L-TRANS-PYRROLIDINE-2,4-DICARBOXYLATE AND CIS-1-AMINOCYCLOBUTANE-1,3-DICARBOXYLATE BEHAVE AS TRANSPORTABLE, COMPETITIVE INHIBITORS OF THE HIGH-AFFINITY GLUTAMATE TRANSPORTERS

The ability of two conformationally restricted analogues of L-glutamat e to function as nontransportable inhibitors of plasma membrane L-glut amate transport was investigated in primary cultures of cerebellar gra nule cells and cortical astrocytes. L-trans-Pyrrolidine-2,4-dicarboxyl ic acid (L-trans-PDC) and cis-1-aminocyclobutane-1,3-dicarboxylic acid (cis-ACBD) behaved as linear competitive inhibitors of the uptake of D-[H-3]aspartate (used as a non-metabolizable analogue of L-glutamate) exhibiting K-i values between 40 and 145 mu M; L-trans-PDC being the more potent inhibitor in each preparation. However, both L-trans-PDC a nd cis-ACBD, over a concentration range of 1 mu M-5 mM, dose-dependent ly stimulated the release of exogenously supplied D-[H-3]aspartate fro m granule cells maintained in a continuous superfusion system. The sti mulated release was independent of extracellular calcium ions; essenti ally superimposable dose-response profiles being obtained in the absen ce and presence of 1.3 mM CaCl2 and yielding EC(50) values of 16-25 mu M and 18-220 mu M for L-trans-PDC and cis-ACBD, respectively. Stimula ted release of D-[H-3]aspartate was unaffected by either 300 mu M D-(- )-2-amino-5-phosphonopentanoic acid [D-APV; a selective antagonist of the N-methyl-D-aspartate (NMDA) receptor] or by 25 mu M 6-cyano-7-nitr oquinoxaline-2,3-dione [CNQX; a selective antagonist of the pha-amino- 3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) receptor]. The re lease of D-[H-3]aspartate following stimulation by either L-trans-PDC or cis-ACBD was however markedly attenuated following substitution in the superfusion medium of sodium ions by choline ions. Taken together, these results support an action of L-trans-PDC and cis-ACBD consisten t with that of being competitive substrates rather than non-transporta ble blockers of the plasma membrane L-glutamate uptake system.