title: Tracking Differential Endocytosis and Trafficking of Semaphorin Receptor Complexes in Responding Nerve Growth Cones.
abstract: The study of receptor endocytosis is important to our understanding of the signal transduction events initiated by axon guidance cues in growth cones. Fab fragments of antibodies to guidance receptors and endocytic cargoes like transferrin and cholera toxin-B are the tools of choice for studying the dynamics of receptor internalization and intracellular trafficking by different pathways. We describe a method where in vitro cultures of growth cones are incubated with these ligands in the presence or absence of Sema3A, followed by stripping of remaining ligand on cell-surface and analysis by immunofluorescence techniques. These techniques can be employed for studying the endocytosis of any axon guidance receptor in response to attractive or repulsive guidance cues and, in particular, to allow the differential trafficking of specific receptor components to be revealed.
authors: Department of Biomedical Science, University of Sheffield, Sheffield, S10 2TN, UK. puneetd@mail.med.upenn.edu.|Department of Neuroscience, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA. puneetd@mail.med.upenn.edu.
authors: Puneet
authors: P
authors: Dang
authors: Department of Biomedical Science, University of Sheffield, Sheffield, S10 2TN, UK.
authors: Andrew J W
authors: AJ
authors: Furley
mesh_terms: D001692:Biological Transport; D004705:Endocytosis; D005455:Fluorescent Antibody Technique; D005727:Ganglia, Spinal / Q000166:cytology; D020439:Growth Cones / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D066298:In Vitro Techniques; D008024:Ligands; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D040121:Semaphorin-3A / Q000378:metabolism*
chemical_list: D008024:Ligands; D009419:Nerve Tissue Proteins; C492119:PLXNB1 protein, human; D011956:Receptors, Cell Surface; D040121:Semaphorin-3A
keywords: Clathrin-independent endocytosis; Colocalization; Endosome; Intracellular trafficking; Semaphorin