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Identification of vacuoles containing extraintestinal differentiated forms of Legionella pneumophila in colonized Caenorhabditis elegans soil nematodes.

Hellinga JR, Garduño RA, Kormish JD, Tanner JR, Khan D, Buchko K, Jimenez C, Pinette MM, Brassinga AK - Microbiologyopen (2015)

Bottom Line: A previous study employing the metazoan Caenorhabditis elegans, a member of the Rhabditidae family of free-living soil nematodes, demonstrated that the intestinal lumen can be colonized with L. pneumophila.While both replicative forms and differentiated forms were observed in C. elegans, these morphologically distinct forms were initially observed to be restricted to the intestinal lumen.Furthermore, establishment of LCVs in the gonadal tissue was Dot/Icm dependent and required the presence of the endocytic factor RME-1 to gain access to maturing oocytes.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Faculty of Science, University of Manitoba, Winnipeg, Manitoba, Canada, R3T 2N2.

No MeSH data available.


Related in: MedlinePlus

Intestinal and extra-intestinal locations of GFP-tagged Legionella pneumophila in Caenorhabditis elegans. Microscopic images of glo-3(kx94) nematodes fed with Lp02 PmagA::gfp strain for (A) 5 days and (B) 6 days. Note the fluorescent rod-shaped bacteria (white arrow) in the intestine in (A) and forming a highly fluorescent bacterial mass in the intestine in (B). Note also the fluorescent (white arrows) and nonfluorescent (white arrowheads) coccoid-shaped bacteria in vacuoles (i.e., Legionella-containing vacuoles) in (B) of which only one is visible due to differing focal planes. Panels (i–iii) represent DIC, green and merge channels. Scale bar is 5 μm. Still images in (B) taken from Video S5.
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fig03: Intestinal and extra-intestinal locations of GFP-tagged Legionella pneumophila in Caenorhabditis elegans. Microscopic images of glo-3(kx94) nematodes fed with Lp02 PmagA::gfp strain for (A) 5 days and (B) 6 days. Note the fluorescent rod-shaped bacteria (white arrow) in the intestine in (A) and forming a highly fluorescent bacterial mass in the intestine in (B). Note also the fluorescent (white arrows) and nonfluorescent (white arrowheads) coccoid-shaped bacteria in vacuoles (i.e., Legionella-containing vacuoles) in (B) of which only one is visible due to differing focal planes. Panels (i–iii) represent DIC, green and merge channels. Scale bar is 5 μm. Still images in (B) taken from Video S5.

Mentions: Bacterial colonization of the intestinal tract of glo-3(kx94) nematodes with Lp02 PmagA::gfpmut3 proceeded as observed with Lp02 in this study but as expected with a fluorescence intensity lower than previously observed with the plasmid-borne fluorescent protein expression of colonizing Lp02 in the Brassinga et al. (2010) study (Fig.3A). Vacuoles containing motile coccoid-shaped bacterial forms found in the pseudocoelomic cavity (data not shown) and gonadal tissue (Fig.3B, Video S5) were also fluorescent in the green channel confirming the identity of these bacterial forms as Lp02. It should be noted that only some, but not all of the motile coccoid-shaped bacterial forms, were fluorescent suggesting that the cyst forms became metabolically dormant and no longer expressed GFP protein upon completion of cyst biogenesis as detailed in Hiltz et al. (2004) (Fig.3B, Video S5). The vacuoles containing the motile coccoid-shaped bacterial forms, henceforth referred to as LCVs, are distinct in size, motility, and structure when compared to sperm contained within the spermatheca section of the C. elegans reproductive tract (Video S6).


Identification of vacuoles containing extraintestinal differentiated forms of Legionella pneumophila in colonized Caenorhabditis elegans soil nematodes.

Hellinga JR, Garduño RA, Kormish JD, Tanner JR, Khan D, Buchko K, Jimenez C, Pinette MM, Brassinga AK - Microbiologyopen (2015)

Intestinal and extra-intestinal locations of GFP-tagged Legionella pneumophila in Caenorhabditis elegans. Microscopic images of glo-3(kx94) nematodes fed with Lp02 PmagA::gfp strain for (A) 5 days and (B) 6 days. Note the fluorescent rod-shaped bacteria (white arrow) in the intestine in (A) and forming a highly fluorescent bacterial mass in the intestine in (B). Note also the fluorescent (white arrows) and nonfluorescent (white arrowheads) coccoid-shaped bacteria in vacuoles (i.e., Legionella-containing vacuoles) in (B) of which only one is visible due to differing focal planes. Panels (i–iii) represent DIC, green and merge channels. Scale bar is 5 μm. Still images in (B) taken from Video S5.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4554460&req=5

fig03: Intestinal and extra-intestinal locations of GFP-tagged Legionella pneumophila in Caenorhabditis elegans. Microscopic images of glo-3(kx94) nematodes fed with Lp02 PmagA::gfp strain for (A) 5 days and (B) 6 days. Note the fluorescent rod-shaped bacteria (white arrow) in the intestine in (A) and forming a highly fluorescent bacterial mass in the intestine in (B). Note also the fluorescent (white arrows) and nonfluorescent (white arrowheads) coccoid-shaped bacteria in vacuoles (i.e., Legionella-containing vacuoles) in (B) of which only one is visible due to differing focal planes. Panels (i–iii) represent DIC, green and merge channels. Scale bar is 5 μm. Still images in (B) taken from Video S5.
Mentions: Bacterial colonization of the intestinal tract of glo-3(kx94) nematodes with Lp02 PmagA::gfpmut3 proceeded as observed with Lp02 in this study but as expected with a fluorescence intensity lower than previously observed with the plasmid-borne fluorescent protein expression of colonizing Lp02 in the Brassinga et al. (2010) study (Fig.3A). Vacuoles containing motile coccoid-shaped bacterial forms found in the pseudocoelomic cavity (data not shown) and gonadal tissue (Fig.3B, Video S5) were also fluorescent in the green channel confirming the identity of these bacterial forms as Lp02. It should be noted that only some, but not all of the motile coccoid-shaped bacterial forms, were fluorescent suggesting that the cyst forms became metabolically dormant and no longer expressed GFP protein upon completion of cyst biogenesis as detailed in Hiltz et al. (2004) (Fig.3B, Video S5). The vacuoles containing the motile coccoid-shaped bacterial forms, henceforth referred to as LCVs, are distinct in size, motility, and structure when compared to sperm contained within the spermatheca section of the C. elegans reproductive tract (Video S6).

Bottom Line: A previous study employing the metazoan Caenorhabditis elegans, a member of the Rhabditidae family of free-living soil nematodes, demonstrated that the intestinal lumen can be colonized with L. pneumophila.While both replicative forms and differentiated forms were observed in C. elegans, these morphologically distinct forms were initially observed to be restricted to the intestinal lumen.Furthermore, establishment of LCVs in the gonadal tissue was Dot/Icm dependent and required the presence of the endocytic factor RME-1 to gain access to maturing oocytes.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Faculty of Science, University of Manitoba, Winnipeg, Manitoba, Canada, R3T 2N2.

No MeSH data available.


Related in: MedlinePlus