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EAAT3 - "High throughput SSM recordings of EAAT3 (SLC1A1) activity on the SURFE2R 96SE"

Icon 96SE   SURFE2R 96SE application note   logo pdf   (1.5 MB)

 Summary:

The excitatory amino acid transporter 3 (EAAT3; also known as EAAC1) is a sodium-dependent neuronal uptake transporter encoded by the SLC1A1 gene. It plays a major role in the reuptake of glutamate from the synaptic cleft, thereby maintaining a low extracellular concentration of glutamate and regulating excitatory neurotransmission. EAAT3 is also involved in the uptake of aspartate and cysteine into the cells. The transporter is highly expressed in mature neurons, where it is distributed in somata and dendrites.

EAAT3 functions as a co-transporter, coupling the uphill substrate transport into the cells to the electrochemical gradients of sodium and potassium. The stoichiometry of transport is 1 glutamate with 3 Na+ and 1 H+ moving into the cell to 1 K+ moving out of the cell. Therefore EAAT3 is an electrogenic transporter, generating a net charge flow.

Dysfunction of glutamate transporters leads to increased extracellular glutamate levels, thereby causing neurotoxicity and neurodegeneration. Regulatory mechanisms facilitating EAAT3 function are, therefore, interesting as targets for the treatment of neurodegenerative diseases.

Here we present EAAT3 activity measurements on the SURFE2R 96SE instrument using purified plasma membrane of CHO cells expressing EAAT3. The transporter was activated by glutamate and blocked by six different known inhibitors with IC50 values similar to those found in the literature. 

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