lieratura_22.11.11.pptVIP

Intercellular Nanotubes Mediate Bacterial Communication Gyanendra P. Dubey and Sigal Ben-Yehuda Department of Microbiology and Molecular Genetics, Institute for Medical Research Israel-Canada (IMRIC), The Hebrew University-Hadassah Medical School, POB 12272, The Hebrew University of Jerusalem, 91120 Jerusalem, Israel 144, 590-600, February 18, 2011 Nanotubular channels formed among bacterial cells grown on solid surface, specifically induced under this growth condition connect bacteria of the same and different species effective conduit for exchange of intracellular content transfer of cytoplasmic fluorescent molecules between adjacent cells molecular exchange enables cells to transiently aquire nonhereditary antibiotic resistance nonconjugative plasmids could be transferred thereby conferring hereditary features to recipient cells Neighboring B. subtilis cells exchange cytoplasmic molecules a molecular gradient between neighboring B. subtilis cells: - SB444 (gfp+) spotted on solid medium alongside PY79 (gfp-) - grow for 15 hr - visualized by fluorescence microscopy a green fluorescence gradient was observed to emanate from the gfp+ cells toward the gfp- cells fluorescence gradient was associated exclusively with the presence of cells Average fluorescence intensity of the gfp- population as a function of the distance from gfp+ population. GFP gradient covered a distance of approximately 40 μm Time-lapse fluorescence microscopy t0min t30min t60min exponentially growing gfp- and gfp+ cells were mixed, plated on an LB plates and incubated at 37?C phase contrast and fluorescence images collected at 10 min intervals Transfer of fluorescent molecules between neighboring cells is increased over time. Transfer of calcein between neighboring B. subtilis cells calcein is nongenetically encoded cytoplasmic fluorophore – nonfluorescent acetoxymethylester (AM),that can traverse cell membrane. After passage into the cytoplasm is hydrolysed by endogen