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rabbit anti- pde2a, fabgennix, pde2a-101ap  (FabGennix)

 
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    FabGennix rabbit anti- pde2a, fabgennix, pde2a-101ap
    Rabbit Anti Pde2a, Fabgennix, Pde2a 101ap, supplied by FabGennix, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rabbit+anti-pde2a/rabbit+anti+pde10a+antibody/10__1161_slash_jaha__124__037343-170-56-55
    Average 90 stars, based on 1 article reviews
    rabbit anti- pde2a, fabgennix, pde2a-101ap - by Bioz Stars, 2026-10
    90/100 stars

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    Related Articles

    Immunostaining:

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat
    Article Snippet: *Rabbit anti-PDE2A; FabGennix; 1:400; #PD2A-101AP, lot FGI.159.Pld.Ig, Polyclonal.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators.
    Article Snippet: The mammalian olfactory system detects and discriminates between millions of odorants to elicit appropriate behavioral responses.. While much has been learned about how olfactory sensory neurons detect odorants and signal their presence, how specific innate, unlearned behaviors are initiated in response to ethologically relevant odors remains poorly understood.. Here, we show that the 4-transmembrane protein CD20, also known as MS4A1, is expressed in a previously uncharacterized subpopulation of olfactory sensory neurons in the main olfactory epithelium of the murine nasal cavity and functions as a mammalian olfactory receptor that recognizes compounds produced by mouse predators.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators
    Article Snippet: Primary antibodies/concentrations used were as follows: rabbit anti-phospho-S6 ribosomal protein (Serine240/244) (1:100, Cell Signaling Technologies, #2215), rabbit anti-phospho-S6 ribosomal protein (Serine244/247) (1:150, Invitrogen, #44-923G), rabbit anti-MS4A1/CD20 (1:250 for immunostaining, 1:100 for iDISCO, Cell Signaling Technology, #98708), rabbit anti-MS4A1 (1:200, MyBioSource, #MBS2051903), goat anti-MS4A1 (1:50, Santa Cruz Biotechnology, #sc-7735), rat anti-MS4A1 (1:100, LifeSpan Biosciences, #LS-C107163-100), guinea pig anti-VGLUT2 (1:500, SYSY, #135 404), rabbit anti-NeuN (1:500, Abcam, #ab104225), rabbit anti-KI18 (1:500, Abcam, #ab52948), rabbit anti-KI17 (1:500, Abcam, #ab53707), goat anti-NeuroD1 (1:50, R&D Systems, #AF2746), goat anti-OMP (1:1000, Wako Chemicals, #544-10001-WAKO), rabbit anti-CNGA2 (1:200, Alomone Labs, #APC-045), and rabbit anti-PDE2A (1:500, FabGennix, #PD2A-101AP).

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat.
    Article Snippet: Rabbit anti-PDE2A; FabGennix; 1:400) were applied overnight in 5% NGS.

    Expressing:

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat
    Article Snippet: *Rabbit anti-PDE2A; FabGennix; 1:400; #PD2A-101AP, lot FGI.159.Pld.Ig, Polyclonal.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators.
    Article Snippet: The mammalian olfactory system detects and discriminates between millions of odorants to elicit appropriate behavioral responses.. While much has been learned about how olfactory sensory neurons detect odorants and signal their presence, how specific innate, unlearned behaviors are initiated in response to ethologically relevant odors remains poorly understood.. Here, we show that the 4-transmembrane protein CD20, also known as MS4A1, is expressed in a previously uncharacterized subpopulation of olfactory sensory neurons in the main olfactory epithelium of the murine nasal cavity and functions as a mammalian olfactory receptor that recognizes compounds produced by mouse predators.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators
    Article Snippet: Primary antibodies/concentrations used were as follows: rabbit anti-phospho-S6 ribosomal protein (Serine240/244) (1:100, Cell Signaling Technologies, #2215), rabbit anti-phospho-S6 ribosomal protein (Serine244/247) (1:150, Invitrogen, #44-923G), rabbit anti-MS4A1/CD20 (1:250 for immunostaining, 1:100 for iDISCO, Cell Signaling Technology, #98708), rabbit anti-MS4A1 (1:200, MyBioSource, #MBS2051903), goat anti-MS4A1 (1:50, Santa Cruz Biotechnology, #sc-7735), rat anti-MS4A1 (1:100, LifeSpan Biosciences, #LS-C107163-100), guinea pig anti-VGLUT2 (1:500, SYSY, #135 404), rabbit anti-NeuN (1:500, Abcam, #ab104225), rabbit anti-KI18 (1:500, Abcam, #ab52948), rabbit anti-KI17 (1:500, Abcam, #ab53707), goat anti-NeuroD1 (1:50, R&D Systems, #AF2746), goat anti-OMP (1:1000, Wako Chemicals, #544-10001-WAKO), rabbit anti-CNGA2 (1:200, Alomone Labs, #APC-045), and rabbit anti-PDE2A (1:500, FabGennix, #PD2A-101AP).

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat.
    Article Snippet: Rabbit anti-PDE2A; FabGennix; 1:400) were applied overnight in 5% NGS.

    Knock-Out:

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat
    Article Snippet: *Rabbit anti-PDE2A; FabGennix; 1:400; #PD2A-101AP, lot FGI.159.Pld.Ig, Polyclonal.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators.
    Article Snippet: The mammalian olfactory system detects and discriminates between millions of odorants to elicit appropriate behavioral responses.. While much has been learned about how olfactory sensory neurons detect odorants and signal their presence, how specific innate, unlearned behaviors are initiated in response to ethologically relevant odors remains poorly understood.. Here, we show that the 4-transmembrane protein CD20, also known as MS4A1, is expressed in a previously uncharacterized subpopulation of olfactory sensory neurons in the main olfactory epithelium of the murine nasal cavity and functions as a mammalian olfactory receptor that recognizes compounds produced by mouse predators.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators
    Article Snippet: Primary antibodies/concentrations used were as follows: rabbit anti-phospho-S6 ribosomal protein (Serine240/244) (1:100, Cell Signaling Technologies, #2215), rabbit anti-phospho-S6 ribosomal protein (Serine244/247) (1:150, Invitrogen, #44-923G), rabbit anti-MS4A1/CD20 (1:250 for immunostaining, 1:100 for iDISCO, Cell Signaling Technology, #98708), rabbit anti-MS4A1 (1:200, MyBioSource, #MBS2051903), goat anti-MS4A1 (1:50, Santa Cruz Biotechnology, #sc-7735), rat anti-MS4A1 (1:100, LifeSpan Biosciences, #LS-C107163-100), guinea pig anti-VGLUT2 (1:500, SYSY, #135 404), rabbit anti-NeuN (1:500, Abcam, #ab104225), rabbit anti-KI18 (1:500, Abcam, #ab52948), rabbit anti-KI17 (1:500, Abcam, #ab53707), goat anti-NeuroD1 (1:50, R&D Systems, #AF2746), goat anti-OMP (1:1000, Wako Chemicals, #544-10001-WAKO), rabbit anti-CNGA2 (1:200, Alomone Labs, #APC-045), and rabbit anti-PDE2A (1:500, FabGennix, #PD2A-101AP).

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat.
    Article Snippet: Rabbit anti-PDE2A; FabGennix; 1:400) were applied overnight in 5% NGS.

    Labeling:

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat
    Article Snippet: *Rabbit anti-PDE2A; FabGennix; 1:400; #PD2A-101AP, lot FGI.159.Pld.Ig, Polyclonal.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators.
    Article Snippet: The mammalian olfactory system detects and discriminates between millions of odorants to elicit appropriate behavioral responses.. While much has been learned about how olfactory sensory neurons detect odorants and signal their presence, how specific innate, unlearned behaviors are initiated in response to ethologically relevant odors remains poorly understood.. Here, we show that the 4-transmembrane protein CD20, also known as MS4A1, is expressed in a previously uncharacterized subpopulation of olfactory sensory neurons in the main olfactory epithelium of the murine nasal cavity and functions as a mammalian olfactory receptor that recognizes compounds produced by mouse predators.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators
    Article Snippet: Primary antibodies/concentrations used were as follows: rabbit anti-phospho-S6 ribosomal protein (Serine240/244) (1:100, Cell Signaling Technologies, #2215), rabbit anti-phospho-S6 ribosomal protein (Serine244/247) (1:150, Invitrogen, #44-923G), rabbit anti-MS4A1/CD20 (1:250 for immunostaining, 1:100 for iDISCO, Cell Signaling Technology, #98708), rabbit anti-MS4A1 (1:200, MyBioSource, #MBS2051903), goat anti-MS4A1 (1:50, Santa Cruz Biotechnology, #sc-7735), rat anti-MS4A1 (1:100, LifeSpan Biosciences, #LS-C107163-100), guinea pig anti-VGLUT2 (1:500, SYSY, #135 404), rabbit anti-NeuN (1:500, Abcam, #ab104225), rabbit anti-KI18 (1:500, Abcam, #ab52948), rabbit anti-KI17 (1:500, Abcam, #ab53707), goat anti-NeuroD1 (1:50, R&D Systems, #AF2746), goat anti-OMP (1:1000, Wako Chemicals, #544-10001-WAKO), rabbit anti-CNGA2 (1:200, Alomone Labs, #APC-045), and rabbit anti-PDE2A (1:500, FabGennix, #PD2A-101AP).

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat.
    Article Snippet: Rabbit anti-PDE2A; FabGennix; 1:400) were applied overnight in 5% NGS.

    In Situ Hybridization:

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat
    Article Snippet: *Rabbit anti-PDE2A; FabGennix; 1:400; #PD2A-101AP, lot FGI.159.Pld.Ig, Polyclonal.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators.
    Article Snippet: The mammalian olfactory system detects and discriminates between millions of odorants to elicit appropriate behavioral responses.. While much has been learned about how olfactory sensory neurons detect odorants and signal their presence, how specific innate, unlearned behaviors are initiated in response to ethologically relevant odors remains poorly understood.. Here, we show that the 4-transmembrane protein CD20, also known as MS4A1, is expressed in a previously uncharacterized subpopulation of olfactory sensory neurons in the main olfactory epithelium of the murine nasal cavity and functions as a mammalian olfactory receptor that recognizes compounds produced by mouse predators.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators
    Article Snippet: Primary antibodies/concentrations used were as follows: rabbit anti-phospho-S6 ribosomal protein (Serine240/244) (1:100, Cell Signaling Technologies, #2215), rabbit anti-phospho-S6 ribosomal protein (Serine244/247) (1:150, Invitrogen, #44-923G), rabbit anti-MS4A1/CD20 (1:250 for immunostaining, 1:100 for iDISCO, Cell Signaling Technology, #98708), rabbit anti-MS4A1 (1:200, MyBioSource, #MBS2051903), goat anti-MS4A1 (1:50, Santa Cruz Biotechnology, #sc-7735), rat anti-MS4A1 (1:100, LifeSpan Biosciences, #LS-C107163-100), guinea pig anti-VGLUT2 (1:500, SYSY, #135 404), rabbit anti-NeuN (1:500, Abcam, #ab104225), rabbit anti-KI18 (1:500, Abcam, #ab52948), rabbit anti-KI17 (1:500, Abcam, #ab53707), goat anti-NeuroD1 (1:50, R&D Systems, #AF2746), goat anti-OMP (1:1000, Wako Chemicals, #544-10001-WAKO), rabbit anti-CNGA2 (1:200, Alomone Labs, #APC-045), and rabbit anti-PDE2A (1:500, FabGennix, #PD2A-101AP).

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat.
    Article Snippet: Rabbit anti-PDE2A; FabGennix; 1:400) were applied overnight in 5% NGS.

    Immunohistochemistry:

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat
    Article Snippet: *Rabbit anti-PDE2A; FabGennix; 1:400; #PD2A-101AP, lot FGI.159.Pld.Ig, Polyclonal.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators.
    Article Snippet: The mammalian olfactory system detects and discriminates between millions of odorants to elicit appropriate behavioral responses.. While much has been learned about how olfactory sensory neurons detect odorants and signal their presence, how specific innate, unlearned behaviors are initiated in response to ethologically relevant odors remains poorly understood.. Here, we show that the 4-transmembrane protein CD20, also known as MS4A1, is expressed in a previously uncharacterized subpopulation of olfactory sensory neurons in the main olfactory epithelium of the murine nasal cavity and functions as a mammalian olfactory receptor that recognizes compounds produced by mouse predators.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators
    Article Snippet: Primary antibodies/concentrations used were as follows: rabbit anti-phospho-S6 ribosomal protein (Serine240/244) (1:100, Cell Signaling Technologies, #2215), rabbit anti-phospho-S6 ribosomal protein (Serine244/247) (1:150, Invitrogen, #44-923G), rabbit anti-MS4A1/CD20 (1:250 for immunostaining, 1:100 for iDISCO, Cell Signaling Technology, #98708), rabbit anti-MS4A1 (1:200, MyBioSource, #MBS2051903), goat anti-MS4A1 (1:50, Santa Cruz Biotechnology, #sc-7735), rat anti-MS4A1 (1:100, LifeSpan Biosciences, #LS-C107163-100), guinea pig anti-VGLUT2 (1:500, SYSY, #135 404), rabbit anti-NeuN (1:500, Abcam, #ab104225), rabbit anti-KI18 (1:500, Abcam, #ab52948), rabbit anti-KI17 (1:500, Abcam, #ab53707), goat anti-NeuroD1 (1:50, R&D Systems, #AF2746), goat anti-OMP (1:1000, Wako Chemicals, #544-10001-WAKO), rabbit anti-CNGA2 (1:200, Alomone Labs, #APC-045), and rabbit anti-PDE2A (1:500, FabGennix, #PD2A-101AP).

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat.
    Article Snippet: Rabbit anti-PDE2A; FabGennix; 1:400) were applied overnight in 5% NGS.

    Marker:

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat
    Article Snippet: *Rabbit anti-PDE2A; FabGennix; 1:400; #PD2A-101AP, lot FGI.159.Pld.Ig, Polyclonal.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators.
    Article Snippet: The mammalian olfactory system detects and discriminates between millions of odorants to elicit appropriate behavioral responses.. While much has been learned about how olfactory sensory neurons detect odorants and signal their presence, how specific innate, unlearned behaviors are initiated in response to ethologically relevant odors remains poorly understood.. Here, we show that the 4-transmembrane protein CD20, also known as MS4A1, is expressed in a previously uncharacterized subpopulation of olfactory sensory neurons in the main olfactory epithelium of the murine nasal cavity and functions as a mammalian olfactory receptor that recognizes compounds produced by mouse predators.

    Article Title: CD20/MS4A1 is a mammalian olfactory receptor expressed in a subset of olfactory sensory neurons that mediates innate avoidance of predators
    Article Snippet: Primary antibodies/concentrations used were as follows: rabbit anti-phospho-S6 ribosomal protein (Serine240/244) (1:100, Cell Signaling Technologies, #2215), rabbit anti-phospho-S6 ribosomal protein (Serine244/247) (1:150, Invitrogen, #44-923G), rabbit anti-MS4A1/CD20 (1:250 for immunostaining, 1:100 for iDISCO, Cell Signaling Technology, #98708), rabbit anti-MS4A1 (1:200, MyBioSource, #MBS2051903), goat anti-MS4A1 (1:50, Santa Cruz Biotechnology, #sc-7735), rat anti-MS4A1 (1:100, LifeSpan Biosciences, #LS-C107163-100), guinea pig anti-VGLUT2 (1:500, SYSY, #135 404), rabbit anti-NeuN (1:500, Abcam, #ab104225), rabbit anti-KI18 (1:500, Abcam, #ab52948), rabbit anti-KI17 (1:500, Abcam, #ab53707), goat anti-NeuroD1 (1:50, R&D Systems, #AF2746), goat anti-OMP (1:1000, Wako Chemicals, #544-10001-WAKO), rabbit anti-CNGA2 (1:200, Alomone Labs, #APC-045), and rabbit anti-PDE2A (1:500, FabGennix, #PD2A-101AP).

    Article Title: The Grueneberg ganglion controls odor-driven food choices in mice under threat.
    Article Snippet: Rabbit anti-PDE2A; FabGennix; 1:400) were applied overnight in 5% NGS.



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    Figure 2. Gene therapy with <t>PDE2A</t> prevents cardiac remodeling and dysfunction induced by chronic isoproterenol infusion. A, Schematic representation of the experimental protocol. Mice were injected in the tail vein with 1012 viral particles of a serotype 9 adeno-associated viruses encoding for luciferase or phosphodiesterase 2A3. Fourteen days later, mice injected with AAV9-LUC were implanted subcutaneously with osmotic pumps diffusing either NaCl (NaCl-LUC) or isoproterenol at 60 mg/kg per day (isoproterenol- LUC) and animals injected with AAV9-PDE2A were implanted with pumps delivering isoproterenol at 60 mg/kg per day (isoproterenol- PDE2A). Treatment duration was 14 days (hatched bars). Minipumps were then removed and mice were kept for 2 additional weeks before euthanasia. Cardiac function was evaluated by serial echocardiography before injection of the virus as well as at 14, 28, 35, and 42 days. B, Left panel shows representative blots of PDE2A and vinculin. PDE2A protein expression in heart tissues extracts measured by Western blot and the bar graph represents the ratios of PDE2A over vinculin quantified expressed as mean±SEM in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A groups. C, Heart weight and lung weight over tibia length ratio expressed as mean±SEM. D, Time course of the normalized ratio of calculated left ventricular weight over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A mice. Number of mice is indicated in the brackets. E, Panel on the left, representative images of Masson’s trichrome staining (scale bar 100 μm), bar graph on the left represents quantifications of interstitial fibrosis as mean±SEM in NaCl-LUC (n=9), isoproterenol-LUC (n=9), and isoproterenol+PDE2A (n=9) groups. Statistical significance is indicated by the P value (* vs NaCl-LUC and † vs isoproterenol-LUC) determined using Welch’s t test (B), Kruskal– Wallis followed by a Dunn’s test (C panel bottom and E), 2-way ANOVA followed by a Tukey’s test (D), 1-way ANOVA followed by a Tukey’s test (C panel top). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-LUC, isoproterenol-luciferase; Iso-PDE2A, isoproterenol-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; PDE2A, phosphodiesterase 2A; and TL, tibia length.
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    FabGennix rabbit anti-pde2a fabgennix
    Figure 2. Gene therapy with <t>PDE2A</t> prevents cardiac remodeling and dysfunction induced by chronic isoproterenol infusion. A, Schematic representation of the experimental protocol. Mice were injected in the tail vein with 1012 viral particles of a serotype 9 adeno-associated viruses encoding for luciferase or phosphodiesterase 2A3. Fourteen days later, mice injected with AAV9-LUC were implanted subcutaneously with osmotic pumps diffusing either NaCl (NaCl-LUC) or isoproterenol at 60 mg/kg per day (isoproterenol- LUC) and animals injected with AAV9-PDE2A were implanted with pumps delivering isoproterenol at 60 mg/kg per day (isoproterenol- PDE2A). Treatment duration was 14 days (hatched bars). Minipumps were then removed and mice were kept for 2 additional weeks before euthanasia. Cardiac function was evaluated by serial echocardiography before injection of the virus as well as at 14, 28, 35, and 42 days. B, Left panel shows representative blots of PDE2A and vinculin. PDE2A protein expression in heart tissues extracts measured by Western blot and the bar graph represents the ratios of PDE2A over vinculin quantified expressed as mean±SEM in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A groups. C, Heart weight and lung weight over tibia length ratio expressed as mean±SEM. D, Time course of the normalized ratio of calculated left ventricular weight over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A mice. Number of mice is indicated in the brackets. E, Panel on the left, representative images of Masson’s trichrome staining (scale bar 100 μm), bar graph on the left represents quantifications of interstitial fibrosis as mean±SEM in NaCl-LUC (n=9), isoproterenol-LUC (n=9), and isoproterenol+PDE2A (n=9) groups. Statistical significance is indicated by the P value (* vs NaCl-LUC and † vs isoproterenol-LUC) determined using Welch’s t test (B), Kruskal– Wallis followed by a Dunn’s test (C panel bottom and E), 2-way ANOVA followed by a Tukey’s test (D), 1-way ANOVA followed by a Tukey’s test (C panel top). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-LUC, isoproterenol-luciferase; Iso-PDE2A, isoproterenol-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; PDE2A, phosphodiesterase 2A; and TL, tibia length.
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    Figure 2. Gene therapy with <t>PDE2A</t> prevents cardiac remodeling and dysfunction induced by chronic isoproterenol infusion. A, Schematic representation of the experimental protocol. Mice were injected in the tail vein with 1012 viral particles of a serotype 9 adeno-associated viruses encoding for luciferase or phosphodiesterase 2A3. Fourteen days later, mice injected with AAV9-LUC were implanted subcutaneously with osmotic pumps diffusing either NaCl (NaCl-LUC) or isoproterenol at 60 mg/kg per day (isoproterenol- LUC) and animals injected with AAV9-PDE2A were implanted with pumps delivering isoproterenol at 60 mg/kg per day (isoproterenol- PDE2A). Treatment duration was 14 days (hatched bars). Minipumps were then removed and mice were kept for 2 additional weeks before euthanasia. Cardiac function was evaluated by serial echocardiography before injection of the virus as well as at 14, 28, 35, and 42 days. B, Left panel shows representative blots of PDE2A and vinculin. PDE2A protein expression in heart tissues extracts measured by Western blot and the bar graph represents the ratios of PDE2A over vinculin quantified expressed as mean±SEM in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A groups. C, Heart weight and lung weight over tibia length ratio expressed as mean±SEM. D, Time course of the normalized ratio of calculated left ventricular weight over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A mice. Number of mice is indicated in the brackets. E, Panel on the left, representative images of Masson’s trichrome staining (scale bar 100 μm), bar graph on the left represents quantifications of interstitial fibrosis as mean±SEM in NaCl-LUC (n=9), isoproterenol-LUC (n=9), and isoproterenol+PDE2A (n=9) groups. Statistical significance is indicated by the P value (* vs NaCl-LUC and † vs isoproterenol-LUC) determined using Welch’s t test (B), Kruskal– Wallis followed by a Dunn’s test (C panel bottom and E), 2-way ANOVA followed by a Tukey’s test (D), 1-way ANOVA followed by a Tukey’s test (C panel top). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-LUC, isoproterenol-luciferase; Iso-PDE2A, isoproterenol-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; PDE2A, phosphodiesterase 2A; and TL, tibia length.
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    Figure 2. Gene therapy with <t>PDE2A</t> prevents cardiac remodeling and dysfunction induced by chronic isoproterenol infusion. A, Schematic representation of the experimental protocol. Mice were injected in the tail vein with 1012 viral particles of a serotype 9 adeno-associated viruses encoding for luciferase or phosphodiesterase 2A3. Fourteen days later, mice injected with AAV9-LUC were implanted subcutaneously with osmotic pumps diffusing either NaCl (NaCl-LUC) or isoproterenol at 60 mg/kg per day (isoproterenol- LUC) and animals injected with AAV9-PDE2A were implanted with pumps delivering isoproterenol at 60 mg/kg per day (isoproterenol- PDE2A). Treatment duration was 14 days (hatched bars). Minipumps were then removed and mice were kept for 2 additional weeks before euthanasia. Cardiac function was evaluated by serial echocardiography before injection of the virus as well as at 14, 28, 35, and 42 days. B, Left panel shows representative blots of PDE2A and vinculin. PDE2A protein expression in heart tissues extracts measured by Western blot and the bar graph represents the ratios of PDE2A over vinculin quantified expressed as mean±SEM in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A groups. C, Heart weight and lung weight over tibia length ratio expressed as mean±SEM. D, Time course of the normalized ratio of calculated left ventricular weight over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A mice. Number of mice is indicated in the brackets. E, Panel on the left, representative images of Masson’s trichrome staining (scale bar 100 μm), bar graph on the left represents quantifications of interstitial fibrosis as mean±SEM in NaCl-LUC (n=9), isoproterenol-LUC (n=9), and isoproterenol+PDE2A (n=9) groups. Statistical significance is indicated by the P value (* vs NaCl-LUC and † vs isoproterenol-LUC) determined using Welch’s t test (B), Kruskal– Wallis followed by a Dunn’s test (C panel bottom and E), 2-way ANOVA followed by a Tukey’s test (D), 1-way ANOVA followed by a Tukey’s test (C panel top). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-LUC, isoproterenol-luciferase; Iso-PDE2A, isoproterenol-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; PDE2A, phosphodiesterase 2A; and TL, tibia length.
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    Figure 2. Gene therapy with <t>PDE2A</t> prevents cardiac remodeling and dysfunction induced by chronic isoproterenol infusion. A, Schematic representation of the experimental protocol. Mice were injected in the tail vein with 1012 viral particles of a serotype 9 adeno-associated viruses encoding for luciferase or phosphodiesterase 2A3. Fourteen days later, mice injected with AAV9-LUC were implanted subcutaneously with osmotic pumps diffusing either NaCl (NaCl-LUC) or isoproterenol at 60 mg/kg per day (isoproterenol- LUC) and animals injected with AAV9-PDE2A were implanted with pumps delivering isoproterenol at 60 mg/kg per day (isoproterenol- PDE2A). Treatment duration was 14 days (hatched bars). Minipumps were then removed and mice were kept for 2 additional weeks before euthanasia. Cardiac function was evaluated by serial echocardiography before injection of the virus as well as at 14, 28, 35, and 42 days. B, Left panel shows representative blots of PDE2A and vinculin. PDE2A protein expression in heart tissues extracts measured by Western blot and the bar graph represents the ratios of PDE2A over vinculin quantified expressed as mean±SEM in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A groups. C, Heart weight and lung weight over tibia length ratio expressed as mean±SEM. D, Time course of the normalized ratio of calculated left ventricular weight over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A mice. Number of mice is indicated in the brackets. E, Panel on the left, representative images of Masson’s trichrome staining (scale bar 100 μm), bar graph on the left represents quantifications of interstitial fibrosis as mean±SEM in NaCl-LUC (n=9), isoproterenol-LUC (n=9), and isoproterenol+PDE2A (n=9) groups. Statistical significance is indicated by the P value (* vs NaCl-LUC and † vs isoproterenol-LUC) determined using Welch’s t test (B), Kruskal– Wallis followed by a Dunn’s test (C panel bottom and E), 2-way ANOVA followed by a Tukey’s test (D), 1-way ANOVA followed by a Tukey’s test (C panel top). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-LUC, isoproterenol-luciferase; Iso-PDE2A, isoproterenol-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; PDE2A, phosphodiesterase 2A; and TL, tibia length.
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    Figure 2. Gene therapy with <t>PDE2A</t> prevents cardiac remodeling and dysfunction induced by chronic isoproterenol infusion. A, Schematic representation of the experimental protocol. Mice were injected in the tail vein with 1012 viral particles of a serotype 9 adeno-associated viruses encoding for luciferase or phosphodiesterase 2A3. Fourteen days later, mice injected with AAV9-LUC were implanted subcutaneously with osmotic pumps diffusing either NaCl (NaCl-LUC) or isoproterenol at 60 mg/kg per day (isoproterenol- LUC) and animals injected with AAV9-PDE2A were implanted with pumps delivering isoproterenol at 60 mg/kg per day (isoproterenol- PDE2A). Treatment duration was 14 days (hatched bars). Minipumps were then removed and mice were kept for 2 additional weeks before euthanasia. Cardiac function was evaluated by serial echocardiography before injection of the virus as well as at 14, 28, 35, and 42 days. B, Left panel shows representative blots of PDE2A and vinculin. PDE2A protein expression in heart tissues extracts measured by Western blot and the bar graph represents the ratios of PDE2A over vinculin quantified expressed as mean±SEM in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A groups. C, Heart weight and lung weight over tibia length ratio expressed as mean±SEM. D, Time course of the normalized ratio of calculated left ventricular weight over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A mice. Number of mice is indicated in the brackets. E, Panel on the left, representative images of Masson’s trichrome staining (scale bar 100 μm), bar graph on the left represents quantifications of interstitial fibrosis as mean±SEM in NaCl-LUC (n=9), isoproterenol-LUC (n=9), and isoproterenol+PDE2A (n=9) groups. Statistical significance is indicated by the P value (* vs NaCl-LUC and † vs isoproterenol-LUC) determined using Welch’s t test (B), Kruskal– Wallis followed by a Dunn’s test (C panel bottom and E), 2-way ANOVA followed by a Tukey’s test (D), 1-way ANOVA followed by a Tukey’s test (C panel top). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-LUC, isoproterenol-luciferase; Iso-PDE2A, isoproterenol-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; PDE2A, phosphodiesterase 2A; and TL, tibia length.
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    Figure 2. Gene therapy with PDE2A prevents cardiac remodeling and dysfunction induced by chronic isoproterenol infusion. A, Schematic representation of the experimental protocol. Mice were injected in the tail vein with 1012 viral particles of a serotype 9 adeno-associated viruses encoding for luciferase or phosphodiesterase 2A3. Fourteen days later, mice injected with AAV9-LUC were implanted subcutaneously with osmotic pumps diffusing either NaCl (NaCl-LUC) or isoproterenol at 60 mg/kg per day (isoproterenol- LUC) and animals injected with AAV9-PDE2A were implanted with pumps delivering isoproterenol at 60 mg/kg per day (isoproterenol- PDE2A). Treatment duration was 14 days (hatched bars). Minipumps were then removed and mice were kept for 2 additional weeks before euthanasia. Cardiac function was evaluated by serial echocardiography before injection of the virus as well as at 14, 28, 35, and 42 days. B, Left panel shows representative blots of PDE2A and vinculin. PDE2A protein expression in heart tissues extracts measured by Western blot and the bar graph represents the ratios of PDE2A over vinculin quantified expressed as mean±SEM in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A groups. C, Heart weight and lung weight over tibia length ratio expressed as mean±SEM. D, Time course of the normalized ratio of calculated left ventricular weight over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A mice. Number of mice is indicated in the brackets. E, Panel on the left, representative images of Masson’s trichrome staining (scale bar 100 μm), bar graph on the left represents quantifications of interstitial fibrosis as mean±SEM in NaCl-LUC (n=9), isoproterenol-LUC (n=9), and isoproterenol+PDE2A (n=9) groups. Statistical significance is indicated by the P value (* vs NaCl-LUC and † vs isoproterenol-LUC) determined using Welch’s t test (B), Kruskal– Wallis followed by a Dunn’s test (C panel bottom and E), 2-way ANOVA followed by a Tukey’s test (D), 1-way ANOVA followed by a Tukey’s test (C panel top). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-LUC, isoproterenol-luciferase; Iso-PDE2A, isoproterenol-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; PDE2A, phosphodiesterase 2A; and TL, tibia length.

    Journal: Journal of the American Heart Association

    Article Title: Cardiac Gene Therapy With Phosphodiesterase 2A Limits Remodeling and Arrhythmias in Mouse Models of Heart Failure

    doi: 10.1161/jaha.124.037343

    Figure Lengend Snippet: Figure 2. Gene therapy with PDE2A prevents cardiac remodeling and dysfunction induced by chronic isoproterenol infusion. A, Schematic representation of the experimental protocol. Mice were injected in the tail vein with 1012 viral particles of a serotype 9 adeno-associated viruses encoding for luciferase or phosphodiesterase 2A3. Fourteen days later, mice injected with AAV9-LUC were implanted subcutaneously with osmotic pumps diffusing either NaCl (NaCl-LUC) or isoproterenol at 60 mg/kg per day (isoproterenol- LUC) and animals injected with AAV9-PDE2A were implanted with pumps delivering isoproterenol at 60 mg/kg per day (isoproterenol- PDE2A). Treatment duration was 14 days (hatched bars). Minipumps were then removed and mice were kept for 2 additional weeks before euthanasia. Cardiac function was evaluated by serial echocardiography before injection of the virus as well as at 14, 28, 35, and 42 days. B, Left panel shows representative blots of PDE2A and vinculin. PDE2A protein expression in heart tissues extracts measured by Western blot and the bar graph represents the ratios of PDE2A over vinculin quantified expressed as mean±SEM in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A groups. C, Heart weight and lung weight over tibia length ratio expressed as mean±SEM. D, Time course of the normalized ratio of calculated left ventricular weight over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A mice. Number of mice is indicated in the brackets. E, Panel on the left, representative images of Masson’s trichrome staining (scale bar 100 μm), bar graph on the left represents quantifications of interstitial fibrosis as mean±SEM in NaCl-LUC (n=9), isoproterenol-LUC (n=9), and isoproterenol+PDE2A (n=9) groups. Statistical significance is indicated by the P value (* vs NaCl-LUC and † vs isoproterenol-LUC) determined using Welch’s t test (B), Kruskal– Wallis followed by a Dunn’s test (C panel bottom and E), 2-way ANOVA followed by a Tukey’s test (D), 1-way ANOVA followed by a Tukey’s test (C panel top). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-LUC, isoproterenol-luciferase; Iso-PDE2A, isoproterenol-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; PDE2A, phosphodiesterase 2A; and TL, tibia length.

    Article Snippet: The primary antibodies used were as follows: (1) Figure 1: rabbit anti- PDE2A, Fabgennix; PDE2A- 101AP diluted at 1/1000; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; (2) Figure 2: goat antiPDE2A, Santa Cruz, SC- 17228 diluted at 1/1000; mouse anti- vinculin, V9131 Sigma- Aldrich diluted at 1/1000; (3) Figure 3: rabbit anti- PDE2A, Fabgennix, PDE2A- 101AP diluted at 1/1000; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; (4) Figure S1: rabbit anti- flag L5, Novus Biologicals, NBP1- 06712 diluted at 1/500; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; rabbit anti- PDE2A, Proteintech, 55 306- 1- AP diluted at 1/1000; rabbit anti- flag L5, Novus Biologicals, NBP1- 06712 diluted at 1/500; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; (5) Figure S2B: rabbit anti- PDE2A, Proteintech, 55 306- 1- AP diluted at 1/1000; (6) Figure S3: rabbit antiPDE2A, Proteintech, 55 306- 1- AP diluted at 1/1000; and (7) Figure S4: rabbit anti- PDE4A AC55 (gift from Dr. Marco Conti, University of California San Francisco, San Francisco, CA) diluted at 1/1000; rabbit anti- PDE4B (gift from Dr. Marco Conti) diluted at 1/10000; mouse antiPDE4D (gift from Dr Marco Conti) diluted at 1/1000 and rabbit anti- PDE3A (gift from Dr Chen Yan, University of Rochester, Rochester, NY) diluted at 1/10000.

    Techniques: Injection, Luciferase, Virus, Expressing, Western Blot, Staining

    Figure 3. PDE2A overexpression limits maladaptive remodeling and cardiac dysfunction induced by chronic isoproterenol and phenylephrine infusion. A, Schematic representation of the experimental protocol. Mice were injected with 1012 viral particles of serotype 9 adeno-associated viruses encoding for Luciferase or phosphodiesterase 2A3. Fourteen days later, mice were implanted with osmotic minipumps diffusing either NaCl (NaCl-LUC) or isoproterenol+phenylephrine at 30 mg/kg per day each (isoproterenol+phenylephrine-LUC and isoproterenol + phenylephrine-PDE2A). Minipumps were removed 2 weeks later and mice were euthanized. Cardiac function was assessed throughout the protocol using echocardiography before the injection of the virus and at 2, 3, 4, and 6 weeks. B, Representative blots of PDE2A and GAPDH on the left panel are shown and the bar graph represents ratios of PDE2A over GAPDH quantifications expressed as mean±SEM in a subset of the animals treated with NaCl-LUC, isoproterenol+phenylephrine-LUC, and isoproterenol+phenylephrine-PDE2A. C, Heart weight and lung weight over tibia length ratios expressed as mean±SEM. D, Time course of the normalized ratios of calculated left ventricular weight (over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol+phenylephrine-LUC, and isoproterenol+phenylephrine-PDE2A mice). Number of mice is indicated in the brackets. E, Top panel shows representative images of Masson’s trichrome staining (scale bar 200 μm), the middle panel depicts representative images of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay (scale bar 100 μm) detecting apoptotic nuclei in green costained with the glycocalix marker wheat germ agglutinin in red and nuclei in blue colored with Hoechst. Bar graphs representing mean±SEM quantifications of interstitial fibrosis and apoptotic nuclei in NaCl-LUC (10), isoproterenol+phenylephrine- LUC (9) and isoproterenol+phenylephrine-PDE2A (9) mice are depicted. Statistical significance indicated by the P value (* vs NaCl- LUC and † vs isoproterenol+phenylephrine-LUC) was determined using Welch’s t test (B), Kruskal–Wallis followed by a Dunn’s test (C), 2-way ANOVA followed by a Tukey’s test (D), Welch’s 1-way ANOVA followed by a Dunnett’s test (E). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-Phe-LUC, isoproterenol-phenylephrine-luciferase; Iso-Phe- PDE2A, isoproterenol-phenylephrine-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; ns, not significant; PDE2A, phosphodiesterase 2A; and TL, tibia length.

    Journal: Journal of the American Heart Association

    Article Title: Cardiac Gene Therapy With Phosphodiesterase 2A Limits Remodeling and Arrhythmias in Mouse Models of Heart Failure

    doi: 10.1161/jaha.124.037343

    Figure Lengend Snippet: Figure 3. PDE2A overexpression limits maladaptive remodeling and cardiac dysfunction induced by chronic isoproterenol and phenylephrine infusion. A, Schematic representation of the experimental protocol. Mice were injected with 1012 viral particles of serotype 9 adeno-associated viruses encoding for Luciferase or phosphodiesterase 2A3. Fourteen days later, mice were implanted with osmotic minipumps diffusing either NaCl (NaCl-LUC) or isoproterenol+phenylephrine at 30 mg/kg per day each (isoproterenol+phenylephrine-LUC and isoproterenol + phenylephrine-PDE2A). Minipumps were removed 2 weeks later and mice were euthanized. Cardiac function was assessed throughout the protocol using echocardiography before the injection of the virus and at 2, 3, 4, and 6 weeks. B, Representative blots of PDE2A and GAPDH on the left panel are shown and the bar graph represents ratios of PDE2A over GAPDH quantifications expressed as mean±SEM in a subset of the animals treated with NaCl-LUC, isoproterenol+phenylephrine-LUC, and isoproterenol+phenylephrine-PDE2A. C, Heart weight and lung weight over tibia length ratios expressed as mean±SEM. D, Time course of the normalized ratios of calculated left ventricular weight (over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol+phenylephrine-LUC, and isoproterenol+phenylephrine-PDE2A mice). Number of mice is indicated in the brackets. E, Top panel shows representative images of Masson’s trichrome staining (scale bar 200 μm), the middle panel depicts representative images of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay (scale bar 100 μm) detecting apoptotic nuclei in green costained with the glycocalix marker wheat germ agglutinin in red and nuclei in blue colored with Hoechst. Bar graphs representing mean±SEM quantifications of interstitial fibrosis and apoptotic nuclei in NaCl-LUC (10), isoproterenol+phenylephrine- LUC (9) and isoproterenol+phenylephrine-PDE2A (9) mice are depicted. Statistical significance indicated by the P value (* vs NaCl- LUC and † vs isoproterenol+phenylephrine-LUC) was determined using Welch’s t test (B), Kruskal–Wallis followed by a Dunn’s test (C), 2-way ANOVA followed by a Tukey’s test (D), Welch’s 1-way ANOVA followed by a Dunnett’s test (E). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-Phe-LUC, isoproterenol-phenylephrine-luciferase; Iso-Phe- PDE2A, isoproterenol-phenylephrine-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; ns, not significant; PDE2A, phosphodiesterase 2A; and TL, tibia length.

    Article Snippet: The primary antibodies used were as follows: (1) Figure 1: rabbit anti- PDE2A, Fabgennix; PDE2A- 101AP diluted at 1/1000; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; (2) Figure 2: goat antiPDE2A, Santa Cruz, SC- 17228 diluted at 1/1000; mouse anti- vinculin, V9131 Sigma- Aldrich diluted at 1/1000; (3) Figure 3: rabbit anti- PDE2A, Fabgennix, PDE2A- 101AP diluted at 1/1000; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; (4) Figure S1: rabbit anti- flag L5, Novus Biologicals, NBP1- 06712 diluted at 1/500; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; rabbit anti- PDE2A, Proteintech, 55 306- 1- AP diluted at 1/1000; rabbit anti- flag L5, Novus Biologicals, NBP1- 06712 diluted at 1/500; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; (5) Figure S2B: rabbit anti- PDE2A, Proteintech, 55 306- 1- AP diluted at 1/1000; (6) Figure S3: rabbit antiPDE2A, Proteintech, 55 306- 1- AP diluted at 1/1000; and (7) Figure S4: rabbit anti- PDE4A AC55 (gift from Dr. Marco Conti, University of California San Francisco, San Francisco, CA) diluted at 1/1000; rabbit anti- PDE4B (gift from Dr. Marco Conti) diluted at 1/10000; mouse antiPDE4D (gift from Dr Marco Conti) diluted at 1/1000 and rabbit anti- PDE3A (gift from Dr Chen Yan, University of Rochester, Rochester, NY) diluted at 1/10000.

    Techniques: Over Expression, Injection, Luciferase, Virus, Staining, TUNEL Assay, Marker

    Figure 2. Gene therapy with PDE2A prevents cardiac remodeling and dysfunction induced by chronic isoproterenol infusion. A, Schematic representation of the experimental protocol. Mice were injected in the tail vein with 1012 viral particles of a serotype 9 adeno-associated viruses encoding for luciferase or phosphodiesterase 2A3. Fourteen days later, mice injected with AAV9-LUC were implanted subcutaneously with osmotic pumps diffusing either NaCl (NaCl-LUC) or isoproterenol at 60 mg/kg per day (isoproterenol- LUC) and animals injected with AAV9-PDE2A were implanted with pumps delivering isoproterenol at 60 mg/kg per day (isoproterenol- PDE2A). Treatment duration was 14 days (hatched bars). Minipumps were then removed and mice were kept for 2 additional weeks before euthanasia. Cardiac function was evaluated by serial echocardiography before injection of the virus as well as at 14, 28, 35, and 42 days. B, Left panel shows representative blots of PDE2A and vinculin. PDE2A protein expression in heart tissues extracts measured by Western blot and the bar graph represents the ratios of PDE2A over vinculin quantified expressed as mean±SEM in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A groups. C, Heart weight and lung weight over tibia length ratio expressed as mean±SEM. D, Time course of the normalized ratio of calculated left ventricular weight over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A mice. Number of mice is indicated in the brackets. E, Panel on the left, representative images of Masson’s trichrome staining (scale bar 100 μm), bar graph on the left represents quantifications of interstitial fibrosis as mean±SEM in NaCl-LUC (n=9), isoproterenol-LUC (n=9), and isoproterenol+PDE2A (n=9) groups. Statistical significance is indicated by the P value (* vs NaCl-LUC and † vs isoproterenol-LUC) determined using Welch’s t test (B), Kruskal– Wallis followed by a Dunn’s test (C panel bottom and E), 2-way ANOVA followed by a Tukey’s test (D), 1-way ANOVA followed by a Tukey’s test (C panel top). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-LUC, isoproterenol-luciferase; Iso-PDE2A, isoproterenol-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; PDE2A, phosphodiesterase 2A; and TL, tibia length.

    Journal: Journal of the American Heart Association

    Article Title: Cardiac Gene Therapy With Phosphodiesterase 2A Limits Remodeling and Arrhythmias in Mouse Models of Heart Failure

    doi: 10.1161/jaha.124.037343

    Figure Lengend Snippet: Figure 2. Gene therapy with PDE2A prevents cardiac remodeling and dysfunction induced by chronic isoproterenol infusion. A, Schematic representation of the experimental protocol. Mice were injected in the tail vein with 1012 viral particles of a serotype 9 adeno-associated viruses encoding for luciferase or phosphodiesterase 2A3. Fourteen days later, mice injected with AAV9-LUC were implanted subcutaneously with osmotic pumps diffusing either NaCl (NaCl-LUC) or isoproterenol at 60 mg/kg per day (isoproterenol- LUC) and animals injected with AAV9-PDE2A were implanted with pumps delivering isoproterenol at 60 mg/kg per day (isoproterenol- PDE2A). Treatment duration was 14 days (hatched bars). Minipumps were then removed and mice were kept for 2 additional weeks before euthanasia. Cardiac function was evaluated by serial echocardiography before injection of the virus as well as at 14, 28, 35, and 42 days. B, Left panel shows representative blots of PDE2A and vinculin. PDE2A protein expression in heart tissues extracts measured by Western blot and the bar graph represents the ratios of PDE2A over vinculin quantified expressed as mean±SEM in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A groups. C, Heart weight and lung weight over tibia length ratio expressed as mean±SEM. D, Time course of the normalized ratio of calculated left ventricular weight over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol-LUC and isoproterenol-PDE2A mice. Number of mice is indicated in the brackets. E, Panel on the left, representative images of Masson’s trichrome staining (scale bar 100 μm), bar graph on the left represents quantifications of interstitial fibrosis as mean±SEM in NaCl-LUC (n=9), isoproterenol-LUC (n=9), and isoproterenol+PDE2A (n=9) groups. Statistical significance is indicated by the P value (* vs NaCl-LUC and † vs isoproterenol-LUC) determined using Welch’s t test (B), Kruskal– Wallis followed by a Dunn’s test (C panel bottom and E), 2-way ANOVA followed by a Tukey’s test (D), 1-way ANOVA followed by a Tukey’s test (C panel top). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-LUC, isoproterenol-luciferase; Iso-PDE2A, isoproterenol-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; PDE2A, phosphodiesterase 2A; and TL, tibia length.

    Article Snippet: The primary antibodies used were as follows: (1) Figure 1: rabbit anti- PDE2A, Fabgennix; PDE2A- 101AP diluted at 1/1000; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; (2) Figure 2: goat antiPDE2A, Santa Cruz, SC- 17228 diluted at 1/1000; mouse anti- vinculin, V9131 Sigma- Aldrich diluted at 1/1000; (3) Figure 3: rabbit anti- PDE2A, Fabgennix, PDE2A- 101AP diluted at 1/1000; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; (4) Figure S1: rabbit anti- flag L5, Novus Biologicals, NBP1- 06712 diluted at 1/500; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; rabbit anti- PDE2A, Proteintech, 55 306- 1- AP diluted at 1/1000; rabbit anti- flag L5, Novus Biologicals, NBP1- 06712 diluted at 1/500; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; (5) Figure S2B: rabbit anti- PDE2A, Proteintech, 55 306- 1- AP diluted at 1/1000; (6) Figure S3: rabbit antiPDE2A, Proteintech, 55 306- 1- AP diluted at 1/1000; and (7) Figure S4: rabbit anti- PDE4A AC55 (gift from Dr. Marco Conti, University of California San Francisco, San Francisco, CA) diluted at 1/1000; rabbit anti- PDE4B (gift from Dr. Marco Conti) diluted at 1/10000; mouse antiPDE4D (gift from Dr Marco Conti) diluted at 1/1000 and rabbit anti- PDE3A (gift from Dr Chen Yan, University of Rochester, Rochester, NY) diluted at 1/10000.

    Techniques: Injection, Luciferase, Virus, Expressing, Western Blot, Staining

    Figure 3. PDE2A overexpression limits maladaptive remodeling and cardiac dysfunction induced by chronic isoproterenol and phenylephrine infusion. A, Schematic representation of the experimental protocol. Mice were injected with 1012 viral particles of serotype 9 adeno-associated viruses encoding for Luciferase or phosphodiesterase 2A3. Fourteen days later, mice were implanted with osmotic minipumps diffusing either NaCl (NaCl-LUC) or isoproterenol+phenylephrine at 30 mg/kg per day each (isoproterenol+phenylephrine-LUC and isoproterenol + phenylephrine-PDE2A). Minipumps were removed 2 weeks later and mice were euthanized. Cardiac function was assessed throughout the protocol using echocardiography before the injection of the virus and at 2, 3, 4, and 6 weeks. B, Representative blots of PDE2A and GAPDH on the left panel are shown and the bar graph represents ratios of PDE2A over GAPDH quantifications expressed as mean±SEM in a subset of the animals treated with NaCl-LUC, isoproterenol+phenylephrine-LUC, and isoproterenol+phenylephrine-PDE2A. C, Heart weight and lung weight over tibia length ratios expressed as mean±SEM. D, Time course of the normalized ratios of calculated left ventricular weight (over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol+phenylephrine-LUC, and isoproterenol+phenylephrine-PDE2A mice). Number of mice is indicated in the brackets. E, Top panel shows representative images of Masson’s trichrome staining (scale bar 200 μm), the middle panel depicts representative images of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay (scale bar 100 μm) detecting apoptotic nuclei in green costained with the glycocalix marker wheat germ agglutinin in red and nuclei in blue colored with Hoechst. Bar graphs representing mean±SEM quantifications of interstitial fibrosis and apoptotic nuclei in NaCl-LUC (10), isoproterenol+phenylephrine- LUC (9) and isoproterenol+phenylephrine-PDE2A (9) mice are depicted. Statistical significance indicated by the P value (* vs NaCl- LUC and † vs isoproterenol+phenylephrine-LUC) was determined using Welch’s t test (B), Kruskal–Wallis followed by a Dunn’s test (C), 2-way ANOVA followed by a Tukey’s test (D), Welch’s 1-way ANOVA followed by a Dunnett’s test (E). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-Phe-LUC, isoproterenol-phenylephrine-luciferase; Iso-Phe- PDE2A, isoproterenol-phenylephrine-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; ns, not significant; PDE2A, phosphodiesterase 2A; and TL, tibia length.

    Journal: Journal of the American Heart Association

    Article Title: Cardiac Gene Therapy With Phosphodiesterase 2A Limits Remodeling and Arrhythmias in Mouse Models of Heart Failure

    doi: 10.1161/jaha.124.037343

    Figure Lengend Snippet: Figure 3. PDE2A overexpression limits maladaptive remodeling and cardiac dysfunction induced by chronic isoproterenol and phenylephrine infusion. A, Schematic representation of the experimental protocol. Mice were injected with 1012 viral particles of serotype 9 adeno-associated viruses encoding for Luciferase or phosphodiesterase 2A3. Fourteen days later, mice were implanted with osmotic minipumps diffusing either NaCl (NaCl-LUC) or isoproterenol+phenylephrine at 30 mg/kg per day each (isoproterenol+phenylephrine-LUC and isoproterenol + phenylephrine-PDE2A). Minipumps were removed 2 weeks later and mice were euthanized. Cardiac function was assessed throughout the protocol using echocardiography before the injection of the virus and at 2, 3, 4, and 6 weeks. B, Representative blots of PDE2A and GAPDH on the left panel are shown and the bar graph represents ratios of PDE2A over GAPDH quantifications expressed as mean±SEM in a subset of the animals treated with NaCl-LUC, isoproterenol+phenylephrine-LUC, and isoproterenol+phenylephrine-PDE2A. C, Heart weight and lung weight over tibia length ratios expressed as mean±SEM. D, Time course of the normalized ratios of calculated left ventricular weight (over body weight and normalized ejection fraction in NaCl-LUC, isoproterenol+phenylephrine-LUC, and isoproterenol+phenylephrine-PDE2A mice). Number of mice is indicated in the brackets. E, Top panel shows representative images of Masson’s trichrome staining (scale bar 200 μm), the middle panel depicts representative images of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay (scale bar 100 μm) detecting apoptotic nuclei in green costained with the glycocalix marker wheat germ agglutinin in red and nuclei in blue colored with Hoechst. Bar graphs representing mean±SEM quantifications of interstitial fibrosis and apoptotic nuclei in NaCl-LUC (10), isoproterenol+phenylephrine- LUC (9) and isoproterenol+phenylephrine-PDE2A (9) mice are depicted. Statistical significance indicated by the P value (* vs NaCl- LUC and † vs isoproterenol+phenylephrine-LUC) was determined using Welch’s t test (B), Kruskal–Wallis followed by a Dunn’s test (C), 2-way ANOVA followed by a Tukey’s test (D), Welch’s 1-way ANOVA followed by a Dunnett’s test (E). AAV9 indicates serotype 9 adeno-associated viruses; BW, body weight; HW, heart weight; Iso-Phe-LUC, isoproterenol-phenylephrine-luciferase; Iso-Phe- PDE2A, isoproterenol-phenylephrine-phosphodiesterase 2A; LUC, luciferase; LVW, left ventricular weight; LW, lung weight; ns, not significant; PDE2A, phosphodiesterase 2A; and TL, tibia length.

    Article Snippet: The primary antibodies used were as follows: (1) Figure 1: rabbit anti- PDE2A, Fabgennix; PDE2A- 101AP diluted at 1/1000; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; (2) Figure 2: goat antiPDE2A, Santa Cruz, SC- 17228 diluted at 1/1000; mouse anti- vinculin, V9131 Sigma- Aldrich diluted at 1/1000; (3) Figure 3: rabbit anti- PDE2A, Fabgennix, PDE2A- 101AP diluted at 1/1000; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; (4) Figure S1: rabbit anti- flag L5, Novus Biologicals, NBP1- 06712 diluted at 1/500; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; rabbit anti- PDE2A, Proteintech, 55 306- 1- AP diluted at 1/1000; rabbit anti- flag L5, Novus Biologicals, NBP1- 06712 diluted at 1/500; rabbit anti- GAPDH, Cell Signaling, 2118 diluted at 1/1000; (5) Figure S2B: rabbit anti- PDE2A, Proteintech, 55 306- 1- AP diluted at 1/1000; (6) Figure S3: rabbit antiPDE2A, Proteintech, 55 306- 1- AP diluted at 1/1000; and (7) Figure S4: rabbit anti- PDE4A AC55 (gift from Dr. Marco Conti, University of California San Francisco, San Francisco, CA) diluted at 1/1000; rabbit anti- PDE4B (gift from Dr. Marco Conti) diluted at 1/10000; mouse antiPDE4D (gift from Dr Marco Conti) diluted at 1/1000 and rabbit anti- PDE3A (gift from Dr Chen Yan, University of Rochester, Rochester, NY) diluted at 1/10000.

    Techniques: Over Expression, Injection, Luciferase, Virus, Staining, TUNEL Assay, Marker