Extensive research has focused on the neurotransmitter dopamine because of its importance in the mechanism of action of drugs of abuse (e.g. cocaine and amphetamine), the role it plays in psychiatric illnesses (e.g. schizophrenia and Attention Deficit Hyperactivity Disorder), and its involvement in degenerative disorders like Parkinson's and Huntington's disease. Under normal physiological conditions, dopamine is known to regulate locomotor activity, cognition, learning, emotional affect, and neuroendocrine hormone secretion. One of the largest densities of dopamine neurons is within the striatum, which can be divided in two distinct neuroanatomical regions known as the nucleus accumbens and the caudate-putamen. The objective is to illustrate a general protocol for slice fast-scan cyclic voltammetry (FSCV) within the mouse striatum. FSCV is a well-defined electrochemical technique providing the opportunity to measure dopamine release and uptake in real time in discrete brain regions. Carbon fiber microelectrodes (diameter of~7 μm) are used in FSCV to detect dopamine oxidation. The analytical advantage of using FSCV to detect dopamine is its enhanced temporal resolution of 100 milliseconds and spatial resolution of less than ten microns, providing complementary information to in vivo microdialysis.
Video LinkThe • There are numerous carbon fiber microelectrodes fabrication methods since most are made in-house. Typically what dictates the electrode fabrication details is the electrochemical technique that is applied to the electrode (e.g. amperometry vs. FSCV). For FSCV, microelectrodes can be made in-house using the following three-step procedure. For a more complete description of carbon fiber electrode fabrication, see a recent JOVE article 1 . However, note that the electrodes described below are cylindrical carbon fiber microelectrodes, which require fewer steps to fabricate versus the amperometric carbon fiber microelectrodes from the above-mentioned protocol. This simplified protocol does not require boiling the carbon fiber in acetone, fire-polishing the glass capillaries, or using epoxy to seal the glass-fiber junction.• Using vacuum suction aspirate a carbon fiber (diameter 7 μm; Goodfellow Oakdale, PA) into a borosilicate glass capillary with microfilament (length 10 cm, o.d. 1.2 mm, i.d. 0.68 mm; A-M systems, Carlsborg, WA).• Place the threaded capillary into the electrode puller (Narishige, Tokyo, Japan) where the capillary is pulled in half. Output settings for the electrode puller do vary from lab to lab. For reference, our output settings for the puller are 90.7 main magnet, 23.2 sub-magnet, and 53.4 for the heater. The output settings should be empirically refined to generate a glass taper that is approximately 4.4 mm in length, with a tight seal around the carbon fiber.• Under a microscope (Olympus, Tokyo, Japan), trim the carbon fiber (using a scalpel blade) extending from the glass tip allowing approximately 50-200 μm of the carbon fiber to protrude from the tightly sealed interface.• S...