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MeAIB is a competitive substrate with specificity for system A transport over other glutamine transporters (Christensen et al. 1965), and 20 mm MeAIB reduced IGln by 92.9 �� 2.8% (Fig. 1C; n= 10; P 0.05). We also tested the effects of the EAAT activator d-aspartate on IGln. Bath application of 200 ��m d-aspartate did not have a significant effect on IGln (Fig. 1C; 110.4 �� 5.8% of control; n= 3; P > 0.05), confirming that this current is not mediated by EAATs and demonstrating that d-aspartate does not http://www.selleckchem.com/products/lee011.html affect glutamine transport on the MNTB cell, which is an important control for later experiments. Astrocytes are known to express EAAT1 and EAAT2 transporters, which are responsible for removing glutamate from the synaptic cleft following neurotransmission (Bergles et al. 1999; Danbolt, 2001). To investigate EAAT-mediated glutamate transport on astrocytes in the MNTB, we whole-cell voltage-clamped astrocytes http://www.selleckchem.com/products/bmn-673.html and puff-applied d-aspartate, an EAAT substrate that does not activate ionotropic glutamate receptors. Astrocytes adjacent to the calyx of Held synapse could be identified by bathing slices in sulforhodamine 101 (Fig. 2A�CC; Nimmerjahn et al. 2004; Kafitz et al. 2008). Additionally, electrical membrane properties were used to confirm astrocytic recordings (Muller et al. 2009): the resting membrane potential was http://www.selleck.cn/products/AZD6244.html of 200 ��m glutamate (in the presence of glutamate receptor inhibitors APV, MK801 and NBQX) induced a current of �C31.3 �� 10.5 pA (n= 5), which was inhibited 86.7 �� 11.8% by addition of the EAAT inhibitor TBOA (n= 5; P