p2x3r  (Alomone Labs)


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    Structured Review

    Alomone Labs p2x3r
    Expression of purinergic receptors and pannexin 1 channels is altered in T1D Akita mice bones. (A) Relative protein expression levels of P2R subtypes (P2Y 1 R, P2Y 2 R, P2Y 4 R, <t>P2X3R,</t> P2X4R and P2X7R) and Panx1 in 8-week-old Akita and age-matched wildtype bone tissue (Data are presented as the means ± SEM *P
    P2x3r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p2x3r/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
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    p2x3r - by Bioz Stars, 2022-07
    93/100 stars

    Images

    1) Product Images from "P2X7R-Panx1 Complex Impairs Bone Mechanosignaling under High Glucose Levels Associated with Type-1 Diabetes"

    Article Title: P2X7R-Panx1 Complex Impairs Bone Mechanosignaling under High Glucose Levels Associated with Type-1 Diabetes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0155107

    Expression of purinergic receptors and pannexin 1 channels is altered in T1D Akita mice bones. (A) Relative protein expression levels of P2R subtypes (P2Y 1 R, P2Y 2 R, P2Y 4 R, P2X3R, P2X4R and P2X7R) and Panx1 in 8-week-old Akita and age-matched wildtype bone tissue (Data are presented as the means ± SEM *P
    Figure Legend Snippet: Expression of purinergic receptors and pannexin 1 channels is altered in T1D Akita mice bones. (A) Relative protein expression levels of P2R subtypes (P2Y 1 R, P2Y 2 R, P2Y 4 R, P2X3R, P2X4R and P2X7R) and Panx1 in 8-week-old Akita and age-matched wildtype bone tissue (Data are presented as the means ± SEM *P

    Techniques Used: Expressing, Mouse Assay

    2) Product Images from "Species generalization and differences in Hedgehog pathway regulation of fungiform and circumvallate papilla taste function and somatosensation demonstrated with sonidegib"

    Article Title: Species generalization and differences in Hedgehog pathway regulation of fungiform and circumvallate papilla taste function and somatosensation demonstrated with sonidegib

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-34399-3

    Sonidegib treatment reduces taste buds (TB), SHH ligand and proliferation in rat fungiform papilla (FP) while innervation is retained. ( a ) Immunofluorescent antibody detection of SHH ligand (red) and K19 (green) for TB cells; K18 (red) for TB cells and Ki67 (green) for cell proliferation, after Vehicle, 16d or 28d Sonidegib treatments. SHH is reduced in association with TB, K19+ cell loss. Asterisks (*) indicate nonspecific SHH immunoproduct in cornified surface cells in 16d Sonidegib image. The Vehicle, K18/Ki67 image shows 3 regions positive for Ki67+ cells (Apical, Basal and Perigemmal). Proliferating cells are lost in Apical FP region after 16–28d Sonidegib. ( b ) Number of Ki67+ cells in Apical and Basal regions of FP in Vehicle- and Sonidegib-treated mice. Numbers of tongues analyzed are in parentheses. For each tongue 8–10 FP were analyzed. Sonidegib treatment reduces apical epithelial cell proliferation in FP compared to Vehicle. Statistical analysis was one-way ANOVA with Tukey HSD posthoc comparisons (*p ≤ 0.05, compared to Vehicle, APICAL). ( c ) Immunofluorescent antibody detection of K19 or K18 (red) for TB cells and NF (green) for lingual and CT innervation or P2X3 (green) for CT nerve fibers. Innervation was retained after Sonidegib exposure. Asterisks (*) indicate nonspecific P2X3 immunoproduct in surface layer in Vehicle image. ( d ) Enlarged images from 28d Sonidegib papillae. Arrows point to NF+ or P2X3+ fibers in the FP epithelium. Throughout, white dotted lines indicate the basal lamina. Yellow dotted lines indicate surface of epithelium. ( a , c ) Scale bar: 50 μm, applies to all images. ( d ) Scale bar: 25 μm, applies to both images.
    Figure Legend Snippet: Sonidegib treatment reduces taste buds (TB), SHH ligand and proliferation in rat fungiform papilla (FP) while innervation is retained. ( a ) Immunofluorescent antibody detection of SHH ligand (red) and K19 (green) for TB cells; K18 (red) for TB cells and Ki67 (green) for cell proliferation, after Vehicle, 16d or 28d Sonidegib treatments. SHH is reduced in association with TB, K19+ cell loss. Asterisks (*) indicate nonspecific SHH immunoproduct in cornified surface cells in 16d Sonidegib image. The Vehicle, K18/Ki67 image shows 3 regions positive for Ki67+ cells (Apical, Basal and Perigemmal). Proliferating cells are lost in Apical FP region after 16–28d Sonidegib. ( b ) Number of Ki67+ cells in Apical and Basal regions of FP in Vehicle- and Sonidegib-treated mice. Numbers of tongues analyzed are in parentheses. For each tongue 8–10 FP were analyzed. Sonidegib treatment reduces apical epithelial cell proliferation in FP compared to Vehicle. Statistical analysis was one-way ANOVA with Tukey HSD posthoc comparisons (*p ≤ 0.05, compared to Vehicle, APICAL). ( c ) Immunofluorescent antibody detection of K19 or K18 (red) for TB cells and NF (green) for lingual and CT innervation or P2X3 (green) for CT nerve fibers. Innervation was retained after Sonidegib exposure. Asterisks (*) indicate nonspecific P2X3 immunoproduct in surface layer in Vehicle image. ( d ) Enlarged images from 28d Sonidegib papillae. Arrows point to NF+ or P2X3+ fibers in the FP epithelium. Throughout, white dotted lines indicate the basal lamina. Yellow dotted lines indicate surface of epithelium. ( a , c ) Scale bar: 50 μm, applies to all images. ( d ) Scale bar: 25 μm, applies to both images.

    Techniques Used: Mouse Assay

    Long term Sonidegib treatment in mouse reduces taste buds (TB) and SHH ligand in circumvallate papilla (CV) whereas proliferation and innervation are retained. Immunofluorescent antibody detection of SHH ligand (red) and K8 (green) for TB cells; K8 (red) for TB cells and Ki67 (green) for cell proliferation; and K8 (red) with NF (green) for GL innervation or P2X3 (green) for GL taste fibers, after Vehicle or 48d Sonidegib treatment. For SHH/K8, large dotted lines indicate the basal lamina. Small dotted lines outline the surface epithelium. Inset (K8/P2X3) shows an image of nerves extending into CV epithelial basal lamina (arrow). Scale bar: 50 μm, applies to all images. Inset at 2×.
    Figure Legend Snippet: Long term Sonidegib treatment in mouse reduces taste buds (TB) and SHH ligand in circumvallate papilla (CV) whereas proliferation and innervation are retained. Immunofluorescent antibody detection of SHH ligand (red) and K8 (green) for TB cells; K8 (red) for TB cells and Ki67 (green) for cell proliferation; and K8 (red) with NF (green) for GL innervation or P2X3 (green) for GL taste fibers, after Vehicle or 48d Sonidegib treatment. For SHH/K8, large dotted lines indicate the basal lamina. Small dotted lines outline the surface epithelium. Inset (K8/P2X3) shows an image of nerves extending into CV epithelial basal lamina (arrow). Scale bar: 50 μm, applies to all images. Inset at 2×.

    Techniques Used:

    Sonidegib treatment in rat reduces taste buds (TB) and SHH ligand in circumvallate papilla (CV) whereas cell proliferation and GL innervation are retained. Immunofluorescent antibody detection of SHH ligand (red) and K19 (green) for TB cells; K18 (red) for TB cells and Ki67 (green) for cell proliferation; K19 (red) for TB cells and NF (green) for innervation; and, K18 (red) for TB cells and P2X3 (green) for taste nerve fibers, after Vehicle or 36d Sonidegib treatment. White dotted lines outline the epithelium. Asterisk in SHH/K19, 36d Sonidegib indicates nonspecific K19 immunostaining. Arrow points to the P2X3+ nerves extending into CV epithelium after Sonidegib treatment. SHH is reduced in association with TB cells. Cell proliferation is maintained and nerves fibers are retained after Sonidegib treatment. Scale bar: 50μm, applies to all images, except K19/NF.
    Figure Legend Snippet: Sonidegib treatment in rat reduces taste buds (TB) and SHH ligand in circumvallate papilla (CV) whereas cell proliferation and GL innervation are retained. Immunofluorescent antibody detection of SHH ligand (red) and K19 (green) for TB cells; K18 (red) for TB cells and Ki67 (green) for cell proliferation; K19 (red) for TB cells and NF (green) for innervation; and, K18 (red) for TB cells and P2X3 (green) for taste nerve fibers, after Vehicle or 36d Sonidegib treatment. White dotted lines outline the epithelium. Asterisk in SHH/K19, 36d Sonidegib indicates nonspecific K19 immunostaining. Arrow points to the P2X3+ nerves extending into CV epithelium after Sonidegib treatment. SHH is reduced in association with TB cells. Cell proliferation is maintained and nerves fibers are retained after Sonidegib treatment. Scale bar: 50μm, applies to all images, except K19/NF.

    Techniques Used: Immunostaining

    3) Product Images from "Species generalization and differences in Hedgehog pathway regulation of fungiform and circumvallate papilla taste function and somatosensation demonstrated with sonidegib"

    Article Title: Species generalization and differences in Hedgehog pathway regulation of fungiform and circumvallate papilla taste function and somatosensation demonstrated with sonidegib

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-34399-3

    Sonidegib treatment reduces taste buds (TB), SHH ligand and proliferation in rat fungiform papilla (FP) while innervation is retained. ( a ) Immunofluorescent antibody detection of SHH ligand (red) and K19 (green) for TB cells; K18 (red) for TB cells and Ki67 (green) for cell proliferation, after Vehicle, 16d or 28d Sonidegib treatments. SHH is reduced in association with TB, K19+ cell loss. Asterisks (*) indicate nonspecific SHH immunoproduct in cornified surface cells in 16d Sonidegib image. The Vehicle, K18/Ki67 image shows 3 regions positive for Ki67+ cells (Apical, Basal and Perigemmal). Proliferating cells are lost in Apical FP region after 16–28d Sonidegib. ( b ) Number of Ki67+ cells in Apical and Basal regions of FP in Vehicle- and Sonidegib-treated mice. Numbers of tongues analyzed are in parentheses. For each tongue 8–10 FP were analyzed. Sonidegib treatment reduces apical epithelial cell proliferation in FP compared to Vehicle. Statistical analysis was one-way ANOVA with Tukey HSD posthoc comparisons (*p ≤ 0.05, compared to Vehicle, APICAL). ( c ) Immunofluorescent antibody detection of K19 or K18 (red) for TB cells and NF (green) for lingual and CT innervation or P2X3 (green) for CT nerve fibers. Innervation was retained after Sonidegib exposure. Asterisks (*) indicate nonspecific P2X3 immunoproduct in surface layer in Vehicle image. ( d ) Enlarged images from 28d Sonidegib papillae. Arrows point to NF+ or P2X3+ fibers in the FP epithelium. Throughout, white dotted lines indicate the basal lamina. Yellow dotted lines indicate surface of epithelium. ( a , c ) Scale bar: 50 μm, applies to all images. ( d ) Scale bar: 25 μm, applies to both images.
    Figure Legend Snippet: Sonidegib treatment reduces taste buds (TB), SHH ligand and proliferation in rat fungiform papilla (FP) while innervation is retained. ( a ) Immunofluorescent antibody detection of SHH ligand (red) and K19 (green) for TB cells; K18 (red) for TB cells and Ki67 (green) for cell proliferation, after Vehicle, 16d or 28d Sonidegib treatments. SHH is reduced in association with TB, K19+ cell loss. Asterisks (*) indicate nonspecific SHH immunoproduct in cornified surface cells in 16d Sonidegib image. The Vehicle, K18/Ki67 image shows 3 regions positive for Ki67+ cells (Apical, Basal and Perigemmal). Proliferating cells are lost in Apical FP region after 16–28d Sonidegib. ( b ) Number of Ki67+ cells in Apical and Basal regions of FP in Vehicle- and Sonidegib-treated mice. Numbers of tongues analyzed are in parentheses. For each tongue 8–10 FP were analyzed. Sonidegib treatment reduces apical epithelial cell proliferation in FP compared to Vehicle. Statistical analysis was one-way ANOVA with Tukey HSD posthoc comparisons (*p ≤ 0.05, compared to Vehicle, APICAL). ( c ) Immunofluorescent antibody detection of K19 or K18 (red) for TB cells and NF (green) for lingual and CT innervation or P2X3 (green) for CT nerve fibers. Innervation was retained after Sonidegib exposure. Asterisks (*) indicate nonspecific P2X3 immunoproduct in surface layer in Vehicle image. ( d ) Enlarged images from 28d Sonidegib papillae. Arrows point to NF+ or P2X3+ fibers in the FP epithelium. Throughout, white dotted lines indicate the basal lamina. Yellow dotted lines indicate surface of epithelium. ( a , c ) Scale bar: 50 μm, applies to all images. ( d ) Scale bar: 25 μm, applies to both images.

    Techniques Used: Mouse Assay

    Long term Sonidegib treatment in mouse reduces taste buds (TB) and SHH ligand in circumvallate papilla (CV) whereas proliferation and innervation are retained. Immunofluorescent antibody detection of SHH ligand (red) and K8 (green) for TB cells; K8 (red) for TB cells and Ki67 (green) for cell proliferation; and K8 (red) with NF (green) for GL innervation or P2X3 (green) for GL taste fibers, after Vehicle or 48d Sonidegib treatment. For SHH/K8, large dotted lines indicate the basal lamina. Small dotted lines outline the surface epithelium. Inset (K8/P2X3) shows an image of nerves extending into CV epithelial basal lamina (arrow). Scale bar: 50 μm, applies to all images. Inset at 2×.
    Figure Legend Snippet: Long term Sonidegib treatment in mouse reduces taste buds (TB) and SHH ligand in circumvallate papilla (CV) whereas proliferation and innervation are retained. Immunofluorescent antibody detection of SHH ligand (red) and K8 (green) for TB cells; K8 (red) for TB cells and Ki67 (green) for cell proliferation; and K8 (red) with NF (green) for GL innervation or P2X3 (green) for GL taste fibers, after Vehicle or 48d Sonidegib treatment. For SHH/K8, large dotted lines indicate the basal lamina. Small dotted lines outline the surface epithelium. Inset (K8/P2X3) shows an image of nerves extending into CV epithelial basal lamina (arrow). Scale bar: 50 μm, applies to all images. Inset at 2×.

    Techniques Used:

    Sonidegib treatment in rat reduces taste buds (TB) and SHH ligand in circumvallate papilla (CV) whereas cell proliferation and GL innervation are retained. Immunofluorescent antibody detection of SHH ligand (red) and K19 (green) for TB cells; K18 (red) for TB cells and Ki67 (green) for cell proliferation; K19 (red) for TB cells and NF (green) for innervation; and, K18 (red) for TB cells and P2X3 (green) for taste nerve fibers, after Vehicle or 36d Sonidegib treatment. White dotted lines outline the epithelium. Asterisk in SHH/K19, 36d Sonidegib indicates nonspecific K19 immunostaining. Arrow points to the P2X3+ nerves extending into CV epithelium after Sonidegib treatment. SHH is reduced in association with TB cells. Cell proliferation is maintained and nerves fibers are retained after Sonidegib treatment. Scale bar: 50μm, applies to all images, except K19/NF.
    Figure Legend Snippet: Sonidegib treatment in rat reduces taste buds (TB) and SHH ligand in circumvallate papilla (CV) whereas cell proliferation and GL innervation are retained. Immunofluorescent antibody detection of SHH ligand (red) and K19 (green) for TB cells; K18 (red) for TB cells and Ki67 (green) for cell proliferation; K19 (red) for TB cells and NF (green) for innervation; and, K18 (red) for TB cells and P2X3 (green) for taste nerve fibers, after Vehicle or 36d Sonidegib treatment. White dotted lines outline the epithelium. Asterisk in SHH/K19, 36d Sonidegib indicates nonspecific K19 immunostaining. Arrow points to the P2X3+ nerves extending into CV epithelium after Sonidegib treatment. SHH is reduced in association with TB cells. Cell proliferation is maintained and nerves fibers are retained after Sonidegib treatment. Scale bar: 50μm, applies to all images, except K19/NF.

    Techniques Used: Immunostaining

    4) Product Images from "Altered expression of P2X3 in vagal and spinal afferents following esophagitis in rats"

    Article Title: Altered expression of P2X3 in vagal and spinal afferents following esophagitis in rats

    Journal: Histochemistry and cell biology

    doi: 10.1007/s00418-009-0639-4

    a Distribution pattern of P2X 3 immunoreactivity in peptidergic and non peptidergic small diameter c-fibers in DRG (T3) and NG. DRG sections from sham-operated rat with P2X3-SP immunostaining ( left column, upper panel ) and P2X3-IB4 staining ( left column,
    Figure Legend Snippet: a Distribution pattern of P2X 3 immunoreactivity in peptidergic and non peptidergic small diameter c-fibers in DRG (T3) and NG. DRG sections from sham-operated rat with P2X3-SP immunostaining ( left column, upper panel ) and P2X3-IB4 staining ( left column,

    Techniques Used: Immunostaining, Staining

    Photomicrography of P2X3 immunoreactivity in the NG and thoracic (T8) DRG retrogradely labeled with Fast Blue (FB). FB was injected in the subdiaphragmatic segment of the esophagus and FB positive cell bodies in the NG ( left column ) and DRG ( right column
    Figure Legend Snippet: Photomicrography of P2X3 immunoreactivity in the NG and thoracic (T8) DRG retrogradely labeled with Fast Blue (FB). FB was injected in the subdiaphragmatic segment of the esophagus and FB positive cell bodies in the NG ( left column ) and DRG ( right column

    Techniques Used: Labeling, Injection

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    Alomone Labs rabbit anti p2x3r
    Go6976 reduces α,β-meATP–elicited flinch nocifensive responses in complete Freund adjuvant–treated rats, and PKCβ1 is not expressed in <t>P2X3R-containing</t> dorsal root ganglia (DRG) neurons. (A) In saline control rats,
    Rabbit Anti P2x3r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti p2x3r/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti p2x3r - by Bioz Stars, 2022-07
    93/100 stars
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    Go6976 reduces α,β-meATP–elicited flinch nocifensive responses in complete Freund adjuvant–treated rats, and PKCβ1 is not expressed in P2X3R-containing dorsal root ganglia (DRG) neurons. (A) In saline control rats,

    Journal: Pain

    Article Title: Epac–protein kinase C alpha signaling in purinergic P2X3R-mediated hyperalgesia after inflammation

    doi: 10.1097/j.pain.0000000000000547

    Figure Lengend Snippet: Go6976 reduces α,β-meATP–elicited flinch nocifensive responses in complete Freund adjuvant–treated rats, and PKCβ1 is not expressed in P2X3R-containing dorsal root ganglia (DRG) neurons. (A) In saline control rats,

    Article Snippet: Antibodies used were mouse anti-Epac1, anti-Epac2 (1:1000; Cell Signaling, Danvers, MA), mouse anti-PKCε (1:1000; Santa Cruz), rabbit anti-pPKCε (1:1000; Santa Cruz), and rabbit anti-P2X3R (1:2000; Alomone Labs). β-Actin, probed with anti-β-actin (1:10,000; Chemicon, Temeculla, CA), or the neuronal marker, β-tubulin, probed with anti β-tubulin antibody (1:2000; Santa Cruz), was used as loading control.

    Techniques:

    Phosphorylated PKC (pPKC) isoform and P2X3R expression in dorsal root ganglia slices prepared from complete Freund adjuvant–treated rats. Upper: pPKCα and P2X3Rs were expressed only in small or medium cells. pPKCα labels were found

    Journal: Pain

    Article Title: Epac–protein kinase C alpha signaling in purinergic P2X3R-mediated hyperalgesia after inflammation

    doi: 10.1097/j.pain.0000000000000547

    Figure Lengend Snippet: Phosphorylated PKC (pPKC) isoform and P2X3R expression in dorsal root ganglia slices prepared from complete Freund adjuvant–treated rats. Upper: pPKCα and P2X3Rs were expressed only in small or medium cells. pPKCα labels were found

    Article Snippet: Antibodies used were mouse anti-Epac1, anti-Epac2 (1:1000; Cell Signaling, Danvers, MA), mouse anti-PKCε (1:1000; Santa Cruz), rabbit anti-pPKCε (1:1000; Santa Cruz), and rabbit anti-P2X3R (1:2000; Alomone Labs). β-Actin, probed with anti-β-actin (1:10,000; Chemicon, Temeculla, CA), or the neuronal marker, β-tubulin, probed with anti β-tubulin antibody (1:2000; Santa Cruz), was used as loading control.

    Techniques: Expressing

    ATP, P2X2, and P2X3 in human HNSCC microenvironment and pain. a . Representative HPLC chromatograms showing ATP peaks from tumor and matched normal tissue harvested from the same patient. b . Tumor tissues (Ipsi) had higher levels of ATP compared to matched normal sites (Contra) (n = 10, Student’s t -test). c . Pain scores of functional sharpness and intensity were significantly higher than spontaneous sharpness and intensity, respectively (n = 13, Student’s t -test). d . Mean scores of functional pain (Q2, 4, 6, 7, 8) were significantly higher than spontaneous pain (Q1, 3, 5) (Student’s t -test). e . ATP concentration in extracted cancer tissue correlated positively with mean pain scores (linear regression). f . Representative H E and immunofluorescence staining (P2X2, P2X3, merged) of a human tongue SCC. Sections were taken from adjacent sections of the SCC. Scale bar: 100 μm.

    Journal: Acta Neuropathologica Communications

    Article Title: Adenosine triphosphate drives head and neck cancer pain through P2X2/3 heterotrimers

    doi: 10.1186/2051-5960-2-62

    Figure Lengend Snippet: ATP, P2X2, and P2X3 in human HNSCC microenvironment and pain. a . Representative HPLC chromatograms showing ATP peaks from tumor and matched normal tissue harvested from the same patient. b . Tumor tissues (Ipsi) had higher levels of ATP compared to matched normal sites (Contra) (n = 10, Student’s t -test). c . Pain scores of functional sharpness and intensity were significantly higher than spontaneous sharpness and intensity, respectively (n = 13, Student’s t -test). d . Mean scores of functional pain (Q2, 4, 6, 7, 8) were significantly higher than spontaneous pain (Q1, 3, 5) (Student’s t -test). e . ATP concentration in extracted cancer tissue correlated positively with mean pain scores (linear regression). f . Representative H E and immunofluorescence staining (P2X2, P2X3, merged) of a human tongue SCC. Sections were taken from adjacent sections of the SCC. Scale bar: 100 μm.

    Article Snippet: For immunofluorescence labeling, neurons or tissue sections were then incubated for 24 h at 4°C in rabbit anti-P2X3 (1:500, Alomone Labs) and goat anti-P2X2 (1:500, Santa Cruz Biotechnology).

    Techniques: High Performance Liquid Chromatography, Functional Assay, Concentration Assay, Immunofluorescence, Staining

    HNSCC induces neuronal P2X2/3 plasticity that is reversed by anti-NGF. a . Sustained ATP current (top panel) is enhanced and prolonged following SCC co-culture; anti-NGF added into the co-culture reduced the sustained ATP current. Transient ATP current (lower panel) is not affected by either co-culture or anti-NGF. b . Sustained ATP current density is increased by co-culture (One-way ANOVA), and is reversed by anti-NGF. c . Dot plot of sustained ATP current in different diameter neurons. Co-culture increased current in medium-sized TG neurons; this increased current was reversed by anti-NGF. d . Representative immunofluorescence images of P2X2 and P2X3 expression in TG neurons. e . HNSCC co-culture increased the percentage of neurons expressing P2X2 and P2X3 subunits. Anti-NGF treatment significantly reduced percentage of neurons expressing P2X3 but not P2X2 subunits. The significant increase in the percentage of neurons that express both subunits following co-culture was reversed by anti-NGF application (One-way ANOVA). f . P2X2 immunofluorescence intensity was not changed after co-culture or anti-NGF treatment. P2X3 immunofluorescence intensity was significantly increased following co-culture, and was reduced by anti-NGF (one-way ANOVA). g . In mice with tongue HNSCC, mRNA expression for P2X2 was increased, while P2X3 expression was unchanged in TG neurons (Student’s t -test).

    Journal: Acta Neuropathologica Communications

    Article Title: Adenosine triphosphate drives head and neck cancer pain through P2X2/3 heterotrimers

    doi: 10.1186/2051-5960-2-62

    Figure Lengend Snippet: HNSCC induces neuronal P2X2/3 plasticity that is reversed by anti-NGF. a . Sustained ATP current (top panel) is enhanced and prolonged following SCC co-culture; anti-NGF added into the co-culture reduced the sustained ATP current. Transient ATP current (lower panel) is not affected by either co-culture or anti-NGF. b . Sustained ATP current density is increased by co-culture (One-way ANOVA), and is reversed by anti-NGF. c . Dot plot of sustained ATP current in different diameter neurons. Co-culture increased current in medium-sized TG neurons; this increased current was reversed by anti-NGF. d . Representative immunofluorescence images of P2X2 and P2X3 expression in TG neurons. e . HNSCC co-culture increased the percentage of neurons expressing P2X2 and P2X3 subunits. Anti-NGF treatment significantly reduced percentage of neurons expressing P2X3 but not P2X2 subunits. The significant increase in the percentage of neurons that express both subunits following co-culture was reversed by anti-NGF application (One-way ANOVA). f . P2X2 immunofluorescence intensity was not changed after co-culture or anti-NGF treatment. P2X3 immunofluorescence intensity was significantly increased following co-culture, and was reduced by anti-NGF (one-way ANOVA). g . In mice with tongue HNSCC, mRNA expression for P2X2 was increased, while P2X3 expression was unchanged in TG neurons (Student’s t -test).

    Article Snippet: For immunofluorescence labeling, neurons or tissue sections were then incubated for 24 h at 4°C in rabbit anti-P2X3 (1:500, Alomone Labs) and goat anti-P2X2 (1:500, Santa Cruz Biotechnology).

    Techniques: Co-Culture Assay, Immunofluorescence, Expressing, Mouse Assay

    Blocking ATP production in mitochondria blocks release of ATP Oligomycin blocks ATP synthase in mitochondria allowing for the build-up of ADP within the matrix and intermembrane spaces. (A) ATP biosensor cells were transfected with mRNAs encoding P2X2/P2X3 ionotropic purinergic receptors, which are responsive to ATP but not ADP. (B) Before application of oligomycin, taste cells show a robust and repeatable release of ATP (blue trace). Five minutes after application of oligomycin, depolarization-evoked ATP release was eliminated (red trace). The bar graph shows a quantitative comparison of the responses (5 minutes with oligomycin, n=4, mean±SEM, P

    Journal: Science signaling

    Article Title: Chemical synapses without synaptic vesicles: purinergic neurotransmission via a CALHM1 channel-mitochondrial signaling complex

    doi: 10.1126/scisignal.aao1815

    Figure Lengend Snippet: Blocking ATP production in mitochondria blocks release of ATP Oligomycin blocks ATP synthase in mitochondria allowing for the build-up of ADP within the matrix and intermembrane spaces. (A) ATP biosensor cells were transfected with mRNAs encoding P2X2/P2X3 ionotropic purinergic receptors, which are responsive to ATP but not ADP. (B) Before application of oligomycin, taste cells show a robust and repeatable release of ATP (blue trace). Five minutes after application of oligomycin, depolarization-evoked ATP release was eliminated (red trace). The bar graph shows a quantitative comparison of the responses (5 minutes with oligomycin, n=4, mean±SEM, P

    Article Snippet: Primary antibodies employed were: mouse monoclonal antibody 32C2 directed against the C-terminal domain of CALHM1 at 1:50 dilution ( ); rabbit polyclonal antibody against cytochrome C (Santa Cruz, sc-7159; AB_2090474); rabbit anti-P2X3 at 1:1000 dilution (APR-016; Alomone Labs; Jerusalem, Israel; AB_2341047); chicken anti GFP at 1:1000 dilution (Aves Labs; Tigard, OR; AB_10000240).

    Techniques: Blocking Assay, Transfection

    Go6976 reduces α,β-meATP–elicited flinch nocifensive responses in complete Freund adjuvant–treated rats, and PKCβ1 is not expressed in P2X3R-containing dorsal root ganglia (DRG) neurons. (A) In saline control rats, Go6976 (11.5 pmol = 4.35 ng in 50-μL saline) had no effect on α,β-meATP–elicited flinch activity. In complete Freund adjuvant–treated rats, Go6976 inhibited the enhanced α,β-meATP–induced flinch responses (n = 6; * P

    Journal: Pain

    Article Title: Epac–protein kinase C alpha signaling in purinergic P2X3R-mediated hyperalgesia after inflammation

    doi: 10.1097/j.pain.0000000000000547

    Figure Lengend Snippet: Go6976 reduces α,β-meATP–elicited flinch nocifensive responses in complete Freund adjuvant–treated rats, and PKCβ1 is not expressed in P2X3R-containing dorsal root ganglia (DRG) neurons. (A) In saline control rats, Go6976 (11.5 pmol = 4.35 ng in 50-μL saline) had no effect on α,β-meATP–elicited flinch activity. In complete Freund adjuvant–treated rats, Go6976 inhibited the enhanced α,β-meATP–induced flinch responses (n = 6; * P

    Article Snippet: Antibodies used were mouse anti-Epac1, anti-Epac2 (1:1000; Cell Signaling, Danvers, MA), mouse anti-PKCε (1:1000; Santa Cruz), rabbit anti-pPKCε (1:1000; Santa Cruz), and rabbit anti-P2X3R (1:2000; Alomone Labs). β-Actin, probed with anti-β-actin (1:10,000; Chemicon, Temeculla, CA), or the neuronal marker, β-tubulin, probed with anti β-tubulin antibody (1:2000; Santa Cruz), was used as loading control.

    Techniques: Activity Assay

    Phosphorylated PKC (pPKC) isoform and P2X3R expression in dorsal root ganglia slices prepared from complete Freund adjuvant–treated rats. Upper: pPKCα and P2X3Rs were expressed only in small or medium cells. pPKCα labels were found both at the cell membrane and in the cytoplasm. P2X3Rs were colocalized with pPKCα in small or medium cells. Enlarged views of cells (indicated by arrows) are shown in the lower left corners. Lower: pPKCε was expressed in both small or medium and large cells, most contained punctate labels. pPKCε labels were colocalized with P2X3Rs in small or medium cells (bar scale = 25 μm).

    Journal: Pain

    Article Title: Epac–protein kinase C alpha signaling in purinergic P2X3R-mediated hyperalgesia after inflammation

    doi: 10.1097/j.pain.0000000000000547

    Figure Lengend Snippet: Phosphorylated PKC (pPKC) isoform and P2X3R expression in dorsal root ganglia slices prepared from complete Freund adjuvant–treated rats. Upper: pPKCα and P2X3Rs were expressed only in small or medium cells. pPKCα labels were found both at the cell membrane and in the cytoplasm. P2X3Rs were colocalized with pPKCα in small or medium cells. Enlarged views of cells (indicated by arrows) are shown in the lower left corners. Lower: pPKCε was expressed in both small or medium and large cells, most contained punctate labels. pPKCε labels were colocalized with P2X3Rs in small or medium cells (bar scale = 25 μm).

    Article Snippet: Antibodies used were mouse anti-Epac1, anti-Epac2 (1:1000; Cell Signaling, Danvers, MA), mouse anti-PKCε (1:1000; Santa Cruz), rabbit anti-pPKCε (1:1000; Santa Cruz), and rabbit anti-P2X3R (1:2000; Alomone Labs). β-Actin, probed with anti-β-actin (1:10,000; Chemicon, Temeculla, CA), or the neuronal marker, β-tubulin, probed with anti β-tubulin antibody (1:2000; Santa Cruz), was used as loading control.

    Techniques: Expressing