DOI: 10.1242/jcs.00289. Real-time analysis of clathrin-mediated endocytosis during cell migration

Real-time analysis of clathrin-mediated endocytosis during cell migration

10.1242/jcs.00289

Crossref journal-article

The Company of Biologists

Journal of Cell Science (237)

Abstract

Simultaneous dual-color total-internal-reflection fluorescence microscopy(TIR-FM) was performed to analyze the internalization and distribution of markers for clathrin-mediated endocytosis (clathrin, dynamin1, dynamin2 and transferrin) in migrating cells. In MDCK cells, which endogenously express dynamin2, the dynamin2-EGFP fluorescence demonstrated identical spatial and temporal behavior as clathrin both prior to and during internalization. By contrast, in the same cells, the neuronal dynamin1 only localized with clathrin just prior to endocytosis. In migrating cells, each endocytic marker was polarized towards the leading edge, away from the lagging edge. These observations suggest a re-evaluation of the functional differences between dynamin1 and dynamin2, and of the role of clathrin-mediated endocytosis in cell migration.

Bibliography

Rappoport, J. Z., & Simon, S. M. (2003). Real-time analysis of clathrin-mediated endocytosis during cell migration. Journal of Cell Science, 116(5), 847â855.

Authors 2

Joshua Z. Rappoport (first)
Sanford M. Simon (additional)

References 46 Referenced 155

Dates

Type	When
Created	22 years, 6 months ago (Feb. 5, 2003, 9:02 p.m.)
Deposited	4 years, 4 months ago (April 25, 2021, 2:10 p.m.)
Indexed	2 months, 3 weeks ago (June 10, 2025, 5:12 a.m.)
Issued	22 years, 6 months ago (March 1, 2003)
Published	22 years, 6 months ago (March 1, 2003)
Published Print	22 years, 6 months ago (March 1, 2003)

Funders 0

None

BibTeX

@article{Rappoport_2003, title={Real-time analysis of clathrin-mediated endocytosis during cell migration}, volume={116}, ISSN={0021-9533}, url={http://dx.doi.org/10.1242/jcs.00289}, DOI={10.1242/jcs.00289}, number={5}, journal={Journal of Cell Science}, publisher={The Company of Biologists}, author={Rappoport, Joshua Z. and Simon, Sanford M.}, year={2003}, month=mar, pages={847–855} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        6,
        10
      ]
    ],
    "date-time": "2025-06-10T09:12:48Z",
    "timestamp": 1749546768400
  },
  "reference-count": 46,
  "publisher": "The Company of Biologists",
  "issue": "5",
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        2003,
        3,
        1
      ]
    ]
  },
  "abstract": "<jats:p>Simultaneous dual-color total-internal-reflection fluorescence microscopy(TIR-FM) was performed to analyze the internalization and distribution of markers for clathrin-mediated endocytosis (clathrin, dynamin1, dynamin2 and transferrin) in migrating cells. In MDCK cells, which endogenously express dynamin2, the dynamin2-EGFP fluorescence demonstrated identical spatial and temporal behavior as clathrin both prior to and during internalization. By contrast, in the same cells, the neuronal dynamin1 only localized with clathrin just prior to endocytosis. In migrating cells, each endocytic marker was polarized towards the leading edge, away from the lagging edge. These observations suggest a re-evaluation of the functional differences between dynamin1 and dynamin2, and of the role of clathrin-mediated endocytosis in cell migration.</jats:p>",
  "DOI": "10.1242/jcs.00289",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2003,
        2,
        6
      ]
    ],
    "date-time": "2003-02-06T02:02:33Z",
    "timestamp": 1044496953000
  },
  "page": "847-855",
  "source": "Crossref",
  "is-referenced-by-count": 155,
  "title": "Real-time analysis of clathrin-mediated endocytosis during cell migration",
  "prefix": "10.1242",
  "volume": "116",
  "author": [
    {
      "given": "Joshua Z.",
      "family": "Rappoport",
      "sequence": "first",
      "affiliation": [
        {
          "name": "The Laboratory of Cellular Biophysics, The Rockefeller University, 1230 York Avenue, Box 304, New York, NY 10021, USA"
        }
      ]
    },
    {
      "given": "Sanford M.",
      "family": "Simon",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "The Laboratory of Cellular Biophysics, The Rockefeller University, 1230 York Avenue, Box 304, New York, NY 10021, USA"
        }
      ]
    }
  ],
  "member": "237",
  "reference": [
    {
      "key": "2021042517532726100_REF1",
      "doi-asserted-by": "crossref",
      "unstructured": "Altschuler, Y., Barbas, S. M., Terlecky, L. J., Tang, K., Hardy,S., Mostov, K. E. and Schmid, S. L. (1998). Redundant and distinct functions for dynamin-1 and dynamin-2 isoforms. J. Cell Biol.143,1871-1881.",
      "DOI": "10.1083/jcb.143.7.1871"
    },
    {
      "key": "2021042517532726100_REF2",
      "doi-asserted-by": "crossref",
      "unstructured": "Apodaca, G. (2001). Endocytic traffic in polarized epithelial cells: role of the actin and microtubule cytoskeleton. Traffic2,149-159.",
      "DOI": "10.1034/j.1600-0854.2001.020301.x"
    },
    {
      "key": "2021042517532726100_REF3",
      "doi-asserted-by": "crossref",
      "unstructured": "Axelrod, D. (1981). Cell-substrate contacts illuminated by total internal reflection fluorescence. J. Cell Biol.89,141-145.",
      "DOI": "10.1083/jcb.89.1.141"
    },
    {
      "key": "2021042517532726100_REF4",
      "doi-asserted-by": "crossref",
      "unstructured": "Bajno, L., Peng, X. R., Schreiber, A. D., Moore, H. P., Trimble,W. S. and Grinstein, S. (2000). Focal exocytosis of VAMP3-containing vesicles at sites of phagosome formation. J. Cell Biol.149,697-706.",
      "DOI": "10.1083/jcb.149.3.697"
    },
    {
      "key": "2021042517532726100_REF5",
      "doi-asserted-by": "crossref",
      "unstructured": "Benmerah, A., Bayrou, M., Cerf-Bensussan, N. and Dautry-Varsat,A. (1999). Inhibition of clathrin-coated pit assembly by an Eps15 mutant. J. Cell Sci.112,1303-1311.",
      "DOI": "10.1242/jcs.112.9.1303"
    },
    {
      "key": "2021042517532726100_REF6",
      "doi-asserted-by": "crossref",
      "unstructured": "Benmerah, A., Lamaze, C., Begue, B., Schmid, S. L.,Dautry-Varsat, A. and Cerf-Bensussan, N. (1998). AP-2/Eps15 interaction is required for receptor-mediated endocytosis. J. Cell Biol.140,1055-1062.",
      "DOI": "10.1083/jcb.140.5.1055"
    },
    {
      "key": "2021042517532726100_REF7",
      "doi-asserted-by": "crossref",
      "unstructured": "Bretscher, M. S. (1996). Moving membrane up to the front of migrating cells. Cell85,465-467.",
      "DOI": "10.1016/S0092-8674(00)81246-5"
    },
    {
      "key": "2021042517532726100_REF8",
      "doi-asserted-by": "crossref",
      "unstructured": "Cao, H., Garcia, F. and McNiven, M. A. (1998). Differential distribution of dynamin isoforms in mammalian cells. Mol. Biol. Cell9,2595-2609.",
      "DOI": "10.1091/mbc.9.9.2595"
    },
    {
      "key": "2021042517532726100_REF9",
      "doi-asserted-by": "crossref",
      "unstructured": "Carter, R. E. and Sorkin, A. (1998). Endocytosis of functional epidermal growth factor receptor-green fluorescent protein chimera. J. Biol. Chem.273,35000-35007.",
      "DOI": "10.1074/jbc.273.52.35000"
    },
    {
      "key": "2021042517532726100_REF10",
      "doi-asserted-by": "crossref",
      "unstructured": "Collins, B. M., McCoy, A. J., Kent, H. M., Evans, P. R. and Owen, D. J. (2002). Molecular architecture and functional model of the endocytic AP2 complex. Cell109,523-535.",
      "DOI": "10.1016/S0092-8674(02)00735-3"
    },
    {
      "key": "2021042517532726100_REF11",
      "doi-asserted-by": "crossref",
      "unstructured": "Conrad, M. E., Umbreit, J. N. and Moore, E. G.(1999). Iron absorption and transport. Am. J. Med. Sci.318,213-229.",
      "DOI": "10.1016/S0002-9629(15)40626-3"
    },
    {
      "key": "2021042517532726100_REF12",
      "doi-asserted-by": "crossref",
      "unstructured": "de Curtis, I (2001). Cell migration: GAPs between membrane traffic and the cytoskeleton. EMBO Rep.2,277-281.",
      "DOI": "10.1093/embo-reports/kve072"
    },
    {
      "key": "2021042517532726100_REF13",
      "doi-asserted-by": "crossref",
      "unstructured": "Doxsey, S. J., Brodsky, F. M., Blank, G. S. and Helenius, A.(1987). Inhibition of endocytosis by anti-clathrin antibodies. Cell50,453-463.",
      "DOI": "10.1016/0092-8674(87)90499-5"
    },
    {
      "key": "2021042517532726100_REF14",
      "doi-asserted-by": "crossref",
      "unstructured": "Engqvist-Goldstein, A. E., Warren, R. A., Kessels, M. M., Keen,J. H., Heuser, J. and Drubin, D. G. (2001). The actin-binding protein Hip1R associates with clathrin during early stages of endocytosis and promotes clathrin assembly in vitro. J. Cell Biol.154,1209-1223.",
      "DOI": "10.1083/jcb.200106089"
    },
    {
      "key": "2021042517532726100_REF15",
      "doi-asserted-by": "crossref",
      "unstructured": "Fenteany, G., Janmey, P. A. and Stossel, T. P.(2000). Signaling pathways and cell mechanics involved in wound closure by epithelial cell sheets. Curr. Biol.10,831-838.",
      "DOI": "10.1016/S0960-9822(00)00579-0"
    },
    {
      "key": "2021042517532726100_REF16",
      "doi-asserted-by": "crossref",
      "unstructured": "Fialka, I., Steinlein, P., Ahorn, H., Bock, G., Burbelo, P. D.,Haberfellner, M., Lottspeich, F., Paiha, K., Pasquali, C. and Huber, L. A.(1999). Identification of syntenin as a protein of the apical early endocytic compartment in Madin-Darby canine kidney cells. J. Biol. Chem.274,26233-26239.",
      "DOI": "10.1074/jbc.274.37.26233"
    },
    {
      "key": "2021042517532726100_REF17",
      "doi-asserted-by": "crossref",
      "unstructured": "Fuller, S. D. and Simons, K. (1986). Transferrin receptor polarity and recycling accuracy in `tight' and `leaky'strains of Madin-Darby canine kidney cells. J. Cell Biol.103,1767-1779.",
      "DOI": "10.1083/jcb.103.5.1767"
    },
    {
      "key": "2021042517532726100_REF18",
      "doi-asserted-by": "crossref",
      "unstructured": "Gaidarov, I., Santini, F., Warren, R. A. and Keen, J. H.(1999). Spatial control of coated-pit dynamics in living cells. Nat. Cell Biol.1,1-7.",
      "DOI": "10.1038/8971"
    },
    {
      "key": "2021042517532726100_REF19",
      "doi-asserted-by": "crossref",
      "unstructured": "Goldberg, M. B. (2001). Actin-based motility of intracellular microbial pathogens. Microbiol. Mol. Biol. Rev.65,595-626.",
      "DOI": "10.1128/MMBR.65.4.595-626.2001"
    },
    {
      "key": "2021042517532726100_REF20",
      "doi-asserted-by": "crossref",
      "unstructured": "Hopkins, C. R., Miller, K. and Beardmore, J. M.(1985). Receptor-mediated endocytosis of transferrin and epidermal growth factor receptors: a comparison of constitutive and ligand-induced uptake. J. Cell Sci.3 (Suppl.),173-186.",
      "DOI": "10.1242/jcs.1985.Supplement_3.17"
    },
    {
      "key": "2021042517532726100_REF21",
      "doi-asserted-by": "crossref",
      "unstructured": "Jiang, W. and Hunter, T. (1998). Analysis of cell-cycle profiles in transfected cells using a membrane- targeted GFP. Biotechniques24,349, 350, 352, 354.",
      "DOI": "10.2144/98243bm03"
    },
    {
      "key": "2021042517532726100_REF22",
      "doi-asserted-by": "crossref",
      "unstructured": "Kamiguchi, H. and Lemmon, V. (2000). Recycling of the cell adhesion molecule L1 in axonal growth cones. J. Neurosci.20,3676-3686.",
      "DOI": "10.1523/JNEUROSCI.20-10-03676.2000"
    },
    {
      "key": "2021042517532726100_REF23",
      "doi-asserted-by": "crossref",
      "unstructured": "Kamiguchi, H., Long, K. E., Pendergast, M., Schaefer, A. W.,Rapoport, I., Kirchhausen, T. and Lemmon, V. (1998). The neural cell adhesion molecule L1 interacts with the AP-2 adaptor and is endocytosed via the clathrin-mediated pathway. J. Neurosci.18,5311-5321.",
      "DOI": "10.1523/JNEUROSCI.18-14-05311.1998"
    },
    {
      "key": "2021042517532726100_REF24",
      "doi-asserted-by": "crossref",
      "unstructured": "Kirchhausen, T. (2002). Clathrin adaptors really adapt. Cell109,413-416.",
      "DOI": "10.1016/S0092-8674(02)00751-1"
    },
    {
      "key": "2021042517532726100_REF25",
      "doi-asserted-by": "crossref",
      "unstructured": "Lamaze, C. and Schmid, S. L. (1995). Recruitment of epidermal growth factor receptors into coated pits requires their activated tyrosine kinase. J. Cell Biol.129, 47-54.",
      "DOI": "10.1083/jcb.129.1.47"
    },
    {
      "key": "2021042517532726100_REF26",
      "doi-asserted-by": "crossref",
      "unstructured": "Lampson, M. A., Schmoranzer, J., Zeigerer, A., Simon, S. M. and McGraw, T. E. (2001). Insulin-regulated release from the endosomal recycling compartment is regulated by budding of specialized vesicles. Mol. Biol. Cell12,3489-3501.",
      "DOI": "10.1091/mbc.12.11.3489"
    },
    {
      "key": "2021042517532726100_REF27",
      "doi-asserted-by": "crossref",
      "unstructured": "McNiven, M. A., Kim, L., Krueger, E. W., Orth, J. D., Cao, H. and Wong, T. W. (2000). Regulated interactions between dynamin and the actin-binding protein cortactin modulate cell shape. J. Cell Biol.151,187-198.",
      "DOI": "10.1083/jcb.151.1.187"
    },
    {
      "key": "2021042517532726100_REF28",
      "doi-asserted-by": "crossref",
      "unstructured": "Merrifield, C. J., Moss, S. E., Ballestrem, C., Imhof, B. A.,Giese, G., Wunderlich, I. and Almers, W. (1999). Endocytic vesicles move at the tips of actin tails in cultured mast cells. Nat. Cell Biol.1,72-74.",
      "DOI": "10.1038/9048"
    },
    {
      "key": "2021042517532726100_REF29",
      "doi-asserted-by": "crossref",
      "unstructured": "Ochoa, G. C., Slepnev, V. I., Neff, L., Ringstad, N., Takei, K.,Daniell, L., Kim, W., Cao, H., McNiven, M., Baron, R. et al.(2000). A functional link between dynamin and the actin cytoskeleton at podosomes. J. Cell Biol.150,377-389.",
      "DOI": "10.1083/jcb.150.2.377"
    },
    {
      "key": "2021042517532726100_REF30",
      "doi-asserted-by": "crossref",
      "unstructured": "Palecek, S. P., Schmidt, C. E., Lauffenburger, D. A. and Horwitz, A. F. (1996). Integrin dynamics on the tail region of migrating fibroblasts. J. Cell Sci.109,941-952.",
      "DOI": "10.1242/jcs.109.5.941"
    },
    {
      "key": "2021042517532726100_REF31",
      "doi-asserted-by": "crossref",
      "unstructured": "Pollard, T. D., Blanchoin, L. and Mullins, R. D.(2000). Molecular mechanisms controlling actin filament dynamics in nonmuscle cells. Annu. Rev. Biophys. Biomol. Struct.29,545-576.",
      "DOI": "10.1146/annurev.biophys.29.1.545"
    },
    {
      "key": "2021042517532726100_REF32",
      "doi-asserted-by": "crossref",
      "unstructured": "Raub, T. J. and Kuentzel, S. L. (1989). Kinetic and morphological evidence for endocytosis of mammalian cell integrin receptors by using an anti-fibronectin receptor beta subunit monoclonal antibody. Exp. Cell Res.184,407-426.",
      "DOI": "10.1016/0014-4827(89)90340-6"
    },
    {
      "key": "2021042517532726100_REF33",
      "doi-asserted-by": "crossref",
      "unstructured": "Sabo, S. L., Ikin, A. F., Buxbaum, J. D. and Greengard, P.(2001). The Alzheimer amyloid precursor protein (APP) and FE65,an APP-binding protein, regulate cell movement. J. Cell Biol.153,1403-1414.",
      "DOI": "10.1083/jcb.153.7.1403"
    },
    {
      "key": "2021042517532726100_REF34",
      "doi-asserted-by": "crossref",
      "unstructured": "Santini, F., Gaidarov, I. and Keen, J. H.(2002). G protein-coupled receptor/arrestin3 modulation of the endocytic machinery. J. Cell Biol.156,665-676.",
      "DOI": "10.1083/jcb.200110132"
    },
    {
      "key": "2021042517532726100_REF35",
      "doi-asserted-by": "crossref",
      "unstructured": "Schmid, S. L. (1997). Clathrin-coated vesicle formation and protein sorting: an integrated process. Annu. Rev. Biochem.66,511-548.",
      "DOI": "10.1146/annurev.biochem.66.1.511"
    },
    {
      "key": "2021042517532726100_REF36",
      "doi-asserted-by": "crossref",
      "unstructured": "Schmid, S. L. and Sorkin, A. D. (2002). Days and knights discussing membrane dynamics in endocytosis: meeting report from the Euresco/EMBL Membrane Dynamics in Endocytosis, 6-11 October in Tomar,Portugal. Traffic3,77-85.",
      "DOI": "10.1046/j.1398-9219.2001.00001.x"
    },
    {
      "key": "2021042517532726100_REF37",
      "doi-asserted-by": "crossref",
      "unstructured": "Schmoranzer, J., Goulian, M., Axelrod, D. and Simon, S. M.(2000). Imaging constitutive exocytosis with total internal reflection fluorescence microscopy. J. Cell Biol.149, 23-32.",
      "DOI": "10.1083/jcb.149.1.23"
    },
    {
      "key": "2021042517532726100_REF38",
      "unstructured": "Sheetz, M. P., Felsenfeld, D., Galbraith, C. G. and Choquet,D. (1999). Cell migration as a five-step cycle. Biochem. Soc. Symp.65,233-243."
    },
    {
      "key": "2021042517532726100_REF39",
      "doi-asserted-by": "crossref",
      "unstructured": "Sieczkarski, S. B. and Whittaker, G. R. (2002). Dissecting virus entry via endocytosis. J. Gen. Virol.83,1535-1545.",
      "DOI": "10.1099/0022-1317-83-7-1535"
    },
    {
      "key": "2021042517532726100_REF40",
      "doi-asserted-by": "crossref",
      "unstructured": "Sun, T. X., van Hoek, A., Huang, Y., Bouley, R., McLaughlin, M. and Brown, D. (2002). Aquaporin-2 localization in clathrin-coated pits: inhibition of endocytosis by dominant-negative dynamin. Am. J. Physiol. Renal Physiol.282,F998-F1011.",
      "DOI": "10.1152/ajprenal.00257.2001"
    },
    {
      "key": "2021042517532726100_REF41",
      "doi-asserted-by": "crossref",
      "unstructured": "Tabb, J. S., Molyneaux, B. J., Cohen, D. L., Kuznetsov, S. A. and Langford, G. M. (1998). Transport of ER vesicles on actin filaments in neurons by myosin V. J. Cell Sci.111,3221-3234.",
      "DOI": "10.1242/jcs.111.21.3221"
    },
    {
      "key": "2021042517532726100_REF42",
      "doi-asserted-by": "crossref",
      "unstructured": "Takei, K. and Haucke, V. (2001). Clathrin-mediated endocytosis: membrane factors pull the trigger. Trends Cell Biol.11,385-391.",
      "DOI": "10.1016/S0962-8924(01)02082-7"
    },
    {
      "key": "2021042517532726100_REF43",
      "doi-asserted-by": "crossref",
      "unstructured": "Takei, K., McPherson, P. S., Schmid, S. L. and de Camilli,P. (1995). Tubular membrane invaginations coated by dynamin rings are induced by GTP-gamma S in nerve terminals. Nature374,186-190.",
      "DOI": "10.1038/374186a0"
    },
    {
      "key": "2021042517532726100_REF44",
      "doi-asserted-by": "crossref",
      "unstructured": "ter Haar, E., Musacchio, A., Harrison, S. C. and Kirchhausen,T. (1998). Atomic structure of clathrin: a beta propeller terminal domain joins an alpha zigzag linker. Cell95,563-573.",
      "DOI": "10.1016/S0092-8674(00)81623-2"
    },
    {
      "key": "2021042517532726100_REF45",
      "doi-asserted-by": "crossref",
      "unstructured": "Tsuboi, T., Terakawa, S., Scalettar, B. A., Fantus, C., Roder,J. and Jeromin, A. (2002). Sweeping model of dynamin activity. Visualization of coupling between exocytosis and endocytosis under an evanescent wave microscope with green fluorescent proteins. J. Biol. Chem.277,15957-15961.",
      "DOI": "10.1074/jbc.C200051200"
    },
    {
      "key": "2021042517532726100_REF46",
      "doi-asserted-by": "crossref",
      "unstructured": "Watanabe, N. and Mitchison, T. J. (2002). Single-molecule speckle analysis of actin filament turnover in lamellipodia. Science295,1083-1086.",
      "DOI": "10.1126/science.1067470"
    }
  ],
  "container-title": "Journal of Cell Science",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "http://journals.biologists.com/jcs/article-pdf/116/5/847/1363358/847.pdf",
      "content-type": "application/pdf",
      "content-version": "vor",
      "intended-application": "syndication"
    },
    {
      "URL": "http://journals.biologists.com/jcs/article-pdf/116/5/847/1363358/847.pdf",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2021,
        4,
        25
      ]
    ],
    "date-time": "2021-04-25T18:10:50Z",
    "timestamp": 1619374250000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://journals.biologists.com/jcs/article/116/5/847/27481/Real-time-analysis-of-clathrin-mediated"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2003,
        3,
        1
      ]
    ]
  },
  "references-count": 46,
  "journal-issue": {
    "issue": "5",
    "published-print": {
      "date-parts": [
        [
          2003,
          3,
          1
        ]
      ]
    }
  },
  "URL": "http://dx.doi.org/10.1242/jcs.00289",
  "relation": {},
  "ISSN": [
    "1477-9137",
    "0021-9533"
  ],
  "subject": [],
  "published": {
    "date-parts": [
      [
        2003,
        3,
        1
      ]
    ]
  }
}