We aim to prepare and stimulate acute brain Slices from different fish species. Does anyone know if this has been done before?
Very interesting Ida! The procedures of slice preparation should not grossly differ from those used for cutting mammalian brains. I have some experience with recordings from hemisected spinal cord of amphibians and reptiles with sharp electrodes. Please check old papers (1960-1980), I know there are a few but detailed studies with the use of sharp electrodes. Having said that, defining brain parts and compartments in fish homologous to those of mammalian could be really challenging. You will be limited to work with the brain stem, cerebellum and mid-brain structures, which are anatomically messy and functionally poorly differentiated. Have fun and good luck! SV
Thank you very much for your answer, Saak! Luckily, the brain stem is probably our area of interesst since the parasite we are investigating infests this area. I thought perhaps we would have to use a vibratome or something to Slice the brain, but I will check out papers where they have used electrodes.
Note that electrodes have been used for recording electrical activity from neurons not for slice preparation. I think you better stick with vibratome for cutting slices...
Depending on the species you will need to adjust the pH and osmolality. In general there is a increase in pH with decreasing temperature.12 degree C pH 7.9-8 and at 26 degree C 7.75. In addition, if you use fresh water species the osmolality is usually 280-300 mOsm while salt water species have around 320 mOsm. During slicing you should avoid Ca2+, and have reduced Na+ levels. You can also try NMDG based solution during recovery similar to the one described at http://www.brainslicemethods.com. If possible, you can also try whole brain preparation. We use whole brain when performing patch-clamp recordings from neurones located in the telencephalon. See Nancy Wayne et al. http://www.jove.com/video/50312/recording-electrical-activity-from-identified-neurons-intact-brain.
Best of luck:)
Dear Ida,
In addition to wonderful comments above, if you decide to go ahead with vibratom slices, have a look at my methods paper below for detailed description of how to cut brain slices in mice. You may need the modify the extracellular solution and some of the procedures as suggested by Kjetil, but the general principles should be pretty much the same.
Best wishes, Refik
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