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Neuronal inhibition is mediated by glycine and/or GABA. Inferior colliculus (IC) neurons receive glycinergic and GABAergic
inputs, whereas inhibition in hippocampus (HC) predominantly relies on GABA. Astrocytes heterogeneously
express neurotransmitter transporters and are expected to adapt to the local requirements regarding neurotransmitter
homeostasis. Here we analyzed the expression of inhibitory neurotransmitter transporters in IC and HC astrocytes using
whole-cell patch-clamp and single-cell reverse transcription-PCR. We show that most astrocytes in both regions expressed
functional glycine transporters (GlyTs). Activation of these transporters resulted in an inward current (IGly) that
was sensitive to the competitive GlyT1 agonist sarcosine. Astrocytes exhibited transcripts for GlyT1 but not for
GlyT2. Glycine did not alter the membrane resistance (RM) arguing for the absence of functional glycine receptors (GlyRs).
Thus, IGly was mainly mediated by GlyT1. Similarly, we found expression of functional GABA transporters (GATs) in all IC
astrocytes and about half of the HC astrocytes. These transporters mediated an inward current (IGABA) that was sensitive to
the competitive GAT-1 and GAT-3 antagonists NO711 and SNAP5114, respectively. Accordingly, transcripts for GAT-1 and
GAT-3 were found but not for GAT-2 and BGT-1. Only in hippocampal astrocytes, GABA transiently reduced
RM demonstrating the presence of GABAA receptors (GABAARs). However, IGABA was mainly not contaminated
by GABAAR-mediated currents as RM changes vanished shortly after GABA application. In both regions, IGABA
was stronger than IGly. Furthermore, in HC the IGABA/IGly ratio was larger compared to IC. Taken together, our
results demonstrate that astrocytes are heterogeneous across and within distinct brain areas. Furthermore, we
could show that the capacity for glycine and GABA uptake varies between both brain regions.
Anisotropy of tracer-coupled networks is a hallmark in many brain regions. In the past, the topography of these networks was analyzed using various approaches, which focused on different aspects, e.g., position, tracer signal, or direction of coupled cells. Here, we developed a vector-based method to analyze the extent and preferential direction of tracer spreading. As a model region, we chose the lateral superior olive—a nucleus that exhibits specialized network topography. In acute slices, sulforhodamine 101-positive astrocytes were patch-clamped and dialyzed with the GJ-permeable tracer neurobiotin, which was subsequently labeled with avidin alexa fluor 488. A predetermined threshold was used to differentiate between tracer-coupled and tracer-uncoupled cells. Tracer extent was calculated from the vector means of tracer-coupled cells in four 90° sectors. We then computed the preferential direction using a rotating coordinate system and post hoc fitting of these results with a sinusoidal function. The new method allows for an objective analysis of tracer spreading that provides information about shape and orientation of GJ networks. We expect this approach to become a vital tool for the analysis of coupling anisotropy in many brain regions