Fig 1: Adult mouse hippocampal astrocytes express GluN2C-NMDA receptors.(A) Top: patch RT-PCR strategy. Bottom: representative voltage responses to current steps in CA1 pyramidal neuron (far left, black), and current responses to voltage steps in astrocytes in stratum radiatum (SR, green), stratum oriens (SO, yellow), and stratum lacunosum moleculare (SLM, blue). Scale bars: neuron, 40 mV, 500 ms; astrocytes, 6 nA, 200 ms. (B) Summary of I-V curves of astrocytes obtained before collecting RNA. N = 39 cells, 36 slices, 9 mice for each layer. (C) Summary of RT-PCR analysis for mRNA encoding indicated NMDAR subunits: Grin1 (1), Grin2a (2A), Grin2b (2B), and Grin2c (2C). Control samples (no patch) were obtained by inserting the electrode into the slice without patching cells. Three technical replicates were performed for each condition. (D) Western blots of co-immunoprecipitations performed using adult mouse hippocampal extracts with the indicated antibodies where anti-Cre antibody is used as a negative control. The blots are probed for GluN2A/2B (top row), GluN2C (middle row) or GluN1 (bottom row). (E) Representative brain sections of Grin2ciCre/+ mice crossed to a tdTomato reporter line (Ai9), showing hippocampal area CA1 immunolabelled for GFAP, tdTomato and NeuN; scale bars, 160 µm (top), 25 µm (bottom). (F) Quantification of % cells that are positive for tdTomato amongst the counts of NeuN-labelled cells in stratum pyramidale (left, total n = 3155 cells) and SR (middle, total n = 619 cells), and GFAP-labelled cells (right, total n = 3149 cells). N = 10 fields of view of area CA1 from individual hippocampal sections (n = 10) obtained from 5 mice. Data are presented as mean ± s.e.m.