PET imaging of P2 purinoceptors

Purinergic receptors (purinoceptors) include the P1 purinoceptors (adenosine receptors), and P2 purinoceptors which are preferably activated by ATP and other substrates than adenosine. ATP and its breakdown product adenosine are primitive signalling molecules, that can modulate the postsynaptic response and presynaptic release of their cotransmitter. Modulatory functions of purinoceptors are observed in most nerves, including cholinergic transmission in motor nerves and neuromuscular junctions, sensory-motor nerves, sympathetic and parasympathetic nerves, and in the central nervous system (Burnstock and Verkhratsky, 2012).

ATP is used as signalling molecule also by other cells, including vascular endothelial cells, blood cells, muscle, liver, spleen. Renal P2 purinoceptors affect tubular function and renal perfusion. Cells that participate in ATP signalling have ATP containing vesicles, and the vesicular nucleotide transporter (SLC17A9), which concentrates ATP into the vesicles.

ATP is synthesized in mitochondria, and its concentration in cytosol is high, about 3-10 mM, and can be much higher in ACh and serotonin containing vesicles, and in lysosomes; in extracellular space [ATP] is in nM range. Some ATP can diffuse from cells through ion channels, for example mechanical stimulation may activate chloride channels. Cell damage leads to increased extracellular ATP, which can signal tissue damage to the inflammatory system. At the sites of inflammation, extracellular ATP concentrations can be 4-5 orders of magnitude higher than in healthy tissue (Di Virgilio et al., 2017). Extracellular nucleotides are rapidly catabolized by ectonucleases.

P2 purinoceptors include the metabotropic G protein coupled P2Y receptors, and ionotropic ligand-gated ion channels P2X.

P2Y receptors

P2Y receptors are stimulated by nucleotides such as ATP, ADP, UTP, UDP and UDP-glucose, with different substrate specificity. Several P2Y receptor subtypes have been identified. In humans, at least P2Y1, P2Y2, P2Y4, P2Y6, P2Y11, P2Y12, P2Y13, and P2Y14 have been found.

PET tracers for P2YRs are being developed, but not in use.

P2X receptors

ATP-gated nonselective cation channels (P2XRs) are trimers (either homomeric or heteromeric) of subunits (P2X1 - P2X7). Three ATP binding sites are probably located between the subunits.

P2X receptors are expressed by wide array of cell types. For example, P2X1R is expressed on astrocytes, oligodendrocytes, smooth, skeletal and heart muscle, osteoblasts and osteoclasts, adipocytes, platelets, endothelial cells, and exocrine secretary cells; P2X2R on neurons, astrocytes, oligodendrocytes, osteoblasts and osteoclasts, cartilage, keratinocytes, erythrocytes, and endocrine secretary cells; P2X3R on cardiac muscle, keratinocytes, and endothelial cells; P2X4R on neurons, microglial cells, cardiac muscle, osteoblasts and osteoclasts, endothelial cells, white blood cells, erythrocytes, and endo- and exocrine cells; P2X5R on neurons, astrocytes, osteoblasts, keratinocytes, and epithelial cells; P2X6R on neurons, heart muscle, epithelial cells, and endocrine secretory cells. Skeletal muscle cells and sympathetic neurons contain all P2X subtypes.

P2X7 receptor

P2X7R is expressed in many cell types, including sympathetic neurons, astrocytes, oligodendrocytes, microglial cells, probably also CNS neurons, skeletal muscle cells, osteoblasts and osteoclasts, keratinocytes, fibroblasts, epithelial cells, white blood cells, erythrocytes, and endo- and exocrine cells. In the brain, P2X7R levels are highest in microglia, and the receptor is important in regulation of immune responses, including the neuroinflammatory cascades that precede and promote many neurodegenerative brain diseases.

P2X7R is structurally and functionally distinct from the other P2X receptors. Intense stimulation of P2X7Rs results in the formation of a large transmembrane pore that is permeable to molecules of size up to 900 Da (Burnstock and Verkhratsky, 2012), further increasing the extracellular [ATP], and can lead to activation of caspases and cell death.

The gene of the P2X7R is polymorphic, including many splice variations and single nucleotide polymorphisms (Di Virgilio et al., 2017). These are associated with different diseases, including multiple sclerosis, bipolar disorder, and increased risk for bone fractures.

Various inflammatory mediators can upregulate P2X7R expression on macrophages and other cell types, while anti-inflammatory mediators can downregulate the expression (Bartlett et al., 2014).

Several PET tracers targeting P2X7R have been synthesized, but usually found not to be suitable for clinical studies, mainly because of poor BBB permeability (Fantoni et al., 2017). [11C]GSK1482160 (Territo et al., 2017; Han et al., 2017), [11C]JNJ-54173717 (Ory et al., 2016), and [11C]SMW139 are new promising P2X7R tracers for imaging neuroinflammation.

See also:


Burnstock G, Verkhratsky A: Purinergic Signalling and the Nervous System. Springer, 2012. doi: 10.1007/978-3-642-28863-0.

Coddou C, Yan Z, Obsil T, Huidobro-Toro JP, Stojilkovic SS. Activation and regulation of purinergic P2X receptor channels. Pharmacol Rev. 2011; 63: 641-683.

Di Virgilio F, Dal Ben D, Sarti AC, Giuliani AL, Falzoni S. The P2X7 receptor in infection and inflammation. Immunity 2017; 47(1): 15-31.

Erb L, Weisman GA. Coupling of P2Y receptors to G proteins and other signaling pathways. WIREs Membr Transp Signal. 2012; 1: 789-803.

Hattori M, Gouaux E. Molecular mechanism of ATP binding and ion channel activation in P2X receptors. Nature 2012; 485: 207-213.

Ho HTB, Wang J. (2014): The Nucleoside Transporters CNTs and ENTs. In: Drug Transporters: Molecular Characterization and Role in Drug Disposition, 2nd ed (eds You G, Morris ME), John Wiley & Sons, Inc., Hoboken, NJ. doi: 10.1002/9781118705308.ch7.

Jacobson KA, Linden J (eds): Pharmacology of Purine and Pyrimidine Receptors. Academic Press, 2011. ISBN: 978-0-12-385526-8

Menzies RI, Tam FW, Unwin RJ, Bailey MA. Purinergic signaling in kidney disease. Kidney Int. 2017; 91: 315-323. doi: 10.1016/j.kint.2016.08.029.

Zimmermann H. Extracellular ATP and other nucleotides - ubiquitous triggers of intracellular messenger release. Purinergic Signal. 2016; 12: 25-57.

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Created at: 2016-05-12
Updated at: 2018-07-12
Written by: Vesa Oikonen