Allosteric modulation of GluN1/GluN3 NMDA receptors by GluN1-selective competitive antagonists.

NMDA-type ionotropic glutamate receptors are critical for normal brain function and are implicated in central nervous system disorders. Structure and function of NMDA receptors composed of GluN1 and GluN3 subunits are less understood compared to those composed of GluN1 and GluN2 subunits. GluN1/3 receptors display unusual activation properties in which binding of glycine to GluN1 elicits strong desensitization, while glycine binding to GluN3 alone is sufficient for activation. Here, we explore mechanisms by which GluN1-selective competitive antagonists, CGP-78608 and L-689,560, potentiate GluN1/3A and GluN1/3B receptors by preventing glycine binding to GluN1. We show that both CGP-78608 and L-689,560 prevent desensitization of GluN1/3 receptors, but CGP-78608-bound receptors display higher glycine potency and efficacy at GluN3 subunits compared to L-689,560-bound receptors. Furthermore, we demonstrate that L-689,560 is a potent antagonist of GluN1FA+TL/3A receptors, which are mutated to abolish glycine binding to GluN1, and that this inhibition is mediated by a non-competitive mechanism involving binding to the mutated GluN1 agonist binding domain (ABD) to negatively modulate glycine potency at GluN3A. Molecular dynamics simulations reveal that CGP-78608 and L-689,560 binding or mutations in the GluN1 glycine binding site promote distinct conformations of the GluN1 ABD, suggesting that the GluN1 ABD conformation influences agonist potency and efficacy at GluN3 subunits. These results uncover the mechanism that enables activation of native GluN1/3A receptors by application of glycine in the presence of CGP-78608, but not L-689,560, and demonstrate strong intra-subunit allosteric interactions in GluN1/3 receptors that may be relevant to neuronal signaling in brain function and disease.

Adverse environmental impact: 30-year search for a definition.

Since passage of the Clean Water Act in 1972, there has been a long, unresolved struggle to define a key phrase in Section 316(b) of the act: "adverse environmental impact" (AEI). Section 316(b) requires that the best technology available be used in cooling-water intake structures to minimize AEI due to entrainment and impingement of aquatic organisms. Various attempts were made to evaluate and define AEI, including focused national conferences on impact assessment. Unresolved arguments regarding AEI were reinvigorated following the 1995 Consent Decree requiring EPA to propose new rules to implement Section 316(b). This article reviews and compares eight proposed definitions of AEI. Six of the definitions define AEI as impact expressed at the population or higher level of biological organization. The two remaining definitions are unrelated to populations: a 1% cropping of the near-field organisms and "one fish equals AEI". The latter definition is based on the desire of some stakeholders to define AEI as the loss of any public trust resources. Equating loss of public trust resources with AEI hampers consensus on a definition because a societal-based policy concept (public trust resources) is commingled with science-based definitions based on population effects. We recommend that a population-based definition of AEI be incorporated into Section 316(b) guidance and observe that this will not preclude a state from exercising its law and policy to protect public trust resources.