Three-Dimensional Observations of an Aperiodic Oscillatory Gliding Behavior in <named-content content-type="genus-species">Myxococcus xanthus</named-content> Using Confocal Interference Reflection Microscopy

ABSTRACT The deltaproteobacterium Myxococcus xanthus is a model for bacterial motility and has provided unprecedented insights into bacterial swarming behaviors. Fluorescence microscopy techniques have been invaluable in defining the mechanisms that are involved in gliding motility, but these have a...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Liam M. Rooney, Lisa S. Kölln, Ross Scrimgeour, William B. Amos, Paul A. Hoskisson, Gail McConnell
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	3D imaging bacterial motility gliding motility label free live-cell imaging optical microscopy Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/f23be4bc99694325be4cf1ca5b8e6bd2
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:f23be4bc99694325be4cf1ca5b8e6bd2
record_format	dspace
spelling	oai:doaj.org-article:f23be4bc99694325be4cf1ca5b8e6bd22021-11-15T15:27:52ZThree-Dimensional Observations of an Aperiodic Oscillatory Gliding Behavior in <named-content content-type="genus-species">Myxococcus xanthus</named-content> Using Confocal Interference Reflection Microscopy10.1128/mSphere.00846-192379-5042https://doaj.org/article/f23be4bc99694325be4cf1ca5b8e6bd22020-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00846-19https://doaj.org/toc/2379-5042ABSTRACT The deltaproteobacterium Myxococcus xanthus is a model for bacterial motility and has provided unprecedented insights into bacterial swarming behaviors. Fluorescence microscopy techniques have been invaluable in defining the mechanisms that are involved in gliding motility, but these have almost entirely been limited to two-dimensional (2D) studies, and there is currently no understanding of gliding motility in a three-dimensional (3D) context. We present here the first use of confocal interference reflection microscopy (IRM) to study gliding bacteria, revealing aperiodic oscillatory behavior with changes in the position of the basal membrane relative to the substrate on the order of 90 nm in vitro. First, we use a model planoconvex lens specimen to show how topological information can be obtained from the wavelength-dependent interference pattern in IRM. We then use IRM to observe gliding M. xanthus bacteria and show that cells undergo previously unobserved changes in their adhesion profile as they glide. We compare the wild type with mutants that have reduced motility, which also exhibit the same changes in the adhesion profile during gliding. We find that the general gliding behavior is independent of the proton motive force-generating complex AglRQS and suggest that the novel behavior that we present here may be a result of recoil and force transmission along the length of the cell body following firing of the type IV pili. IMPORTANCE 3D imaging of live bacteria with optical microscopy techniques is a challenge due to the small size of bacterial cells, meaning that previous studies have been limited to observing motility behavior in 2D. We introduce the application of confocal multiwavelength interference reflection microscopy to bacteria, which enables visualization of 3D motility behaviors in a single 2D image. Using the model organism Myxococcus xanthus, we identified novel motility behaviors that are not explained by current motility models, where gliding bacteria exhibit aperiodic changes in their adhesion to an underlying solid surface. We concluded that the 3D behavior was not linked to canonical motility mechanisms and that IRM could be applied to study a range of microbiological specimens with minimal adaptation to a commercial microscope.Liam M. RooneyLisa S. KöllnRoss ScrimgeourWilliam B. AmosPaul A. HoskissonGail McConnellAmerican Society for Microbiologyarticle3D imagingbacterial motilitygliding motilitylabel freelive-cell imagingoptical microscopyMicrobiologyQR1-502ENmSphere, Vol 5, Iss 1 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	3D imaging bacterial motility gliding motility label free live-cell imaging optical microscopy Microbiology QR1-502
spellingShingle	3D imaging bacterial motility gliding motility label free live-cell imaging optical microscopy Microbiology QR1-502 Liam M. Rooney Lisa S. Kölln Ross Scrimgeour William B. Amos Paul A. Hoskisson Gail McConnell Three-Dimensional Observations of an Aperiodic Oscillatory Gliding Behavior in <named-content content-type="genus-species">Myxococcus xanthus</named-content> Using Confocal Interference Reflection Microscopy
description	ABSTRACT The deltaproteobacterium Myxococcus xanthus is a model for bacterial motility and has provided unprecedented insights into bacterial swarming behaviors. Fluorescence microscopy techniques have been invaluable in defining the mechanisms that are involved in gliding motility, but these have almost entirely been limited to two-dimensional (2D) studies, and there is currently no understanding of gliding motility in a three-dimensional (3D) context. We present here the first use of confocal interference reflection microscopy (IRM) to study gliding bacteria, revealing aperiodic oscillatory behavior with changes in the position of the basal membrane relative to the substrate on the order of 90 nm in vitro. First, we use a model planoconvex lens specimen to show how topological information can be obtained from the wavelength-dependent interference pattern in IRM. We then use IRM to observe gliding M. xanthus bacteria and show that cells undergo previously unobserved changes in their adhesion profile as they glide. We compare the wild type with mutants that have reduced motility, which also exhibit the same changes in the adhesion profile during gliding. We find that the general gliding behavior is independent of the proton motive force-generating complex AglRQS and suggest that the novel behavior that we present here may be a result of recoil and force transmission along the length of the cell body following firing of the type IV pili. IMPORTANCE 3D imaging of live bacteria with optical microscopy techniques is a challenge due to the small size of bacterial cells, meaning that previous studies have been limited to observing motility behavior in 2D. We introduce the application of confocal multiwavelength interference reflection microscopy to bacteria, which enables visualization of 3D motility behaviors in a single 2D image. Using the model organism Myxococcus xanthus, we identified novel motility behaviors that are not explained by current motility models, where gliding bacteria exhibit aperiodic changes in their adhesion to an underlying solid surface. We concluded that the 3D behavior was not linked to canonical motility mechanisms and that IRM could be applied to study a range of microbiological specimens with minimal adaptation to a commercial microscope.
format	article
author	Liam M. Rooney Lisa S. Kölln Ross Scrimgeour William B. Amos Paul A. Hoskisson Gail McConnell
author_facet	Liam M. Rooney Lisa S. Kölln Ross Scrimgeour William B. Amos Paul A. Hoskisson Gail McConnell
author_sort	Liam M. Rooney
title	Three-Dimensional Observations of an Aperiodic Oscillatory Gliding Behavior in <named-content content-type="genus-species">Myxococcus xanthus</named-content> Using Confocal Interference Reflection Microscopy
title_short	Three-Dimensional Observations of an Aperiodic Oscillatory Gliding Behavior in <named-content content-type="genus-species">Myxococcus xanthus</named-content> Using Confocal Interference Reflection Microscopy
title_full	Three-Dimensional Observations of an Aperiodic Oscillatory Gliding Behavior in <named-content content-type="genus-species">Myxococcus xanthus</named-content> Using Confocal Interference Reflection Microscopy
title_fullStr	Three-Dimensional Observations of an Aperiodic Oscillatory Gliding Behavior in <named-content content-type="genus-species">Myxococcus xanthus</named-content> Using Confocal Interference Reflection Microscopy
title_full_unstemmed	Three-Dimensional Observations of an Aperiodic Oscillatory Gliding Behavior in <named-content content-type="genus-species">Myxococcus xanthus</named-content> Using Confocal Interference Reflection Microscopy
title_sort	three-dimensional observations of an aperiodic oscillatory gliding behavior in <named-content content-type="genus-species">myxococcus xanthus</named-content> using confocal interference reflection microscopy
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/f23be4bc99694325be4cf1ca5b8e6bd2
work_keys_str_mv	AT liammrooney threedimensionalobservationsofanaperiodicoscillatoryglidingbehaviorinnamedcontentcontenttypegenusspeciesmyxococcusxanthusnamedcontentusingconfocalinterferencereflectionmicroscopy AT lisaskolln threedimensionalobservationsofanaperiodicoscillatoryglidingbehaviorinnamedcontentcontenttypegenusspeciesmyxococcusxanthusnamedcontentusingconfocalinterferencereflectionmicroscopy AT rossscrimgeour threedimensionalobservationsofanaperiodicoscillatoryglidingbehaviorinnamedcontentcontenttypegenusspeciesmyxococcusxanthusnamedcontentusingconfocalinterferencereflectionmicroscopy AT williambamos threedimensionalobservationsofanaperiodicoscillatoryglidingbehaviorinnamedcontentcontenttypegenusspeciesmyxococcusxanthusnamedcontentusingconfocalinterferencereflectionmicroscopy AT paulahoskisson threedimensionalobservationsofanaperiodicoscillatoryglidingbehaviorinnamedcontentcontenttypegenusspeciesmyxococcusxanthusnamedcontentusingconfocalinterferencereflectionmicroscopy AT gailmcconnell threedimensionalobservationsofanaperiodicoscillatoryglidingbehaviorinnamedcontentcontenttypegenusspeciesmyxococcusxanthusnamedcontentusingconfocalinterferencereflectionmicroscopy
_version_	1718427984936828928

Three-Dimensional Observations of an Aperiodic Oscillatory Gliding Behavior in <named-content content-type="genus-species">Myxococcus xanthus</named-content> Using Confocal Interference Reflection Microscopy

Ejemplares similares