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Defining the roles of arrestin2 and arrestin3 in vasoconstrictor receptor desensitization in hypertension.

Willets JM, Nash CA, Rainbow RD, Nelson CP, Challiss RA - Am. J. Physiol., Cell Physiol. (2015)

Bottom Line: Desensitization of UTP-stimulated vessel contractions was increased in 12-wk-old (but not 6-wk-old) SHR animals.In conclusion, arrestin2 and 3 expression is elevated in resistance arteries during the emergence of the early hypertensive phenotype, which underlies an enhanced ability to desensitize vasoconstrictor signaling and vessel contraction.Such regulatory changes may act to compensate for increased vasoconstrictor-induced vessel contraction.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Physiology and Pharmacology, University of Leicester, Leicester, United Kingdom; and jmw23@leicester.ac.uk.

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Specificity of arrestin isoformic knockdown. Arterial smooth muscle cells were transfected with 10 nM negative control (NC), anti-arrestin2 or anti-arrestin3 short-interfering RNAs (siRNAs) using the nucleofection technique. After 48 h, cells were lysed and 40 μg of each sample were loaded onto 10% SDS-PAGE gels for separation and arrestin expression was determined using immunoblotting as described in materials and methods. A: representative immunoblots showing the effect of individual siRNA treatments on arrestin expression. B and C: cumulative data (means ± SE for n = 5 cell preparations from 5 separate animals from each strain) showing the degree of arrestin2 (B) and arrestin3 (C) knockdown following siRNA treatments in comparison to GAPDH expression. Statistical significance is indicated as ***P < 0.001 for arrestin2 knockdown and **P < 0.01 for arrestin3 knockdown compared with negative control transfected cells (two-way ANOVA and Tukey's post hoc test).
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Figure 7: Specificity of arrestin isoformic knockdown. Arterial smooth muscle cells were transfected with 10 nM negative control (NC), anti-arrestin2 or anti-arrestin3 short-interfering RNAs (siRNAs) using the nucleofection technique. After 48 h, cells were lysed and 40 μg of each sample were loaded onto 10% SDS-PAGE gels for separation and arrestin expression was determined using immunoblotting as described in materials and methods. A: representative immunoblots showing the effect of individual siRNA treatments on arrestin expression. B and C: cumulative data (means ± SE for n = 5 cell preparations from 5 separate animals from each strain) showing the degree of arrestin2 (B) and arrestin3 (C) knockdown following siRNA treatments in comparison to GAPDH expression. Statistical significance is indicated as ***P < 0.001 for arrestin2 knockdown and **P < 0.01 for arrestin3 knockdown compared with negative control transfected cells (two-way ANOVA and Tukey's post hoc test).

Mentions: Western blotting analysis indicated that arrestin2 or arrestin3 siRNA treatments induced isoform-specific knockdowns of ≥85% and ≥80%, respectively, in both WKY- and SHR-derived cells (Fig. 7). Following MSMC transfection with negative-control siRNA, R2/R1 ratios in both WKY- and SHR-derived cells (Fig. 8, A, B, and E) were similar to those obtained in MSMC not transfected with siRNAs (see Fig. 6).


Defining the roles of arrestin2 and arrestin3 in vasoconstrictor receptor desensitization in hypertension.

Willets JM, Nash CA, Rainbow RD, Nelson CP, Challiss RA - Am. J. Physiol., Cell Physiol. (2015)

Specificity of arrestin isoformic knockdown. Arterial smooth muscle cells were transfected with 10 nM negative control (NC), anti-arrestin2 or anti-arrestin3 short-interfering RNAs (siRNAs) using the nucleofection technique. After 48 h, cells were lysed and 40 μg of each sample were loaded onto 10% SDS-PAGE gels for separation and arrestin expression was determined using immunoblotting as described in materials and methods. A: representative immunoblots showing the effect of individual siRNA treatments on arrestin expression. B and C: cumulative data (means ± SE for n = 5 cell preparations from 5 separate animals from each strain) showing the degree of arrestin2 (B) and arrestin3 (C) knockdown following siRNA treatments in comparison to GAPDH expression. Statistical significance is indicated as ***P < 0.001 for arrestin2 knockdown and **P < 0.01 for arrestin3 knockdown compared with negative control transfected cells (two-way ANOVA and Tukey's post hoc test).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4525080&req=5

Figure 7: Specificity of arrestin isoformic knockdown. Arterial smooth muscle cells were transfected with 10 nM negative control (NC), anti-arrestin2 or anti-arrestin3 short-interfering RNAs (siRNAs) using the nucleofection technique. After 48 h, cells were lysed and 40 μg of each sample were loaded onto 10% SDS-PAGE gels for separation and arrestin expression was determined using immunoblotting as described in materials and methods. A: representative immunoblots showing the effect of individual siRNA treatments on arrestin expression. B and C: cumulative data (means ± SE for n = 5 cell preparations from 5 separate animals from each strain) showing the degree of arrestin2 (B) and arrestin3 (C) knockdown following siRNA treatments in comparison to GAPDH expression. Statistical significance is indicated as ***P < 0.001 for arrestin2 knockdown and **P < 0.01 for arrestin3 knockdown compared with negative control transfected cells (two-way ANOVA and Tukey's post hoc test).
Mentions: Western blotting analysis indicated that arrestin2 or arrestin3 siRNA treatments induced isoform-specific knockdowns of ≥85% and ≥80%, respectively, in both WKY- and SHR-derived cells (Fig. 7). Following MSMC transfection with negative-control siRNA, R2/R1 ratios in both WKY- and SHR-derived cells (Fig. 8, A, B, and E) were similar to those obtained in MSMC not transfected with siRNAs (see Fig. 6).

Bottom Line: Desensitization of UTP-stimulated vessel contractions was increased in 12-wk-old (but not 6-wk-old) SHR animals.In conclusion, arrestin2 and 3 expression is elevated in resistance arteries during the emergence of the early hypertensive phenotype, which underlies an enhanced ability to desensitize vasoconstrictor signaling and vessel contraction.Such regulatory changes may act to compensate for increased vasoconstrictor-induced vessel contraction.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Physiology and Pharmacology, University of Leicester, Leicester, United Kingdom; and jmw23@leicester.ac.uk.

Show MeSH
Related in: MedlinePlus