Journal: Journal of pharmacological sciences

Article Title: P2Y 2 receptor mediates dying cell removal via inflammatory activated microglia.

doi: 10.1016/j.jphs.2023.06.004

Figure Lengend Snippet: Fig. 4. Effects of AR-C118925, a selective P2Y2 receptor antagonist, on the removal of dying cells by neighboring microglia. (A, B) Time-lapse observation of spontaneous (A) and LPS- induced microglial death (B) and removal of dying cells in the presence of AR-C118925. Microglia were treated with AR-C118925 (30 mM) and images captured every 10 min for 18 h. Images are representative of five independent experiments. Dying cells (white arrowhead); phagocytotic cells (black arrowhead). Time count, hr:min; scale bar, 50 mm, bottom. Movies 7 and 8 show representative live images. (C) Quantification of the removal of dying cells by control and AR-C118925-treated microglia. Fifty cells were randomly selected and manually divided into four groups based on their behavior. Group I, cells that underwent death; Group II, cells that did not make contact with any dying cells; Group III, cells that

Article Snippet: For livecell immunostaining, microglia were incubated with a FITCconjugated antibody against the extracellular domain of the P2Y2 receptor (1:50, ARP-102-F; Alomone) for 30 min on ice and then fixed with 4% paraformaldehyde for 15 min. Immunofluorescence staining of the P2Y2 receptor on the plasma membrane was observed with a fluorescence microscope (BIOREVO Bz-9000; Keyence).

Techniques: Control

Journal: Journal of pharmacological sciences

Article Title: P2Y 2 receptor mediates dying cell removal via inflammatory activated microglia.

doi: 10.1016/j.jphs.2023.06.004

Figure Lengend Snippet: Fig. 5. LPS stimulation caused a change in the intracellular localization of P2Y2 receptors in microglia. (A) Confocal micrographs of the P2Y2 receptor. Microglia were stimulated with LPS (10 ng/mL) for 3 h. After fixation and permeabilization, the cells were stained with an anti-P2Y2 receptor antibody, which recognizes the intracellular domain of the P2Y2 receptor, followed by Alexa Fluor 488-conjugated rabbit IgG as a secondary antibody, and fluorescence micrographs captured by using a confocal laser scanning microscope. White arrows indicate accumulation of the P2Y2 receptor along the edge of the plasma membrane. Experiments were repeated three times. Images are representative of three independent experiments. Scale bar, 50 mm. (B) Live cell images of the P2Y2 receptor on the plasma membrane of microglia. Microglia were stimulated with LPS for 3 h and stained with a FITC- conjugated antibody against the extracellular domain of the P2Y2 receptor for 30 min on ice. White arrowheads indicate the edge of the plasma membrane. Experiments were repeated three times. Images are representative of three independent experiments. Scale bar, 50 mm.

Article Snippet: For livecell immunostaining, microglia were incubated with a FITCconjugated antibody against the extracellular domain of the P2Y2 receptor (1:50, ARP-102-F; Alomone) for 30 min on ice and then fixed with 4% paraformaldehyde for 15 min. Immunofluorescence staining of the P2Y2 receptor on the plasma membrane was observed with a fluorescence microscope (BIOREVO Bz-9000; Keyence).

Techniques: Staining, Laser-Scanning Microscopy, Clinical Proteomics, Membrane

Journal: Journal of pharmacological sciences

Article Title: P2Y 2 receptor mediates dying cell removal via inflammatory activated microglia.

doi: 10.1016/j.jphs.2023.06.004

Figure Lengend Snippet: Fig. 6. Change in mRNA expression of the P2Y1, P2Y2, P2Y4, P2Y6, and P2Y12 receptors in microglia after LPS stimulation. Microglia were stimulated with LPS (10 ng/mL) for the indicated time. The mRNA expression level of each P2Y receptor was measured using real-time PCR and normalized to that of b-actin. Data are presented as the means ± SEM of independent experiments. n ¼ 10 for P2Y2; n ¼ 4 for P2Y1, P2Y4, P2Y6, and P2Y12.

Article Snippet: For livecell immunostaining, microglia were incubated with a FITCconjugated antibody against the extracellular domain of the P2Y2 receptor (1:50, ARP-102-F; Alomone) for 30 min on ice and then fixed with 4% paraformaldehyde for 15 min. Immunofluorescence staining of the P2Y2 receptor on the plasma membrane was observed with a fluorescence microscope (BIOREVO Bz-9000; Keyence).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Journal of pharmacological sciences

Article Title: P2Y 2 receptor mediates dying cell removal via inflammatory activated microglia.

doi: 10.1016/j.jphs.2023.06.004

Figure Lengend Snippet: Fig. 7. LPS induces Axl upregulation in a P2Y2 receptor-dependent manner in microglia. (A) LPS stimulation enhanced Axl mRNA expression (left) and suppressed that of Mer (right) in microglia. Microglia were stimulated with LPS (30 ng/mL) for the indicated time, and mRNA expression measured using real-time PCR. Values are expressed as the ratio of Mer or Axl to b-actin. Data are presented as the mean ± SEM of three independent experiments. (B) Effects of the selective P2Y2 receptor antagonist, AR-C118925, and the Pyk2 inhibitor, PF431396, on LPS-induced Axl mRNA upregulation in microglia. Microglia were treated with AR-C118925 (10, 30 mM) for 5 min and PF431396 (0.3, 1 mM) for 15 min and then stimulated with LPS (30 ng/mL) for 6 h. Data are presented as the means ± SEM (%) of LPS-induced Axl mRNA expression (vs. b-actin). n ¼ 5 for AR-C118925; n ¼ 3 for PF431396. C. Effects of AR-C118925 on LPS-induced Pyk2 upregulation in microglia. The mRNA expression levels were measured using real-time PCR and normalized to that of b-actin. Data are presented as the mean ± SEM of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 vs. LPS alone.

Article Snippet: For livecell immunostaining, microglia were incubated with a FITCconjugated antibody against the extracellular domain of the P2Y2 receptor (1:50, ARP-102-F; Alomone) for 30 min on ice and then fixed with 4% paraformaldehyde for 15 min. Immunofluorescence staining of the P2Y2 receptor on the plasma membrane was observed with a fluorescence microscope (BIOREVO Bz-9000; Keyence).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Pakistan Journal of Zoology

Article Title: Modulation of Pentylenetetrazol Induced Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats

doi: 10.17582/journal.pjz/20230402150443

Figure Lengend Snippet: Fig. 1. The P2Y3 antagonist NF45 has an effect on the mean kindling score of PTZ-kindling animals. (A) Seizure stage for each group. (B) Days of stimulation for each group at various phases. (C) Each group’s average kindling score.

Article Snippet: 2599 Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats 2599 The pro-inflammatory cytokines (TNF-α and 1L1β) have pro-epileptic properties and play a role in the induction and maintenance of seizures.

Techniques:

Journal: Pakistan Journal of Zoology

Article Title: Modulation of Pentylenetetrazol Induced Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats

doi: 10.17582/journal.pjz/20230402150443

Figure Lengend Snippet: Fig. 2. The P2Y3 antagonist NF45 has an effect on the mean kindling score of PTZ-kindling animals. (A) Seizure stage for each group. (B) Days of stimulation for each group at various phases. Group I, No treatment; Group II, infused with artificial cerebrospinal fluid; Group III, administered with NF 45; Group IV, administered with PTZ; Group V, administered with PTZ+30nm NF45; Group VI, administered with PTZ+60nm NF45; Group VII, administered with PTZ+90nM NF45; Group VIII, α-β- methylene-ATP+90nM NF45+PTZ.

Article Snippet: 2599 Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats 2599 The pro-inflammatory cytokines (TNF-α and 1L1β) have pro-epileptic properties and play a role in the induction and maintenance of seizures.

Techniques:

Journal: Pakistan Journal of Zoology

Article Title: Modulation of Pentylenetetrazol Induced Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats

doi: 10.17582/journal.pjz/20230402150443

Figure Lengend Snippet: Fig. 3. The P2Y3 antagonist NF45 has an effect on the anxiety behaviour of PTZ-kindling animals. (A) The amount of time spent in open and closed arms. (B) Frequency in the open and closed arm. For details of groups, see Figure 2.

Article Snippet: 2599 Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats 2599 The pro-inflammatory cytokines (TNF-α and 1L1β) have pro-epileptic properties and play a role in the induction and maintenance of seizures.

Techniques:

Journal: Pakistan Journal of Zoology

Article Title: Modulation of Pentylenetetrazol Induced Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats

doi: 10.17582/journal.pjz/20230402150443

Figure Lengend Snippet: Fig. 4. The effect of the P2Y3 antagonist NF45 on the ability of PTZ-kindling animals to discriminate. (A) The exploration time was identical in all groups in test 1 (T1), where both objects were similar or familiar (DO1 and DO2). (B) NF45 improved the separation of FO1 and NO significantly. For details of groups, see Figure 2.

Article Snippet: 2599 Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats 2599 The pro-inflammatory cytokines (TNF-α and 1L1β) have pro-epileptic properties and play a role in the induction and maintenance of seizures.

Techniques:

Journal: Pakistan Journal of Zoology

Article Title: Modulation of Pentylenetetrazol Induced Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats

doi: 10.17582/journal.pjz/20230402150443

Figure Lengend Snippet: Fig. 5. Effect of the P2Y3 antagonist NF45 on learning and memory of PTZ-kindling animals, as assessed on Morris water maze. In a dose-dependent manner, NF45 raised the TSTQ and number of crossings in the platform area of PTZ-kindling animals on day 5. For details of groups, see Figure 2.

Article Snippet: 2599 Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats 2599 The pro-inflammatory cytokines (TNF-α and 1L1β) have pro-epileptic properties and play a role in the induction and maintenance of seizures.

Techniques:

Journal: Pakistan Journal of Zoology

Article Title: Modulation of Pentylenetetrazol Induced Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats

doi: 10.17582/journal.pjz/20230402150443

Figure Lengend Snippet: Fig. 6. Effect of the P2Y3 antagonist NF45 on learning and memory of PTZ-kindling animals, as assessed on Morris water maze. (A) PTZ impaired learning of rats and NF45 mitigated this damage in PTZ-kindling animals. (B, C) In a dose-dependent manner, NF45 raised the TSTQ and number of crossings in the platform area of PTZ-kindling animals on day 5. For details of groups, see Figure 2.

Article Snippet: 2599 Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats 2599 The pro-inflammatory cytokines (TNF-α and 1L1β) have pro-epileptic properties and play a role in the induction and maintenance of seizures.

Techniques:

Journal: Pakistan Journal of Zoology

Article Title: Modulation of Pentylenetetrazol Induced Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats

doi: 10.17582/journal.pjz/20230402150443

Figure Lengend Snippet: Fig. 7. Effect of the P2Y3 antagonist NF110 on PTZ-kindling animals’ hippocampal IL-1β and TNF-α levels. (A) In PTZ-kindling animals, NF45 significantly reduced hippocampal IL-1β. (B) In PTZ-kindling rats, NF110 dramatically lowered hippocampus TNF-α levels. For details of groups, see Figure 2.

Article Snippet: 2599 Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats 2599 The pro-inflammatory cytokines (TNF-α and 1L1β) have pro-epileptic properties and play a role in the induction and maintenance of seizures.

Techniques:

Journal: Pakistan Journal of Zoology

Article Title: Modulation of Pentylenetetrazol Induced Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats

doi: 10.17582/journal.pjz/20230402150443

Figure Lengend Snippet: Fig. 8. Effect of the P2Y3 antagonist NF45 on hippocampal mitochondrial complex I, II, and IV levels of PTZ-kindling animals. NF45 significantly increased the hippocampal mitochondrial complex I (A), II (B), and IV (C) levels of PTZ-kindling animals.

Article Snippet: 2599 Neuronal Circuit Epilepsy by p2y Purinoreceptor 3 (p2y3) in Sprague Dawley Rats 2599 The pro-inflammatory cytokines (TNF-α and 1L1β) have pro-epileptic properties and play a role in the induction and maintenance of seizures.

Techniques: