D014162:Transfection; D001613:beta 2-Microglobulin / Q000378:metabolism
D015395:Histocompatibility Antigens Class I; D011961:Receptors, Fc; D001613:beta 2-Microglobulin; D049452:Green Fluorescent Proteins; C100688:Fc receptor, neonatal