Dopamine and Glutamate Receptor Assay

The assays of dopamine and glutamate receptors are an essential way to study the signaling to and functional operation of neural networks. With our extensive project experience, Creative Biolabs' most specialized neuroscience team could provide the most suitable solution for dopamine and glutamate receptor assays.

Overview of Dopamine and Glutamate Receptor

Dopamine receptors (DRs) are a family of G protein-coupled receptors consisting of seven transmembrane regions in the vertebrate central nervous system (CNS). There are five identified DRs that can activate different effectors by G-protein coupling and can also interact with other proteins to activate signals. Among them, D1R and D5R are D1-like receptors, which elevate intracellular cAMP levels upon activation, and D2R, D3R and D4R are D2-like receptors, which decrease intracellular cAMP levels upon activation. By binding to the major endogenous ligand dopamine, dopamine receptors regulate endocrine signaling to participate in a variety of neurological processes, including mood, cognition, learning, memory, and motor control.

Glutamate receptors are located in the cerebral cortex, hippocampus, cerebellum and striatum and play an influential role in learning, memory, neuronal plasticity and brain development through recognition with excitatory neurotransmitters. There are two types of glutamate receptors, ionotropic glutamate receptors that can couple with ion channels to alter membrane potential to mediate quick signaling, including N-methyl-D-aspartate receptor (NMDAR), kainic acid receptor (KAR) and α-amino-3-hydroxy-5 methyl-4 isoxazole receptor (AMPAR). Another class of metabotropic glutamate receptors are coupled to intramembrane G-proteins to alter slow cellular biochemical responses through effector enzymes and messenger signaling systems, thereby mediating a variety of cellular functions.

Fig.1 Proposed multimeric protein complex existing at the postsynaptic density. (Ciruela, 2005)

Dopamine and Glutamate Receptor Assay

The determination of dopamine receptors and glutamate receptors, as important receptors involved in neural signal transduction, is essential for the study of a variety of neurologically and psychologically relevant brain disorders such as schizophrenia, depression and Parkinson's disease. After agonist binding to D1-like receptors and activation of Gs proteins involved in the G-protein signaling pathway, in turn, the fluorescence intensity of the cAMP biosensor is increased by measuring the adenylate cyclase stimulation triggered by the cAMP-mediated cellular response. In this way, fluorescence emission in the enzyme-labeled instrument quantifies D1R and D5R activity in a dose-responsive manner.

D2R and D4R activity was achieved by measuring the fluorescence intensity after adenylate cyclase inhibition triggered by a cAMP-mediated cellular response involving activation of the D2R receptor on Gi/o proteins. In addition, the process of intracellular redistribution following D3R activation can also be detected. Similarly, glutamate receptor analysis is performed by activating the receptor and associated signaling pathways after in turn being treated with antagonists and using the associated fluorescence indication intensity changes as an indicator of the response to glutamate receptor activity. In addition, ELISA and WB are common methods available to perform receptor level analysis.

Fig.2 Levels of NMDA receptor subunits (GluN2A and GluN2B). (Wojtas, 2022)

Dopamine receptors and glutamate receptors, as key nodes connecting the entire neural signaling network, are closely linked to the maintenance of neurological function and the development of abnormal neurological disorders. Creative Biolabs has always been dedicated to neurologically relevant biochemical and pharmacological research and is able to provide our clients with high-quality dopamine and glutamate receptor assay services. Please feel free to contact us.

References

1. Ciruela, F.; et al. Heptaspanning membrane receptors and cytoskeletal/scaffolding proteins: focus on adenosine, dopamine, and metabotropic glutamate receptor function. J Mol Neurosci. 2005, 26(2-3): 277-92.
2. Wojtas, A.; et al. Effect of psilocybin and ketamine on brain neurotransmitters, glutamate receptors, DNA and rat behavior. Int J Mol Sci. 2022, 23(12): 6713.