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. Author manuscript; available in PMC: 2020 Jul 1.

Published in final edited form as: Exp Neurol. 2019 Feb 26;317:66–77. doi: 10.1016/j.expneurol.2019.01.014

Figure 1. — The brains of neonatal mice and preterm infants expressed A₁R receptors. (A) Immunostaining results showed the specificity of A₁R antibody but not A_2AR antibody. Using the tested A₁R antibody, A₁R immunostaining showed a punctate distribution of this receptor in the cortex of wild type adult mice, but not in that of A₁R deficient mice (25.33±2.4 and 8.67±0.88 puncta, respectively) (top). In contrast, several antibodies against A_2AR showed non-specific immunoreactivity in the nucleus and the cytoplasm in both wild type and A_2AR deficient mice (bottom). No punctate pattern was observed (6±1.53 puncta for WT and 4.33±0.3 puncta for A_2AR knockout). Representative images are shown here. The quantitative results are shown on the right. Blue arrows indicate puncta. Scale bar, 10 μm. (B) The punctate pattern of A₁R could be detected in the cortex of P3 pups. The A₁R puncta appeared more discrete at a lower magnification as the animals mature (10 day-old and 1 month-old pups). (C) A₁R distributed in a punctate pattern in medulla, cerebellum, thalamus and hippocampus. The puncta were more prominent in medulla and cerebellum. Scale bar, 20 μm. (D) Immunostaining showed a punctate pattern of A₁R in the cortex of preterm infants at the corrected age of 26–27 weeks. Representative images of the cortex from 6 preterm brains are shown. Scale bar, 10 μm. (E) A₁R colocalized with a synaptic protein, PSD-95, in frozen sections of neonatal mouse brain (upper panel) and paraffin sections of preterm infant brains (lower panel). Scale bar, 20 μm.

Figure 1. — The brains of neonatal mice and preterm infants expressed A₁R receptors. (A) Immunostaining results showed the specificity of A₁R antibody but not A_2AR antibody. Using the tested A₁R antibody, A₁R immunostaining showed a punctate distribution of this receptor in the cortex of wild type adult mice, but not in that of A₁R deficient mice (25.33±2.4 and 8.67±0.88 puncta, respectively) (top). In contrast, several antibodies against A_2AR showed non-specific immunoreactivity in the nucleus and the cytoplasm in both wild type and A_2AR deficient mice (bottom). No punctate pattern was observed (6±1.53 puncta for WT and 4.33±0.3 puncta for A_2AR knockout). Representative images are shown here. The quantitative results are shown on the right. Blue arrows indicate puncta. Scale bar, 10 μm. (B) The punctate pattern of A₁R could be detected in the cortex of P3 pups. The A₁R puncta appeared more discrete at a lower magnification as the animals mature (10 day-old and 1 month-old pups). (C) A₁R distributed in a punctate pattern in medulla, cerebellum, thalamus and hippocampus. The puncta were more prominent in medulla and cerebellum. Scale bar, 20 μm. (D) Immunostaining showed a punctate pattern of A₁R in the cortex of preterm infants at the corrected age of 26–27 weeks. Representative images of the cortex from 6 preterm brains are shown. Scale bar, 10 μm. (E) A₁R colocalized with a synaptic protein, PSD-95, in frozen sections of neonatal mouse brain (upper panel) and paraffin sections of preterm infant brains (lower panel). Scale bar, 20 μm.