Journal: bioRxiv

Article Title: Adenosine deficiency facilitates CA1 synaptic hyperexcitability in the presymptomatic phase of a knock in mouse model of Alzheimer’s disease

doi: 10.1101/2024.04.24.590882

Figure Lengend Snippet: A) Microdialysis and quantification protocol. B) Mean adenosine levels C) and relationship between adenosine concentration and percentage of activity during dialysate collection scored by video monitoring the mice (WT: n=7, APPKI: n= 5.). D) Cartoon of the main metabolic reactions and enzymes of adenosine pathway. E) Representative western blots and F) mean normalized expression and single data points. (CD39: WT: n=8, APPKI: n= 7; CD73: WT: n=8, APPKI: n= 7; ENT1: WT: n=7, APPKI: n= 8; SAHH: WT: n=8, APPKI: n= 11; ADA: n=9, APPKI: n= 12; ADK: WT: n=8, APPKI: n= 10; A1R: WT: n=6, APPKI: n= 6). G) Left: Scheme of experiment to monitor SC-to-CA1 synaptic signaling (S1: stimulation; Rec: recording). Right: cartoon of adenosine signaling and inhibitor targets (cocktail: ARL 67156 trisodium salt: 50 µM; PSB 12379: 50 nM; PPADS tetrasodium salt: 50 µM; NBMPR: 300 nM). H) Left: Time trajectory of normalized fEPSP before (a) and after (b) the dosage of the drug cocktail. Right: Representative traces before (a) and after (b) the dosage of the drug cocktail. I) Mean FV-fEPSP slopes before (a) and after (b) drug cocktail and individual data points; WT: n=8/8, APPKI: n= 6/6.p: p-value; d : Cohen’s effect size. Mean±S.D. n=n. slices/n. mice. Statistic: generalized linear mixed effect models. Black: WT, Green: APPKI. Grey: WT+cocktail, Light Green: APPKI+cocktail.

Article Snippet: PPADS tetrasodium salt (50 μM; Tocris Bioscience) has been used as non-selective P2 antagonist to prevent the effect of ATP increase in response to the block of the ATP-to-adenosine conversion by PSB+ARL.

Techniques: Concentration Assay, Activity Assay, Western Blot, Expressing

Journal: Nature Communications

Article Title: A divergent astrocytic response to stress alters activity patterns via distinct mechanisms in male and female mice

doi: 10.1038/s41467-025-61643-y

Figure Lengend Snippet: A Schematic of orexin neuron tripartite synapse and overview of mechanism of action of pharmacological agents. B Percentage change in firing after bath application of modulators of the purinergic system. two-way ANOVA; Uncorrected Fisher’s LSD: 8-cyclopentyltheophylline ( CPT ) ( p = 0.0786, Naïve N = 2, n = 6 cells, ELS N = 2, n = 7 cells), ZM-241385 ( p = 0.973, Naïve N = 2, n = 6 cells, ELS N = 3, n = 5 cells), PSB12379 ( p < 0.001, Naïve N = 2, n = 6 cells, ELS N = 3, n = 6 cells), PPADS ( p = 0.0102, Naïve N = 2, n = 6 cells, ELS N = 3, n = 7 cells), iso-PPADS ( p < 0.001, Naïve N = 3, n = 5 cells, ELS N = 3, n = 7 cells). C Representative traces of orexin neurons from Naïve and ELS male mice before and after bath application of adenosine A1 receptor antagonist, CPT (200 nM). D Change in firing rate in Naïve mice with application of adenosine A1 receptor antagonist, CPT (Paired t test: baseline= 3.035, ±0.822, CPT (200 nM) = 1.56 ±0.564, ( N = 2, n = 6 cells) p = 0.0178. E Change in firing rate in ELS mice with application of CPT (Paired t test: baseline = 5.60 Hz ±1.47, CPT = 4.44 Hz ±1.6 9 ( N = 2, n = 7 cells) p = 0.068). F Representative traces of orexin neurons from Naïve and ELS male mice before and after bath application of adenosine A2 receptor antagonist, ZM241385 (50 nM). G Change in firing rate in Naïve mice with ZM241385 (Paired t test: baseline= 2.671 Hz ±0.461, ZM241385 = 2.422 Hz ±0.468 ( N = 2, n = 6 cells) p = 0.448). H Change in firing rate in ELS mice with application of adenosine A2 receptor antagonist, ZM241385 (Paired t test: baseline = 5.756 Hz ±0.938, ZM241385 = 5.421 Hz ±1.15 ( N = 3, n = 5 cells) p = 0.528). I Representative traces of orexin neurons from Naïve and ELS male mice before and after bath application of Ecto-5’-Nucleotidase (CD73) inhibitor, PSB12379 (10 μM). J Change in firing rate in Naïve mice with PSB12379 (Paired t test: baseline = 2.78 Hz ±0.826, PSB12379 = 5.29 Hz ±1.39 ( N = 2, n = 6 cells) p = 0.0086). K Change in firing rate in ELS mice with PSB12379 (Paired t test: baseline = 5.23 Hz ±0.720, PSB12379 = 6.105 Hz ±6.11 ( N = 3, n = 6 cells) p = 0.029). L Representative traces of orexin neurons from Naïve and ELS male mice before and after bath application of PPADS tetrasodium salt (50 μM). M Change in firing rate in Naïve mice with PPADS (Paired t test: baseline= 2.13 Hz ±0.440, PPADS = 1.88 Hz ±0.361 ( N = 2, n = 6 cells) p = 0.111). N Change in firing rate in ELS mice with PPADS (Paired t test: baseline= 6.52 Hz ±1.34, PPADS = 3.32 Hz ±0.737 ( N = 3, n = 7 cells) p = 0.025). O Representative trace of orexin neurons from ELS male mice before and after bath application of P2X purinoceptor antagonist, iso- PPADS tetrasodium salt (50 μM). P Change in firing rate in Naïve mice with iso- PPADS (Paired t test: baseline = 4.13 Hz ±0.368, iso- PPADS = 4.09 Hz ±0.377 ( N = 3, n = 5 cells) p = 0.237). Q Change in firing rate in ELS mice with iso- PPADS (Paired t test: baseline= 4.96 Hz ±0.759, iso- PPADS = 1.77 Hz ±0.448 ( N = 3, n = 7 cells) p = 0.006). R Zoom in representative trace after application of iso- PPADS. S Schematic highlighting shift in extracellular adenosine to ATP in ELS male mice. Extracellular ATP signalling through P2X receptors on the post-synaptic membrane to induce elevated spontaneous firing rates in orexin neurons. N = mice, n = cells. Bar charts = mean ± S.E.M. ns = not significant, * P < 0.05, ** P < 0.01.

Article Snippet: Purinergic 2 receptor antagonist, PPADS tetrasodium salt ( Tocris , 0625), and purinergic 2X receptor antagonist, iso -PPADS tetrasodium salt (Tocris, 0683).

Techniques: Membrane