High Performance Liquid Chromatography of Neurotransmitters in Microdissected Brains of Inebriated Zebrafish

PAC Gym

Neurotransmitters work with the brain to influence and regulate a plethora of processes and behavior. Zebrafishes have become a popular model to quantify changes in behavior caused by changes in neurotransmitter levels because they are vertebrates with a fully sequenced genome that are easier to nurture than their mammalian peers. We seek to investigate alcohol’s effect on two wild-type zebrafish, AB and TU, by measuring their biogenic amine and metabolite levels. For 21 consecutive days, half of the AB and TU zebrafish were subjected to 1-hour long ethanol exposures in a tank containing 0.125% (v/v) ethanol, while the other half were placed in control tank containing system water only. After the ethanol exposures, the zebrafish were sacrificed, and their brains dissected. During dissection, half of the zebrafish from each of the two groups were randomly selected to have their fore and mid/hind brains microdissected. The changes in all neurotransmitter levels were then quantified for the whole, fore, and mid/hind brain samples using high performance liquid chromatography with electrochemical detection using an optimized phosphate buffer with a C-18 reverse-phase column. These changing conditions offer exciting methods to study how neurotransmitter levels interact with one another while under the effects of drugs of abuse.