Dopaminergic neuron markers
A summary of the best dopaminergic neuron markers for your research.
Dopaminergic neurons produce dopamine, a monoamine neurotransmitter with roles in neurological functions such as mood and reward, as well as diverse physiological functions across different brain regions. Midbrain dopaminergic neurons, also referred to as midbrain dopamine neurons, are key neuron populations located in specific brain regions such as the substantia nigra pars compacta (SNc) and ventral tegmental area (VTA).
The progressive loss of dopaminergic neurons is the cause of many of the motor symptoms associated with Parkinson’s disease. Specific neuron subtypes within the midbrain are particularly vulnerable in Parkinson's disease, highlighting the importance of understanding their molecular and functional diversity. A widely used treatment for Parkinson’s disease is Levodopa, which is a pure form of the precursor to dopamine, L-DOPA.
There are several well-characterized dopamine markers that can aid your research. Gene expression profiling is commonly used to distinguish different neuron subtypes within midbrain dopaminergic neurons.
Tyrosine hydroxylase
An enzyme that converts L-tyrosine to L-3,4-dihydroxyphenylalanine (L-DOPA), which is a dopamine precursor, tyrosine hydroxylase is classified as an aromatic amino acid hydroxylase1-4.
Figure 1. Anti-Tyrosine Hydroxylase antibody [EP1532Y] - staining Tyrosine Hydroxylase in mouse brain tissue sections by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections).
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Dopamine transporter (DAT)
A transmembrane transporter that controls the reuptake of extracellular dopamine into presynaptic neurons and pumps dopamine out of the synapse into the cytosol. DAT (also known as the dopamine active transporter) is vital in maintaining sufficient dopamine levels in the neuron for release. Alterations in dopamine transporter function have been implicated in neuropsychiatric disorders such as bipolar disorder and attention deficit hyperactivity disorder5-8.
Figure 2. Anti-Dopamine Transporter antibody [6V-23-23] staining dopamine transporter in mouse brain (substantia nigra) tissue sections by immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).
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GIRK2
A G-protein regulated potassium channel expressed within certain dopaminergic neurons of the substantia nigra, particularly enriched in neurons of the ventral tier of the substantia nigra9-13.
Figure 3. Mouse substantia nigra tissue sections stained with anti-GIRK2 (ab65096).
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NURR1
A transcription factor that induces TH expression and subsequently dopaminergic neuron differentiation, NURR1 is one of several transcription factors involved in neuron differentiation, particularly in mda neuron differentiation14.
Figure 4. Rat cerebral nerve cells stained with anti-nurr1 (ab41917).
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LMX1b
A transcription factor involved in a number of processes during dopaminergic neuron development15. LMX1b is also used in protocols to differentiate stem cells, including pluripotent stem cells, into dopaminergic neurons for research and therapeutic applications.
Figure 5. Immunohistochemical analysis of paraffin-embedded Rat E14.5 embryo tissue labelling LMX1b with ab259926 at 1/2000 (0.278 μg/ml) dilution, followed by a ready-to-use LeicaDS9800 (Bond™ Polymer Refine Detection).
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Fluorescent false neurotransmitters (FFNs)
Fluorescent false neurotransmitters (FFNs) are probes that act as novel optical tracers, allowing imaging of neurotransmitter release from individual presynaptic terminals in the brain. They are designed to loosely mimic the overall topology and physical properties of monoamine neurotransmitters and have been engineered to have fluorescence properties.
FFNs have the capability for increasing the understanding of both fundamental and applied neurobiological research, including research into neurodegeneration and drug addiction.
Advantages of using fluorescent false neurotransmitters:
- Optically study various aspects of synaptic transmission
- Compatible with GFP tags, including Alexa Fluor 488
- Photostable
- Suitable for standard fluorescent microscopy and two-photon fluorescence microscopy
- Allow imaging of neurotransmitter release at single-cell resolution
Applications for FFN102, FFN202, and FFN511:
- Measure localization and activity of dopaminergic presynaptic terminals.
- Measure pH of secretory vesicles
- Visualize dopamine release from individual presynaptic terminals
FFN imaging can also be combined with in situ hybridization to correlate neurotransmitter release with gene expression in individual neurons.
FFN102 (Mini102)
- Rodent DAT and VMAT2 substrate. Enables two-photon microscopic imaging of localization and activity of dopaminergic presynaptic terminals in the striatum of mouse acute brain slice.
- Allows analysis of dopamine release at the single-cell level.
- More selective for dopaminergic synapses than FFN511.
- Exhibits greater fluorescence emission in neutral than acidic environments allowing optical measurement of synaptic vesicle content release.
Figure 6. FFN102 molecular structure.
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FFN102 (Mini 102), Fluorescent DAT and VMAT2 substrate
FFN202 (Mini202)
- Rodent VMAT1 and VMAT2 substrate.
- Enables in situ pH measurement of secretory catecholamine vesicles in PC-12 cells.
Figure 7. FFN202 molecular structure.
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FFN202 (Mini 202), Fluorescent VMAT1 and VMAT2 substrate
FFN511
- A fluorescent probe for optical imaging and measurement of synaptic activity in the brain.
- Substrate for the synaptic vesicle monoamine transporter (VMAT2).
- Can be used to visualize labeled cell bodies in dopaminergic neuron populations.
- FFN511 fluoresces sufficiently to provide resolution at the individual synaptic level but at concentrations that do not interfere with normal synaptic function. Inhibits 5HT binding to VMAT2 with an IC50 of 1 μM (comparable to dopamine itself).
Figure 8. FFN511 molecular structure
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FFN511, Fluorescent substrate for VMAT2
References
1. Hartfield, E. M.,, Yamasaki-Mann, M.,, et al. Physiological characterisation of human iPS-derived dopaminergic neurons. PLoS One 9 , (2014)
2. Luo , F. C., Wang, S. D., et al. Protective effect of panaxatriol saponins extracted from Panax notoginseng against MPTP-induced neurotoxicity in vivo. J. Ethnopharmacol. 133 ,448-453 (2011)
3. Ness, D. K., Foley, G. L., et al. Effects of 3-iodo-L-tyrosine, a tyrosine hydroxylase inhibitor, on eye pigmentation and biogenic amines in the planarian, Dugesia dorotocephala. Fundam. Appl. Toxicol. 30 ,153-161 (1996)
4. Smythe, G. A., Bradshaw, J. E. Different acute effects of the tyrosine hydroxylase inhibitors alpha-methyl-p-tyrosine and 3-iodo-L-tyrosine on hypothalamic noradrenaline activity and adrenocorticotrophin release in the rat. Aust. J. Biol. Sci. 36 ,519-523 (1983)
5. Austin, L. S., Lydiard, B, et al. Dopamine blocking activity of clomipramine in patients with obsessive-compulsive disorder. Biol. Psychiatry 30 ,225-232 (1991)
6. Rothman, R. B., , Baumann, M. H., et al. Dopamine transport inhibitors based on GBR12909 and benztropine as potential medications to treat cocaine addiction. Biochem. Pharmacol. 75 ,2-16 (2008)
7. Stahl, S. M., Pradko, J. F., et al. Review of the Neuropharmacology of Bupropion, a Dual Norepinephrine and Dopamine Reuptake Inhibitor. Prim. Care Companion J. Clin. Psychiatry 6 ,159-166 (2004)
8. Tanda, G., Li, S. M., et al. Relations between stimulation of mesolimbic dopamine and place conditioning in rats produced by cocaine or drugs that are tolerant to dopamine transporter conformational change. Psychopharmacology (Berl). 229 ,307-321 (2013)
9. Hartfield, E. M., Yamasaki-Mann, M., et al. Physiological characterisation of human iPS-derived dopaminergic neurons. PLoS One 9 , (2014)
10. Kaufmann, K., Romaine, I., et al. ML297 (VU0456810), the first potent and selective activator of the GIRK potassium channel, displays antiepileptic properties in mice. ACS Chem. Neurosci. 4 ,1278-1286 (2013)
11. Kobayashi, T., Ikeda, K.,, Kumanishi, T. Inhibition by various antipsychotic drugs of the G-protein-activated inwardly rectifying K(+) (GIRK) channels expressed in xenopus oocytes. Br. J. Pharmacol. 129 ,1716-1722 (2000)
12. Kobayashi, T., Washiyama, K., Ikeda, K. Inhibition of G protein-activated inwardly rectifying K+ channels by ifenprodil. Neuropsychopharmacology 31 ,516-524 (2006)
13. Reyes, S., Fu, Y., et al. GIRK2 expression in dopamine neurons of the substantia nigra and ventral tegmental area. J. Comp. Neurol. 520 ,2591-2607 (2012)
14. Jankovic J., Chen, S., , Le, W. D. The role of Nurr1 in the development of dopaminergic neurons and Parkinson’s disease. Prog. Neurobiol. 77 ,128-138 (2005)
15. Nakatani, T., , Kumai, M.,, et al. Lmx1a and Lmx1b cooperate with Foxa2 to coordinate the specification of dopaminergic neurons and control of floor plate cell differentiation in the developing mesencephalon. Dev. Biol. 339 ,101-113 (2010)