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. 2024 Apr 30;15:3661. doi: 10.1038/s41467-024-47964-4

Fig. 2. Activation of astrocytes or neurons in the NAc increases extracellular adenosine levels.

Fig. 2

a Schematic of microinjection of AAVs expressing neuronal Ado1.0 and astrocytic hM3Dq DREADD and optic fiber placement in the NAc of WT mice, illustrated by Sara Kobayashi. b Typical examples of EEG, EMG, and adenosine signals for 1 h in the NAc of a WT mouse. The EMG trace is shown as root mean square and the red line indicates level 5 activity. c Mean NAc adenosine levels before and after each state transition. Data [Transitions examined over 5 independent experiments: Wake-SWS, n = 30 (Ado1.0) or 26 (Ado1.0mut); SWS-Wake, n = 30 (Ado1.0) or 25 (Ado1.0mut); SWS-REMS, n = 30 (Ado1.0 or Ado1.0mut); REMS-SWS, n = 27 (Ado1.0) or 26 (Ado1.0mut)] are presented as mean ± SEM (shaded areas). Time course (d) and mean (e) adenosine levels after chemogenetic stimulation of astrocytes in the NAc of WT. Data (n = 5 biologically independent animals/group) are presented as mean ± SEM [shaded areas in (d) or error bars in (e)]. d Horizontal bars indicate false discovery rate Q. e Unpaired 2-tailed t-test compared with vehicle (saline) injection. f Schematic of microinjection of AAVs expressing neuronal Ado1.0 and hM3Dq DREADD and optic fiber placement in the NAc of WT mice, illustrated by Sara Kobayashi. Time course (g) and mean (h) adenosine levels after chemogenetic stimulation of neurons in the NAc of WT. Data [g, h, n = 7 (vehicle) and n = 8 (CNO) biologically independent animals in each group] are presented as mean ± SEM [shaded areas in (g) or error bars in (h)]. g Horizontal bar indicates false discovery rate Q. h Unpaired 2-tailed t-test compared with vehicle (saline) injection. Source data have been deposited in the Figshare database [10.6084/m9.figshare.25468084]. Abbreviations: AAV adeno-associated virus, CNO clozapine N-oxide, DREADD designer receptors exclusively activated by designer drugs, EEG electroencephalogram, EMG electromyogram, GFAP glial fibrillary acidic protein, GRABAdo1.0/mut G protein-coupled receptor-activation-based adenosine sensor Ado1.0 or non-binding mutant Ado1.0mut, hSyn human synapsin, NAc nucleus accumbens, pA polyadenylation signal, REMS rapid eye movement sleep, SEM standard error of the mean, SWS slow-wave sleep, WPRE woodchuck hepatitis virus posttranscriptional regulatory element, WT wild type.