Cyclic Voltametry for the

Detection of Dopamine in vivo

 Dopamine
• Neurotransmitter
– small molecule chemical messenger
• Important for motor and
cognitive functions
– Deficts in dopamine levels cause
Parkinson Disease
• Regulates reward Structure of Dopamine.
4-(2-aminoethyl)
– Dopamine increases after drugs of benzene-1,2-diol
abuse like cocaine
 Dopaminergic Neurons
• Dopamine is synthesized in dopaminergic neurons
and packaged into membrane bound vesicles
• Electrical action potential initiates the release of
dopamine
• Dopamine vesicles undergo exocytosis
– Spills out into the extracellular space
– Can be detected

The dompaminergic neuron can

release dopamine into the
extracellular space where its
contents can be detected by
target neurons.
 Detection of Dopamine
• Exocytosis of dopamine from vesicle occurs on
a millisecond time scale
• Sensor must be fast, sensitive, and selective
since dopamine concentrations are low
• Fast scan cyclic voltametry is the dominant
electrochemical technique used
Dopamine Dopamine-o-quinone
HO NH2 - O NH2
-2e
+ 2 H+
HO O
 Carbon Fiber Microelectrodes
• Carbon Fiber
Microelectrodes (CFME)
are electrodes in which a
carbon fibers serve as the
electroactive area
• Advantages:
– Carbon fibers are biological
compatible to cells
– Small size (less than 10um
in diameters) allows for Carbon Fiber Microelectrode
implantation in vivo
• Commonly used with cyclic
voltametry
 Voltammetry
• Electrochemical technique in which the
current (I) is measured as a function of voltage
(Eapp).
• There are several kinds:
– Linear Scan voltametry (Polarography)
– Differential pulse polarography
– Square-wave voltametry
– Cyclic Voltametry
 Fast-scan Cyclic Voltametry
1.3 V
400 V/s

-0.4 V
8.5 ms 100 ms

Scan electrode from a holding potential to a switching potential and back at a

high scan rate. Repeat these scans every 100 ms.

Dopamine Dopamine-o-quinone
HO NH2 - O NH2
-2e
+ 2 H+
HO O

As potential is ramped up, dopamine is oxidized to dopamine-o-quinone.

As potential is ramped down, dopamine-o-quinone reduced back to dopamine.
 Fast-Scan Cyclic Voltametry
Cyclic voltammograms Background-subtracted
No dopamine
Dopamine present
cyclic voltammogram of dopamine
oxidation

reduction

Fast scan rate cause a large background The background current is stable and can
charging current due to charging be subtracted out to obtain a background-
the double layer. subtracted cyclic voltammogram for dopamine.

When dopamine is added (red line), The position of the peaks help identify the
the differences are small. molecule being detected.
 Fast-scan cyclic voltametry
10ss
10
-0.4VV
-0.4
dopamine
reduction 33
Ag/AgCl
E EvsvsAg/AgCl

00
1.0VV
1.0 i, nA

-5
-5
dopamine i, nA
oxidation
-0.4VV
-0.4
dopamine present

•Data can also be depicted as a color plot to show many CVs over time.
•Current is in color and shows when dopamine is present.
 Results
Electrically-Stimulated release in a rat

•Carbon-fiber microelectrode implanted in nucleus accumbens

•Nucleus accumbens regulates reward
•Electrically stimulate cell bodies in ventral tegmental area
•Size of dopamine evoked depends on frequency of stimulation
• Color plots look like dopamine
 Results
Spontaneous dopamine transients in a rat

Spontaneous dopamine transients in a rat after cocaine

•Cocaine increases the concentration and the length of time for dopamine
signaling.
 Advantages
• Allows real-time detection of dopamine in
behaving animals
• Can determine during what behaviors
dopamine is released
• Can determine how drugs affect dopamine
signaling in the brain