Extended Data Fig. 7: Deletion of A2B receptors in hippocampal astrocytes.
From: Adenosine signalling to astrocytes coordinates brain metabolism and function
(a) Representative immunofluorescence images showing transgene expression after the microinjections of AAV5-Gfap-eGFP-iCre vector into the hippocampus in mice. Summary data illustrate quantification of brain cells expressing GFP and identified by immunohistochemical labeling to also express either glial fibrillary acidic protein (GFAP), Ionized calcium binding adaptor molecule 1 (Iba1), neuronal nuclear antigen (NeuN), or myelin basic protein (MBP) (n = 3 mice). Gfap promoter displayed high selectivity for astrocytes. Scale bars = 50 µm; (b) Hippocampal astrocytes of Adora2bflox/flox mice were transduced to express iCre recombinase (microinjections of AAV5-Gfap-eGFP-iCre vector) or control transgene (microinjections of AAV5-Gfap-eGFP vector), followed by isolation of transduced astrocytes by FACS. Gating strategy used for isolation of transduced cells: hippocampal astrocyte population was first gated based on morphology by size and complexity (FSC and SSC) and then gated based on the expression of GFP (control/GFP+: Adora2bflox/flox mice injected with AAV5-Gfap-eGFP; A2BR-Astro-KD/GFP+: Adora2bflox/flox mice injected with AAV5-Gfap-eGFP-iCre; brain tissue of wild-type naïve mice was used as a negative control [wild-type/GFP-]). Expression of iCre recombinase in hippocampal astrocytes of Adora2bflox/flox mice reduced A2B receptor expression. Data are presented as individual values and means ± s.e.m. Numbers in parentheses indicate the numbers of animals per experimental group. P values, one-tailed Student’s t-test.
