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Am J Physiol Renal Physiol (November 30, 2004). doi:10.1152/ajprenal.00392.2004
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Submitted on October 26, 2004
Accepted on November 26, 2004

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

Qi Che1 and Pamela K. Carmines1*

1 Department of Cellular & Integrative Physiology, University of Nebraska College of Medicine, Omaha, NE, USA

* To whom correspondence should be addressed. E-mail: pcarmines{at}unmc.edu.

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 [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 ({Delta} = 295 ± 61 nM) and plateau ({Delta} = 29 ± 9 nM) increases in [Ca2+]i. In PVSMCs pre-treated with PP2, baseline [Ca2+]i was unaltered but both the peak ({Delta} = 140 ± 22 nM) and plateau ({Delta} = 5 ± 3 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.




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