I want to test the effect of dopamine in fish neuron by electrophysiology. Yet, dopamine is oxidised quickly, making it impossible to use it during stimulation and recording.
In molecular biology, people often add DTT in their solutions to avoid oxidation of the reagents. Is there anything I could add to my dopamine solution to avoid its oxidation?
I believe many people use ascorbic acid to prevent oxidation of dopamine in these type of experiments.
Ascorbate is OK but sodium metabisulfate is a better antioxidant for DA
Note that ascorbate is a subunit-selective (Cav3.2) antagonist for rodent and human T-type calcium channels -- IC50 in the 10microM range. Not sure if fish express analogous isoforms? But ascorbate can inhibit bursting in cell types that depend on T-type channels for this kind of activity.
Depending on how you add your DA to the recording chamber, you can also keep it at low pH. We typically use a Razel pump to dilute a concentrated DA stock into the aCSF. For example, if you perfuse at 2ml/min, then you can deliver 20ul/min (e.g. 100 fold dilution). Make a syringe of DA in low (2-3) pH solution at 100x your desired bath concentration and introduce to the flowing aCSF near the inlet (you can use a Warner Instruments manifold to accomplish this).
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