title: Characterization of the cytochalasin D-resistant (pinocytic) mechanisms of endocytosis utilized by chlamydiae.
abstract: The cytochalasin D-resistant (pinocytic) portion of the entry of two chlamydia strains (Chlamydia trachomatis L2/434/Bu and Chlamydia psittaci GPIC [guinea pig inclusion conjunctivitis]) was examined. By ultrastructural criteria, few organisms of either strain were observed in association with coated host-cell plasma membrane during entry into McCoy cells; this argues against a coated-pit mechanism of entry. When association with a coated membrane was seen, coat material appeared to pinch off ahead of internalizing chlamydiae. However, entry of both strains was substantially reduced by cytosol acidification, a procedure shown to prevent coated-pit vesiculation (K. Sandvig, S. Olsnes, O. W. Petersen, and B. van Deurs, J. Cell Biol. 105:679-689, 1987). No conclusive evidence of displacement of the fluid-phase marker [3H]sucrose from constitutively forming endocytic vesicles was found. Indeed the entry of strain 434 (but not strain GPIC) was accompanied by the influx of a large volume of fluid, suggesting an inducible mechanism. Additionally, entry of strain 434 (but not strain GPIC) was partially inhibitable by amiloride, yet the drug had no effect on the entry of transferrin, a ligand known to enter solely via coated pits. Our findings endorse the view that chlamydial entry can occur via a pathway involving coated pits. However, the unusual morphology of entry and lack of fluid exclusion are consistent with a process whereby although chlamydiae are not fully enclosed by coat material, their entry is dependent on the vesiculation of coated pits. Furthermore, the data support the proposition that a significant proportion of the entry of strain 434 occurs via an inducible pathway independent of coated-pit uptake.
authors.affiliation: Microbial Molecular Genetics and Cell Biology Group, School of Biological Sciences, University of Birmingham, United Kingdom.
authors.forename: D J
authors.initials: DJ
authors.lastname: Reynolds
authors.forename: J H
authors.initials: JH
authors.lastname: Pearce
mesh_terms: D000643:Ammonium Chloride / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000382:microbiology / Q000502:physiology; D002692:Chlamydia trachomatis / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D002691:Chlamydophila psittaci / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D003034:Coated Pits, Cell-Membrane / Q000382:microbiology / Q000502:physiology; D015638:Cytochalasin D / Q000494:pharmacology*; D003600:Cytosol / Q000187:drug effects; D004352:Drug Resistance, Microbial; D010873:Pinocytosis / Q000187:drug effects / Q000502:physiology*; D014168:Transferrin / Q000378:metabolism
chemical_list: D014168:Transferrin; D000643:Ammonium Chloride; D015638:Cytochalasin D