I agree with Dr Kohling . However, I would also further emphasize that the thickness of the slice, the type of recording chamber, whether you are using a submerged slice or a slice that is at the interface are all critical variables that will affect pO2 tension. Also how do you monitor the health of the slice before OGD? Do all the EPSPs have max amplitudes or is there a lot of variation. These variables all make a difference.

Yeah. Both answers will improve your research.

Dear Mr. Wang,

on the recovery:

Additional to anoxic depolarization, this paper ( http://www.ncbi.nlm.nih.gov/pubmed/22864185 ) shows that a build-up of Adenosine in the extracellular space (due to breakdown of ATP) halts synaptic functioning. This effect remains for a few minutes after the repolarization that follows reoxygenation.

on failure of recovery:

This is a (clinical) review that discusses selective synaptic damage: http://www.ncbi.nlm.nih.gov/pubmed/22207505

I hope this is useful for your research.

I'm really appreciative of the great advices from all of you!!! For OGD, ACSF with sucrose instead of glucose was bubbled with 95% N2+5% CO2 for more than 25 minutes before perfusion (and keep bubbling all the time; flow rate is ~2ml/min). I used two kinds of chambers, one is RC-26G from Warner Instruments, and another one is a Tissue recording chamber(2 channel) from Automate Scientific Inc. At the begining, I noticed the different between chambers, so I adjust the ACSF solution surface level to ensure that slices were completly immersed into ACSF in both chambers. I also get the prety same slope for fEPSPs supression (from 100% to~ 0%) from both chambers, and normally it took ~15 minutes. I choose the slices (at 400um) that has good fEPSPs responds for OGD experiments, usually, the quality of my slices are great.

with this experiment setup, would it be acceptable to state that my slices were really insulted by OGD? Should I measure the pO2 tension in my codition, and what equipment I need to have it tested?

Dear Wang,

thanks for your contact.

It is not my research focus. If you can, click in link bellow to you know our projects. Maybe we can do something together.

http://www.nucleofutebol.ufv.br/nucleofutebol_ingles/index.php

Best Regards.

Oops~ Thanks so much for your message, Israel! Probably you want to see how it will workfor spoters if you force mitos work hard to produce more energy

So far nobody mentioned that also a variation in temperature could explain differences in recovery ...

Thanks, Reymann. I used the temperature contoller to hold the ACSF at 28C.

We used 400 µm slices and 34-35C (!):

60 min after 5 min hypoxia we observe a 50% recovery of PS, Best, KR

Thanks, Klaus. I tried more slices, and 40 min after 30 min OGD I still observe a 100% recovery. I need figure out what is the matter here in my experiments

Well the experts must understand, but to me OGD is just an unexplained TLA...

Oxygen and glucose deprivation. (You made me smile today after I googled TLA.)

Thanks! There is a story about an expert in computer science (but it could apply to any branch of science and technology) being asked "What do you think is the biggest barrier to progress in your field". His answer: "There are only 17000 possible TLAs..."

Sorry - very off-topic...

Thanks for your interests, Gavin. I also googled TLA and looks like it stands for many things. But the story you tell is very typical