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Time-lapse microscopy of surface translocation by P. gingivalis strain 381 and the Δ ppad mutant. (a) Schematic illustration representing sequential stages of surface translocation by reference strain 381 as recorded for more than 160 h of incubation using the chamber slide system. (b) Frames 1a to e show the chronology of surface translocation by reference strain 381, including the hydrating stage and the formation of moving pseudofilaments, outward spreading of the colony, and subdiffusive cell-driven motility by individual cells that progresses to surface translocation and the formation of distal <t>biofilms</t> over time. Frames 2a to f represent the recordings of disordered stages of surface translocation by strain 381Δ ppad . The absence of ppad resulted in the formation of a limited hydration zone but very long pseudofilaments whose aggregation resulted in the observation of scattered biofilms and the extension of the time required to see outward spreading of the cells and subdiffusive cell-driven motility. Scale bars: 10 μm.
Baphk, supplied by Northeast Laboratory Services, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Time-lapse microscopy of surface translocation by P. gingivalis strain 381 and the Δ ppad mutant. (a) Schematic illustration representing sequential stages of surface translocation by reference strain 381 as recorded for more than 160 h of incubation using the chamber slide system. (b) Frames 1a to e show the chronology of surface translocation by reference strain 381, including the hydrating stage and the formation of moving pseudofilaments, outward spreading of the colony, and subdiffusive cell-driven motility by individual cells that progresses to surface translocation and the formation of distal biofilms over time. Frames 2a to f represent the recordings of disordered stages of surface translocation by strain 381Δ ppad . The absence of ppad resulted in the formation of a limited hydration zone but very long pseudofilaments whose aggregation resulted in the observation of scattered biofilms and the extension of the time required to see outward spreading of the cells and subdiffusive cell-driven motility. Scale bars: 10 μm.

Journal: Journal of Bacteriology

Article Title: PPAD Activity Promotes Outer Membrane Vesicle Biogenesis and Surface Translocation by Porphyromonas gingivalis

doi: 10.1128/JB.00343-20

Figure Lengend Snippet: Time-lapse microscopy of surface translocation by P. gingivalis strain 381 and the Δ ppad mutant. (a) Schematic illustration representing sequential stages of surface translocation by reference strain 381 as recorded for more than 160 h of incubation using the chamber slide system. (b) Frames 1a to e show the chronology of surface translocation by reference strain 381, including the hydrating stage and the formation of moving pseudofilaments, outward spreading of the colony, and subdiffusive cell-driven motility by individual cells that progresses to surface translocation and the formation of distal biofilms over time. Frames 2a to f represent the recordings of disordered stages of surface translocation by strain 381Δ ppad . The absence of ppad resulted in the formation of a limited hydration zone but very long pseudofilaments whose aggregation resulted in the observation of scattered biofilms and the extension of the time required to see outward spreading of the cells and subdiffusive cell-driven motility. Scale bars: 10 μm.

Article Snippet: To determine if PPAD activity alters gene expression, we performed transcriptomic analysis on RNA extracted from cells of strains 381 and 381Δ ppad grown as colony biofilms on BAPHK (Northeast Laboratory Services) for 24 h (the same growth conditions as used for cryo-SEM imaging).

Techniques: Time-lapse Microscopy, Translocation Assay, Mutagenesis, Incubation

Deletion of ppad inhibits OMV biogenesis and release. (a) NanoSight analysis of crude OMV preparations from 381 and 381Δ ppad planktonic cultures. The 381Δ ppad mutant produced OMVs that were fewer and smaller in size than those produced by strain 381. (b) Cryo-SEM imaging of colony biofilms showed that the 381 ppad mutant produced few to no OMVs within the biofilm matrix compared to the parent strain. (Inlay) Magnified section of micrograph. Scale bar: 2 μm. (c) Formation of biopearling assemblies by strain 381. SEM imaging of fixed content of the hydration zone showed that strain 381 formed biopearling assemblies (yellow arrows) via connecting membrane protrusions and large-size OMVs (red arrows) under surface translocation conditions. Deletion of ppad resulted in production of few small-sized OMVs (pink arrows) and in inhibition of biogenesis of large-size OMVs and biopearling assemblies. (Inlay) Magnified section of micrograph. Scale bar: 3 μm.

Journal: Journal of Bacteriology

Article Title: PPAD Activity Promotes Outer Membrane Vesicle Biogenesis and Surface Translocation by Porphyromonas gingivalis

doi: 10.1128/JB.00343-20

Figure Lengend Snippet: Deletion of ppad inhibits OMV biogenesis and release. (a) NanoSight analysis of crude OMV preparations from 381 and 381Δ ppad planktonic cultures. The 381Δ ppad mutant produced OMVs that were fewer and smaller in size than those produced by strain 381. (b) Cryo-SEM imaging of colony biofilms showed that the 381 ppad mutant produced few to no OMVs within the biofilm matrix compared to the parent strain. (Inlay) Magnified section of micrograph. Scale bar: 2 μm. (c) Formation of biopearling assemblies by strain 381. SEM imaging of fixed content of the hydration zone showed that strain 381 formed biopearling assemblies (yellow arrows) via connecting membrane protrusions and large-size OMVs (red arrows) under surface translocation conditions. Deletion of ppad resulted in production of few small-sized OMVs (pink arrows) and in inhibition of biogenesis of large-size OMVs and biopearling assemblies. (Inlay) Magnified section of micrograph. Scale bar: 3 μm.

Article Snippet: To determine if PPAD activity alters gene expression, we performed transcriptomic analysis on RNA extracted from cells of strains 381 and 381Δ ppad grown as colony biofilms on BAPHK (Northeast Laboratory Services) for 24 h (the same growth conditions as used for cryo-SEM imaging).

Techniques: Mutagenesis, Produced, Imaging, Membrane, Translocation Assay, Inhibition