Nanotubes

Nanotubes

Just as people, bacteria need to communicate with others. For this goal there are many paths, and one of the most recent discoveries are Nanotubes that bridge neighboring cells, providing a network for exchange of cellular molecules within, and between species. Ben-Yehuda et al discovered these nanotubes in 2011. They show an extraordinary form of communication between Bacillus subtilis. Our team was astonished because of the implications. In the article they described that GFP and calcein, two molecules which cannot leave the cytoplasm, can be transferred to neighboring cells in B. subtilis. This means bacterium can share cytoplasm. The complex network of cells sharing cytoplasm that can be created was our main motivation to create Bacillus booleanus.

Little is still known about nanotubes, and that's what makes them a very interesting subject of study. We are not the first iGEM team interested in working with them. In 2011 the Paris_Bettencourt team worked with nanotubes, their goal being to characterize them

To accomplish our goal of communicate our logic gates, we contacted Ben Yehuda group, and Paris Bettencourt iGEM 2011 team, to obtain protocols and work experience.

We worked to recreate the formation of nanotubes, and we did it!

PY79 cells were grown to midexponential phase, plated on LB agar, incubated for 6 hr at 37°C, and visualized by SEM Rej34 circles: (A) Bacillus subtilis cells(x 7,500). The red circles indicate intercellular nanotubes connecting neighboring cells. The scale bar represents 1 micrometer.

We prepare our cells with the SEM Analysis Protocol, We fixed our cells in the right moment (6 hrs of growth at 37°C), that is when they are in the exponential growth phase, and then we watch them in the SEM (Scanning Electron Microscope) and we get different and amazing pictures.

We got similar results to the article, tube length ranged from 0.25 micrometers to 0.9 micrometers, whereas width ranged approximately from 50 to 80 nm.

PY79 cells were grown to midexponential phase, plated on LB agar, incubated for 6 hr at 37°C, and visualized by SEM Rej35_valores:(A)Measurment of the length and width of the nanotubes connecting the (PY79) cells grown on solid LB medium. The scale bar represent 1 micrometer

We also got an amazing picture about the nanotubes in which we can see, that there are many nanotubes among the cells, and that they communicate between different nanotubes to the same cell, observing this amount of nanotubes, we could think that the probability of transfer will raise.

PY79 cells were grown to midexponential phase, plated on LB agar, incubated for 6 hr at 37°C, and visualized by SEM Rej48: (A) Field of cells demonstrating the occurrence of a network of intercellular nanotubes (X 10,000). The scale bar represents 1 micrometer

Slider captions
PY79 cells were grown to midexponential phase, plated on LB agar, incubated for 6 hr at 37°C, and visualized by SEM Rej43, 44, 46, 47.

Please see our wetlab notebook in the clicking the following image:

Bacillus Notebook