ATP supplementation

Model Organism: Worms Methods & Reagents
1

Hello,

Has any one tried ATP supplementation on worms? Could ATP be taken through feeding?

Thanks.

2

Hi,

ah, at last a biochemistry post…let’s start with your second question first.

Yes you can add ATP as a supplement, but some caveats.

  1. E. coli (and other) bacteria will both release (during log phase) ATP (albeit in nM amounts) and consume it rapidly (during stationary phase, but also if it’s there) . Thus any [ATP] added needs to be assayed to determine just what the concentration is in the worms/medium (before/after).

  2. The peak concentration and concentration profile will vary according to the method of application. Using the ‘mix with live bacteria’ method carries the caveats mentioned above. Dead bacteria would be better, or in liquid culture. I would avoid spotting it onto plates (variable concentration across and through the agar, worms are not always so evenly distributed, say when feeding at the perimetr of the OP50 spread onto your plates.

  3. ATP is stable in buffers (TRIS) and also fairly heat stable (which can be a problem for assays).

Has someone done this with worms? If you haven’t already stumbled across something, then:

Background reading: http://www.wormbook.org/chapters/www_intermetabolism/intermetabolism.html

Of course, the available literature depends on what you want to do? What do you want to do?

Steve

3

Hi Steve,

Thanks for your reply. I’ll probably try adding ATP into agar plate given that ATP is stable in buffer and heat stable. And use killed bacteria to avoid any interference.

I wonder how ATP molecule gets into C. elegans cells? Any transporters responsible for uptaking exogenous ATP? Is it the idea that unless we measure the ATP level inside the worms before/after supplement, we won’t know how much is being taken?

4

Hi,

as far as transport goes, you could start with this review which outlines the possibilities:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2329674/

it’s a much trodden path and there are a number of competing theories.

The point of using the heat-killed bacteria is that they don’t chomp on the ATP or make some themselves. The concentration normally used in the literature is probably way above what you need in the cell so the kinetics will ensure that the concentration the cells of your choice see (within reason) stays constant whilst you are testing the worms.

Do a literature search for the concentrations used generally during incubations (elegans) and also perhaps specifically for the purpose you want to use the ATP for.