Journal: International journal of molecular sciences

Article Title: Guanosine-Mediated Anxiolytic-Like Effect: Interplay with Adenosine A 1 and A 2A Receptors.

doi: 10.3390/ijms21239281

Figure Lengend Snippet: Figure 2. Functional competition between GUO and adenosine (ADO) or selective agonists for A1R and A2AR on GUO-mediated anxiolytic-like effect evaluated by EPM test during a 5 min session. Pretreatment with ADO (10 mg/kg or 30 mg/kg) resulted in a complete block of anxiolytic-like effect of GUO (30 mg/kg) with significant decrease in (A) number of open arm entries [F(5,62) =6.972, p < 0.0001], (B) time spent in open arms [F(5,62) = 6.566, p < 0.0001] and (C) the ratio of time in open arm and total time in arms [F(5,62) = 18.33, p < 0.0001], when compared to GUO treated group. Pretreatment with A1R agonist CPA (0.1 mg/kg) or with A2AR agonist CGS (0.4 mg/kg) was able to completely block the anxiolytic-like effect of GUO (30 mg/kg) with significant decrease in (D) number of open arm entries [F(5,60) = 5.450, p < 0.0005], (E) time spent in open arms [F(5,60) = 22.29, p < 0.0001] and (F) the ratio of time in open arm and total time in arms [F(5,60) = 15.54, p < 0.0001], when compared to GUO treated group. CGS alone produced a significant reduction in time spent in open arms and ratio of time in open arm and total time in arms as compared to control group (Ctrl): each bar represents the mean value ± SD. Tukey test: * p < 0.05, ** p < 0.01; *** p < 0.001. Ctrl raw mean values: (A) 6.23; (B) 76.23 s; (C) 0.33; (D) 7; (E) 97.88 s; (F) 0.43.

Article Snippet: The following drugs were used: GUO (G6264 Sigma-Aldrich St. Louis, MO, USA) and nonselective ARs ligand ADO (A3377 Sigma-Aldrich St. Louis, MO, USA) were dissolved in 0.9% physiological saline solution (pH 9.0); selective A1R agonists N6-Cyclopentyladenosine (CPA, C8031 Sigma-Aldrich) and 2-Chloro-N6-cyclopentyladenosine (CCPA, 1705 Tocris) or A2AR agonist 2-p-(2- Carboxyethyl)phenethylamino-5′-N-ethylcarboxamidoadenosine hydrochloride (CGS21680 hydrocloride, sc-211062 Santa Cruz Biotechnology) as well as selective A1R antagonist 8-Cyclopentyl-1,3dipropylxanthine (DPCPX, sc-200115 Santa-Cruz Biotechnology) or A2AR antagonist 4-(-2-[7-amino2-{2-furyl}triazolo [57] triazin-5-yl-amino]ethyl)phenol (ZM241385, Z0153, Sigma-Aldrich) were first dissolved in DMSO and thereafter in 0.9% physiological saline solution; the ARs nonselective antagonist caffeine (C0750, Sigma-Aldrich) was dissolved in 0.9% physiological saline solution.

Techniques: Functional Assay, Blocking Assay, Produced, Control

Journal: International journal of molecular sciences

Article Title: Guanosine-Mediated Anxiolytic-Like Effect: Interplay with Adenosine A 1 and A 2A Receptors.

doi: 10.3390/ijms21239281

Figure Lengend Snippet: Figure 3. Effects of A1R and A2AR antagonists on anxiolytic-like effect of GUO evaluated by EPM test during a 5 min session. (A–C) Functional competition between GUO and nonselective ARs antagonist caffeine (Caff). Caffeine alone (30 mg/kg) induced anxiolytic effects, as shown by significant increase in time spent in open arms and ratio of time in open arms and total time in arms as compared to control group (Ctrl). Pretreatment with caffeine (30 mg/kg) did not block anxiolytic-like effect of GUO and showed significant increase in time in open arms and ratio of time in open arms and total time in arms as compared to control group. (A–C) Functional competition between caffeine (30 mg/kg) and ADO (30 mg/kg) showed preservation of caffeine anxiolytic effect with significant increase in time spent in open arms and ratio of time in open arms and total time in arms as compared to control group. (D–F) Functional competition between GUO and selective AR antagonists. Selective A1R antagonist DPCPX (1 mg/kg) was not able to block GUO anxiolytic-like effect, as shown by significant increase in time in open arms and ratio of time in open arms and total time in arms as compared to control group. DPCPX alone did not show significant changes of all EPM parameters as compared to control group. Pretreatment with selective A2AR antagonist ZM241385 (0.4 mg/kg) was not able to block GUO (30 mg/kg) anxiolytic-like effect, as shown by significant increase in time in open arms and ratio of time in open arms and total time in arms as compared to control group, and ZM241385 alone did not show

Article Snippet: The following drugs were used: GUO (G6264 Sigma-Aldrich St. Louis, MO, USA) and nonselective ARs ligand ADO (A3377 Sigma-Aldrich St. Louis, MO, USA) were dissolved in 0.9% physiological saline solution (pH 9.0); selective A1R agonists N6-Cyclopentyladenosine (CPA, C8031 Sigma-Aldrich) and 2-Chloro-N6-cyclopentyladenosine (CCPA, 1705 Tocris) or A2AR agonist 2-p-(2- Carboxyethyl)phenethylamino-5′-N-ethylcarboxamidoadenosine hydrochloride (CGS21680 hydrocloride, sc-211062 Santa Cruz Biotechnology) as well as selective A1R antagonist 8-Cyclopentyl-1,3dipropylxanthine (DPCPX, sc-200115 Santa-Cruz Biotechnology) or A2AR antagonist 4-(-2-[7-amino2-{2-furyl}triazolo [57] triazin-5-yl-amino]ethyl)phenol (ZM241385, Z0153, Sigma-Aldrich) were first dissolved in DMSO and thereafter in 0.9% physiological saline solution; the ARs nonselective antagonist caffeine (C0750, Sigma-Aldrich) was dissolved in 0.9% physiological saline solution.

Techniques: Functional Assay, Control, Blocking Assay, Preserving

Journal: International journal of molecular sciences

Article Title: Guanosine-Mediated Anxiolytic-Like Effect: Interplay with Adenosine A 1 and A 2A Receptors.

doi: 10.3390/ijms21239281

Figure Lengend Snippet: Figure 5. [3H]GUO binding to hippocampal membranes. (A) The saturation isotherm studies showed that the binding became saturable at [3H]GUO concentrations ranging between 100 and 300 nM. The pooled data resolved for the presence of a single high affinity binding site with an apparent KD = 80 ± 34 nM; Bmax= 2339 ± 339 fmol/mg /protein. (B) Displacement of [3H]GUO binding by GUO and nonselective ARs agonist ADO in rat hippocampal membranes. Competition binding between GUO and ADO showed for ADO almost the same potency order of GUO to displace [3H]GUO (pIC50 6.069 ± 0.2074 and pIC50 −6.251 ± 0.1649, respectively), although ADO was able to displace only 70% of [3H]GUO binding. (C) [3H]GUO displacement (70 nM) by 500 µM of GUO, ADO, caffeine and selective agonists CPA or CGS21680 in rat hippocampal membranes.ADO was almost as effective as GUO in displacing [3H]GUO binding. Selective A1R agonist CPA or selective A2AR agonist CGS21680 displaced respectively 57% and 11% of [3H]GUO binding. Nonselective ARs antagonist caffeine displaced only 24% of [3H]GUO binding.

Article Snippet: The following drugs were used: GUO (G6264 Sigma-Aldrich St. Louis, MO, USA) and nonselective ARs ligand ADO (A3377 Sigma-Aldrich St. Louis, MO, USA) were dissolved in 0.9% physiological saline solution (pH 9.0); selective A1R agonists N6-Cyclopentyladenosine (CPA, C8031 Sigma-Aldrich) and 2-Chloro-N6-cyclopentyladenosine (CCPA, 1705 Tocris) or A2AR agonist 2-p-(2- Carboxyethyl)phenethylamino-5′-N-ethylcarboxamidoadenosine hydrochloride (CGS21680 hydrocloride, sc-211062 Santa Cruz Biotechnology) as well as selective A1R antagonist 8-Cyclopentyl-1,3dipropylxanthine (DPCPX, sc-200115 Santa-Cruz Biotechnology) or A2AR antagonist 4-(-2-[7-amino2-{2-furyl}triazolo [57] triazin-5-yl-amino]ethyl)phenol (ZM241385, Z0153, Sigma-Aldrich) were first dissolved in DMSO and thereafter in 0.9% physiological saline solution; the ARs nonselective antagonist caffeine (C0750, Sigma-Aldrich) was dissolved in 0.9% physiological saline solution.

Techniques: Binding Assay