Endothelins negatively regulate glial glutamate transporter expression

Glutamate is the main excitatory neurotransmitter in the mammalian central nervous system which at high extracellular levels leads to neuronal over-stimulation and subsequent excitotoxic neuronal cell death. Both the termination of glutamatergic neurotransmission and the prevention of neurotoxic extracellular glutamate concentrations are predominantly achieved by the uptake of extracellular glutamate into astroglia through the high-affinity glutamate transporters, excitatory amino acid transporter-2/glutamate transporter-1 (EAAT-2/GLT-1) and EAAT-1/glutamate aspartate transporter (GLAST). Although several injury-induced growth factors such as epidermal growth factor (EGF) and transforming growth factor alpha (TGFalpha) potently stimulate the expression of glutamate transporters in cultured astroglia, GLT-1 and/or GLAST expression temporarily decreases during acute brain injuries eventually contributing to secondary neuronal cell death. We now demonstrate that the stimulatory influences of these injury-regulated growth factors are overridden by endothelins (ETs), a family of peptides also upregulated in the injured brain. Exposure of cultured cortical astroglia to ET-1, ET-2, and ET-3 resulted in a major loss of basal glutamate transporter expression after 72 hours and the complete prevention of the known stimulatory influences of dibutyryl cyclic (dbc)AMP, pituitary adenylate cyclase-activating polypeptide (PACAP), EGF, and TGFalpha on both GLT-1 and GLAST expression. With all ET isoforms, the inhibitory effects were detectable with similar low nanomolar concentrations and persisted in endothelin B-receptor deficient astroglia, suggesting that the inhibitory action is equally induced by endothelin A and B receptors. In astroglial cultures maintained with endothelins alone or in combination with PACAP, the inhibitory action was remarkably long-lasting and was still detectable after 7 days. In apparent contrast, glutamate transporter expression partially recovered between days 5 and 7 in cultures maintained with a combination of ETs and the injury-regulated growth factors EGF or TGFalpha. These findings point to ETs as major mediators of injury-dependent down-regulation of glial glutamate transporters and subsequent glutamate-induced brain damage.