Journal: Purinergic Signalling

Article Title: Sensory-sympathetic coupling in superior cervical ganglia after myocardial ischemic injury facilitates sympathoexcitatory action via P2X 7 receptor

doi: 10.1007/s11302-013-9367-2

Figure Lengend Snippet: Effects of oxATP or knockdown P2X 7 on systolic blood pressure, diastolic blood pressure, heart rate

Article Snippet: The primary antibodies and dilutions used were as follows: rabbit polyclonal anti-phospho-p44/42 MAPK (ERK1/2, 1:1,000, Cell Signal CO.) and rabbit polyclonal anti-p44/42 MAPK (ERK1/2, 1:1000, Cell Signal CO.) P2X 7 receptor small interference RNA treatment The small interference RNA (siRNA) specific for rat P2X 7 was purchased from Invitrogen (Carlsbad, CA). siRNA oligonucleotides targeted specifically to rat P2X 7 were used in this experiment [ 35 ].

Techniques:

Journal: Purinergic Signalling

Article Title: Sensory-sympathetic coupling in superior cervical ganglia after myocardial ischemic injury facilitates sympathoexcitatory action via P2X 7 receptor

doi: 10.1007/s11302-013-9367-2

Figure Lengend Snippet: Effects of oxATP or knockdown P2X 7 on the serum concentration of CK-MB, CK, LDH, and cTn-I

Article Snippet: The primary antibodies and dilutions used were as follows: rabbit polyclonal anti-phospho-p44/42 MAPK (ERK1/2, 1:1,000, Cell Signal CO.) and rabbit polyclonal anti-p44/42 MAPK (ERK1/2, 1:1000, Cell Signal CO.) P2X 7 receptor small interference RNA treatment The small interference RNA (siRNA) specific for rat P2X 7 was purchased from Invitrogen (Carlsbad, CA). siRNA oligonucleotides targeted specifically to rat P2X 7 were used in this experiment [ 35 ].

Techniques: Concentration Assay

Journal: International Journal of Biological Sciences

Article Title: PRG-1 relieves pain and depressive-like behaviors in rats of bone cancer pain by regulation of dendritic spine in hippocampus

doi: 10.7150/ijbs.59032

Figure Lengend Snippet: Activation of hippocampal PRG-1 and P2X7 receptor in BCP rats. (A) DAPI overview of a rat showing areas of CA1 (highlighted by white square). (B) Images of PRG-1 (red) and P2X 7 receptor (green) in the hippocampus from sham and BCP rats by immunofluorescence on the POD 45 (scale bar = 50 μm). (C) Band of western blot and (D-E) quantitative analysis showed that the expression levels of PRG-1 and P2X 7 receptor were increased in the hippocampus of BCP rats compared with corresponding sham group (n = 6, one sample t test). Values represent mean ± SEM. *** P < 0.001. (F-G) Quantitative analysis of qRT-PCR showed that the transcriptions of PRG-1 and P2X 7 receptor mRNA were not increased in the hippocampus of BCP rats. β-actin was included as a control. The data are expressed as the mean ± SEM (n = 6). F: Mann-Whithnes-U test, G: unpaired Student t test, ** P <0.01, *** P <0.001 compared to the sham group. BCP, bone cancer pain.

Article Snippet: For immunofluorescence, the sections were blocked and permeabilized, and then incubated with goat anti-rat P2X 7 receptors antibody (1:400, abcam, USA) or rabbit anti-rat PRG-1 antibody (1:500, synaptic system).

Techniques: Activation Assay, Immunofluorescence, Western Blot, Expressing, Quantitative RT-PCR

Journal: International Journal of Biological Sciences

Article Title: PRG-1 relieves pain and depressive-like behaviors in rats of bone cancer pain by regulation of dendritic spine in hippocampus

doi: 10.7150/ijbs.59032

Figure Lengend Snippet: PRG-1 attenuated pain and depression-like behaviors in BCP rats. (A) Schematic diagram of hippocampal microinjection (x = ±2.0 mm, y = -3.72 mm, and z = -3.0 mm). (B) Hippocampus injection of virus vector LV-Plppr4 (PRG-1 OE) and LV-P2X7R-RNAi (P2X 7 receptor KD) alleviated BCP-induced TWL decreases while LV-Plppr4-RNAi (PRG-1 KD) lead to early and intensified pain (n = 12; one-way ANOVA, *: versus Sham group at the same time point; #: versus BCP group at the same time point). (C) Hippocampus injection of virus vector LV-Plppr4 (PRG-1 OE) and LV-P2X7R-RNAi (P2X 7 receptor KD) alleviated BCP-induced MWT decreases while LV-Plppr4-RNAi (PRG-1 KD) lead to early and intensified pain (n = 12; one-way ANOVA, *: versus Sham group at the same time point; #: versus BCP group at the same time point). (D) PRG-1 OE reversed the preference of sucrose (n = 12; one-way ANOVA, #: versus BCP group at the same time point). (E) Multiple FTY720 administration attenuated BCP-induced thermal withdrawal latency decreases from day 39 to 43 (n = 12; one-way ANOVA, *: versus Sham group at the same time point; #: versus BCP group at the same time point); dotted lines indicate pharmacological treatment). (F) Enlarged graph of D from POD 38 to 44. (G) Chronic FTY720 administration attenuated BCP-induced MWT decreases on day 40. (n = 6; Kruskal-Wallis test. #: versus BCP group at the same time point); dotted lines indicate pharmacological treatment). (H) FTY720 reversed the decline in the preference of sucrose consumption induced by BCP (n = 12 one-way ANOVA, *: versus Sham group at the same time point; #: versus BCP group at the same time point). The data are presented mean ± SEM. * P <0.05, ** P <0.01, *** P <0.001; BCP, bone cancer pain; TWL, thermal withdrawal latency; MWT, mechanical withdrawal threshold; OE, overexpression; KD, knock down.

Article Snippet: For immunofluorescence, the sections were blocked and permeabilized, and then incubated with goat anti-rat P2X 7 receptors antibody (1:400, abcam, USA) or rabbit anti-rat PRG-1 antibody (1:500, synaptic system).

Techniques: Injection, Plasmid Preparation, Over Expression

Journal: International Journal of Biological Sciences

Article Title: PRG-1 relieves pain and depressive-like behaviors in rats of bone cancer pain by regulation of dendritic spine in hippocampus

doi: 10.7150/ijbs.59032

Figure Lengend Snippet: PRG-1 attenuated neuron deactivation and synaptic depression in the hippocampus induced by BCP. (A) Nissl staining in CA1, DG districts of rats' hippocampus under FTY720 or A438079 treatment. (B) The graph shows the percent of nissl positive cell. n = 6; one-way ANOVA, *: versus Sham group. (C) Images of Golgi-Cox staining and (D-F) quantitative analysis showed that the spine density depression induced by BCP was reversed by FTY720. (D, n =20 Sham, 12 BCP, 14 BCP+FTY720 and 13 BCP+A438079, one-way ANOVA; E, n = 20 Sham,11 BCP, 12 BCP+FTY720 and 12 BCP+A438079, one-way ANOVA; F, n = 12 Sham, 10 BCP, 15 BCP+FTY720 and 15 BCP+A438079, Kruskal-Wallis test; n represents analyzed dendritic segments; *: versus Sham group; #: versus BCP group); stratum radiatum (sr, apical dendrites) and stratum oriens (so, basal dendrites) of the CA1 region, and the stratum moleculare of the dentate gyrus (moDG). (G) PRG-1 OE and P2X 7 R KD rescued nissl bodies and nucleoli in CA1 and DG regions. The data are expressed as the mean ± SEM; * P <0.05, ** P <0.01, *** P <0.001, BCP, bone cancer pain; OE, overexpression; KD, knock down.

Article Snippet: For immunofluorescence, the sections were blocked and permeabilized, and then incubated with goat anti-rat P2X 7 receptors antibody (1:400, abcam, USA) or rabbit anti-rat PRG-1 antibody (1:500, synaptic system).

Techniques: Staining, Over Expression

Journal: International Journal of Biological Sciences

Article Title: PRG-1 relieves pain and depressive-like behaviors in rats of bone cancer pain by regulation of dendritic spine in hippocampus

doi: 10.7150/ijbs.59032

Figure Lengend Snippet: PRG-1/PP2A interaction were increased by hippocampal injections of FTY720. (A) Images of PRG-1 (red) and P2X 7 receptor (green) in the hippocampus from BCP, BCP+FTY720 and BCP+A438079 rats by immunofluorescence on the POD 44 (scale bar = 50 μm). (B) Western blot showed that the expression of PRG-1 and P2X 7 receptor was increased in the hippocampus of BCP+FTY720 rats. (C) Co-immunoprecipitation (IP) using a PRG-1 antibody shows PRG-1 and PP2A association. (D-E) Quantitative analysis of RT-qPCR showed that the transcription of PRG-1 and P2X7 receptor mRNA was increased in the hippocampus of BCP+FTY720 rats. β-actin was included as a control. The data are expressed as the mean ± SEM; n = 6; D: Kruskal-Wallis test, E: one-way ANOVA; * P <0.05, ** P <0.005, *** P <0.001 compared to the BCP group. (F) Band of western blot showed the expression of PRG-1 and P2X 7 receptor induced by virus expression vectors. BCP, bone cancer pain; OE, overexpression; KD, knock down; POD, postoperative day.

Article Snippet: For immunofluorescence, the sections were blocked and permeabilized, and then incubated with goat anti-rat P2X 7 receptors antibody (1:400, abcam, USA) or rabbit anti-rat PRG-1 antibody (1:500, synaptic system).

Techniques: Immunofluorescence, Western Blot, Expressing, Immunoprecipitation, Quantitative RT-PCR, Over Expression