We want to culture hippocampal slices and we want to selectively deplete astrocytes and oligodendrocytes without affecting the neurons. We know how to deplete microglial cells and it works more or less effectively. Is there any compound (like clodronate) to deplete these cells pharmacological? We found some papers describing methods but I don't think that they are applicable in our experiments. Thanks for your answers =)!
The attached Paper"Brain Slices as Models for Neurodegenerative Disease and Screening Platforms to Identify Novel Therapeutics"
please check out.
Best wishes
Hi Natascha,
Have a look at the attached paper 'Glutamate released spontaneously from astrocytes sets the threshold for synaptic plasticity' by Bonansco et al.
The authors use fluorocitrate, a reversible metabolic gliotoxin (aconitase inhibitor). Seems to work pretty well :)
Good luck,
Marcin
Thank you very much for your quick responses! Ali, thank you for the paper recommendation, I will have a look at this :).
I already found a paper where they injected D,L-fluorocitic acid into living animals, but this is too complicated for our set up. But I will have a look at the paper you recommended, Marcin =)!
Thank you Alexander, too, I will have a look at this toxin :).
I use fluoroacetate or fluorocitrate. These gliotoxins inhibit the kreb cyle of astrocytes by entering the cell using an acetate transporter, which is mainly in astrocytes. Fonnum et al., 1997 explained that oligodendrocytes are also affected by these gliotoxins.
In addition to fluorocitrate you can try another gliotoxin: amino adipic acid. It damages astrocytes, and possiblly also oligodendrocytes. See: Aminoadipic acid toxic effects on retinal glial cells. Ishikawa Y, Mine S. Jpn J Ophthalmol. 1983;27(1):107-18
Since fluorocitrate is often described as reversible, has anyone observed the washout of the toxin in slices? If so, what is the time course?
Hello again, just a brief update. We now considered to use toxin-coupled antibodies to deplete oligodendrocytes from our slices and we will also try to do this for strocytes. I let you know when it works ;). Thanks for all the answers so far =)
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