Src family kinase involvement in rat preglomerular microvascular contractile and [Ca2+]i responses to ANG II

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Experiments were performed to investigate the potential role of Src family kinase(s) in the rat afferent arteriolar contractile response to ANG II. The in vitro blood-perfused juxtamedullary nephron technique was employed to monitor afferent arteriolar lumen diameter responses to 1-100 nM ANG II before and during Src family kinase inhibition (10 μM PP2). PP2 did not alter baseline diameter but attenuated ANG II-induced contractile responses by 33 ± 6%. An inactive analog of PP2 (PP3) had no effect on ANG II-induced afferent arteriolar contraction. The effect of Src kinase inhibition on ANG II-induced intracellular free Ca2+ concentration ([Ca2+]i) responses was probed in fura 2-loaded preglomerular microvascular smooth muscle cells (PVSMCs) obtained from explants and studied after 3-5 days in culture. In untreated PVSMCs, ANG II evoked peak (Δ = 293 ± 66 nM) and plateau (Δ = 23 ± 8 nM) increases in [Ca2+]i. In PVSMCs pretreated with PP2, baseline [Ca2+]i was unaltered, but both the peak (Δ = 140 ± 22 nM) and plateau (Δ = 3 ± 2 nM) phases of the ANG II response were significantly reduced compared with untreated cells. PP3 did not alter [Ca2+]i responses to ANG II. Immunoprecipitation and Western blot analysis confirmed that 100 nM ANG II increased phosphorylation of c-Src (at Y416) in PVSMCs. The phosphorylation response was maximal 1 min after ANG II exposure and was prevented by PP2. We conclude that the preglomerular vasoconstriction evoked by ANG II involves rapid c-Src activation with subsequent effects that contribute to the [Ca2+]i response to the peptide.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Renal Physiology
Volume288
Issue number4 57-4
DOIs
StatePublished - Apr 1 2005

Fingerprint

src-Family Kinases
Smooth Muscle Myocytes
Phosphorylation
Fura-2
Nephrons
Vasoconstriction
Immunoprecipitation
Western Blotting
Peptides

Keywords

  • Afferent arteriole
  • Tyrosine kinase
  • Vascular smooth muscle

ASJC Scopus subject areas

  • Physiology
  • Urology

Cite this

@article{26c39b37bbb44795a29cf1bcf027c15d,
title = "Src family kinase involvement in rat preglomerular microvascular contractile and [Ca2+]i responses to ANG II",
abstract = "Experiments were performed to investigate the potential role of Src family kinase(s) in the rat afferent arteriolar contractile response to ANG II. The in vitro blood-perfused juxtamedullary nephron technique was employed to monitor afferent arteriolar lumen diameter responses to 1-100 nM ANG II before and during Src family kinase inhibition (10 μM PP2). PP2 did not alter baseline diameter but attenuated ANG II-induced contractile responses by 33 ± 6{\%}. An inactive analog of PP2 (PP3) had no effect on ANG II-induced afferent arteriolar contraction. The effect of Src kinase inhibition on ANG II-induced intracellular free Ca2+ concentration ([Ca2+]i) responses was probed in fura 2-loaded preglomerular microvascular smooth muscle cells (PVSMCs) obtained from explants and studied after 3-5 days in culture. In untreated PVSMCs, ANG II evoked peak (Δ = 293 ± 66 nM) and plateau (Δ = 23 ± 8 nM) increases in [Ca2+]i. In PVSMCs pretreated with PP2, baseline [Ca2+]i was unaltered, but both the peak (Δ = 140 ± 22 nM) and plateau (Δ = 3 ± 2 nM) phases of the ANG II response were significantly reduced compared with untreated cells. PP3 did not alter [Ca2+]i responses to ANG II. Immunoprecipitation and Western blot analysis confirmed that 100 nM ANG II increased phosphorylation of c-Src (at Y416) in PVSMCs. The phosphorylation response was maximal 1 min after ANG II exposure and was prevented by PP2. We conclude that the preglomerular vasoconstriction evoked by ANG II involves rapid c-Src activation with subsequent effects that contribute to the [Ca2+]i response to the peptide.",
keywords = "Afferent arteriole, Tyrosine kinase, Vascular smooth muscle",
author = "Qi Che and Carmines, {Pamela K}",
year = "2005",
month = "4",
day = "1",
doi = "10.1152/ajprenal.00392.2004",
language = "English (US)",
volume = "288",
journal = "American Journal of Physiology - Renal Physiology",
issn = "0363-6127",
publisher = "American Physiological Society",
number = "4 57-4",

}

TY - JOUR

T1 - Src family kinase involvement in rat preglomerular microvascular contractile and [Ca2+]i responses to ANG II

AU - Che, Qi

AU - Carmines, Pamela K

PY - 2005/4/1

Y1 - 2005/4/1

N2 - Experiments were performed to investigate the potential role of Src family kinase(s) in the rat afferent arteriolar contractile response to ANG II. The in vitro blood-perfused juxtamedullary nephron technique was employed to monitor afferent arteriolar lumen diameter responses to 1-100 nM ANG II before and during Src family kinase inhibition (10 μM PP2). PP2 did not alter baseline diameter but attenuated ANG II-induced contractile responses by 33 ± 6%. An inactive analog of PP2 (PP3) had no effect on ANG II-induced afferent arteriolar contraction. The effect of Src kinase inhibition on ANG II-induced intracellular free Ca2+ concentration ([Ca2+]i) responses was probed in fura 2-loaded preglomerular microvascular smooth muscle cells (PVSMCs) obtained from explants and studied after 3-5 days in culture. In untreated PVSMCs, ANG II evoked peak (Δ = 293 ± 66 nM) and plateau (Δ = 23 ± 8 nM) increases in [Ca2+]i. In PVSMCs pretreated with PP2, baseline [Ca2+]i was unaltered, but both the peak (Δ = 140 ± 22 nM) and plateau (Δ = 3 ± 2 nM) phases of the ANG II response were significantly reduced compared with untreated cells. PP3 did not alter [Ca2+]i responses to ANG II. Immunoprecipitation and Western blot analysis confirmed that 100 nM ANG II increased phosphorylation of c-Src (at Y416) in PVSMCs. The phosphorylation response was maximal 1 min after ANG II exposure and was prevented by PP2. We conclude that the preglomerular vasoconstriction evoked by ANG II involves rapid c-Src activation with subsequent effects that contribute to the [Ca2+]i response to the peptide.

AB - Experiments were performed to investigate the potential role of Src family kinase(s) in the rat afferent arteriolar contractile response to ANG II. The in vitro blood-perfused juxtamedullary nephron technique was employed to monitor afferent arteriolar lumen diameter responses to 1-100 nM ANG II before and during Src family kinase inhibition (10 μM PP2). PP2 did not alter baseline diameter but attenuated ANG II-induced contractile responses by 33 ± 6%. An inactive analog of PP2 (PP3) had no effect on ANG II-induced afferent arteriolar contraction. The effect of Src kinase inhibition on ANG II-induced intracellular free Ca2+ concentration ([Ca2+]i) responses was probed in fura 2-loaded preglomerular microvascular smooth muscle cells (PVSMCs) obtained from explants and studied after 3-5 days in culture. In untreated PVSMCs, ANG II evoked peak (Δ = 293 ± 66 nM) and plateau (Δ = 23 ± 8 nM) increases in [Ca2+]i. In PVSMCs pretreated with PP2, baseline [Ca2+]i was unaltered, but both the peak (Δ = 140 ± 22 nM) and plateau (Δ = 3 ± 2 nM) phases of the ANG II response were significantly reduced compared with untreated cells. PP3 did not alter [Ca2+]i responses to ANG II. Immunoprecipitation and Western blot analysis confirmed that 100 nM ANG II increased phosphorylation of c-Src (at Y416) in PVSMCs. The phosphorylation response was maximal 1 min after ANG II exposure and was prevented by PP2. We conclude that the preglomerular vasoconstriction evoked by ANG II involves rapid c-Src activation with subsequent effects that contribute to the [Ca2+]i response to the peptide.

KW - Afferent arteriole

KW - Tyrosine kinase

KW - Vascular smooth muscle

UR - http://www.scopus.com/inward/record.url?scp=15044347565&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=15044347565&partnerID=8YFLogxK

U2 - 10.1152/ajprenal.00392.2004

DO - 10.1152/ajprenal.00392.2004

M3 - Article

VL - 288

JO - American Journal of Physiology - Renal Physiology

JF - American Journal of Physiology - Renal Physiology

SN - 0363-6127

IS - 4 57-4

ER -