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Journal of Bacteriology, September 2009, p. 5734-5742, Vol. 191, No. 18
0021-9193/09/$08.00+0     doi:10.1128/JB.00293-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Kinematics of Intracellular Chlamydiae Provide Evidence for Contact-Dependent Development{triangledown} ,{dagger}

David P. Wilson,1 Judith A. Whittum-Hudson,2 Peter Timms,3 and Patrik M. Bavoil4*

National Centre in HIV Epidemiology and Clinical Research, Faculty of Medicine, University of New South Wales, Level 2, 376 Victoria Street, Darlinghurst, Sydney, NSW 2010, Australia,1 Department of Immunology and Microbiology, Wayne State University School of Medicine, 550 East Canfield Avenue, Detroit, Michigan 48201,2 Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove Campus, Blamey Street and Musk Avenue, Brisbane, QLD 4001, Australia,3 Department of Microbial Pathogenesis, University of Maryland Dental School, 650 West Baltimore Street, Baltimore, Maryland 212014

Received 4 March 2009/ Accepted 5 June 2009

A crucial process of chlamydial development involves differentiation of the replicative reticulate body (RB) into the infectious elementary body (EB). We present experimental evidence to provide support for a contact-dependent hypothesis for explaining the trigger involved in differentiation. We recorded live-imaging of Chlamydia trachomatis-infected McCoy cells at key times during development and tracked the temporospatial trajectories of individual chlamydial particles. We found that movement of the particles is related to development. Early to mid-developmental stages involved slight wobbling of RBs. The average speed of particles increased sharply at 24 h postinfection (after the estimated onset of RB to EB differentiation). We also investigated a penicillin-supplemented culture containing EBs, RBs, and aberrantly enlarged, stressed chlamydiae. Near-immobile enlarged particles are consistent with their continued tethering to the chlamydial inclusion membrane (CIM). We found a significantly negative, nonlinear association between speed and size/type of particles, providing further support for the hypothesis that particles become untethered near the onset of RB to EB differentiation. This study establishes the relationship between the motion properties of the chlamydiae and developmental stages, whereby wobbling RBs gradually lose contact with the CIM, and RB detachment from the CIM is coincidental with the onset of late differentiation.

* Corresponding author. Mailing address: Department of Microbial Pathogenesis, University of Maryland Dental School, 650 W. Baltimore St., Baltimore, MD 21201. Phone: (410) 706-6789. Fax: (410) 706-0865. E-mail: pbavoil{at}umaryland.edu

{triangledown} Published ahead of print on 19 June 2009.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, September 2009, p. 5734-5742, Vol. 191, No. 18
0021-9193/09/$08.00+0     doi:10.1128/JB.00293-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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