Oleg Palygin1, Bradley Miller2, Daria V Ilatovskaya1, Andrey Sorokin3, Alexander Staruschenko4. 1. Department of Physiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA. 2. Department of Medicine, Medical College of Wisconsin, Milwaukee, WI 53226, USA. 3. Department of Medicine, Medical College of Wisconsin, Milwaukee, WI 53226, USA; Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI 53226, USA. Electronic address: Sorokin@mcw.edu. 4. Department of Physiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA; Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
Abstract
AIMS: Endothelin-1 (ET-1) is a potent vasoconstrictor which regulates the physiology of cardiorenal system. The aim of this study was to evaluate ET-1-mediated elevation of intracellular Ca(2+) in smooth muscle cells (SMC) of renal resistance arteries. MAIN METHODS: In in vitro studies of primary SMC, which were isolated from rat renal microvessels, the levels of intracellular Ca(2+) were calculated from the ratio of emissions at 340 and 380nm after loading cells with Fura 2-AM dye. In ex vivo studies we used two-photon imaging of renal resistance arteries excised from rat kidneys and loaded with fluorescent Ca(2+) indicator Fluo-4 AM. KEY FINDINGS: The two-photon imaging demonstrates that treatment of isolated rat renal resistance arteries with ET-1 causes a rapid increase of intracellular Ca(2+) concentration in smooth muscle vasculature of these vessels. These ex vivo observations are in accordance with in vitro findings indicating that ET-1 mediates activation of TRPC channels and increases the level of intracellular Ca(2+) in cultured SMC to 510±83nM. SIGNIFICANCE: ET-1-mediated elevation of intracellular Ca(2+) is strongly linked to renal microvascular contraction and is crucial for ET-1-induced contraction of SMC. The two-photon imaging of intracellular Ca(2+) in intact SMC of rat renal resistance arteries is a powerful technique which allows the detailed ex vivo analysis of intracellular Ca(2+) handling by ET-1, an important player in hypertension-related kidney diseases.
AIMS: Endothelin-1 (ET-1) is a potent vasoconstrictor which regulates the physiology of cardiorenal system. The aim of this study was to evaluate ET-1-mediated elevation of intracellular Ca(2+) in smooth muscle cells (SMC) of renal resistance arteries. MAIN METHODS: In in vitro studies of primary SMC, which were isolated from rat renal microvessels, the levels of intracellular Ca(2+) were calculated from the ratio of emissions at 340 and 380nm after loading cells with Fura 2-AM dye. In ex vivo studies we used two-photon imaging of renal resistance arteries excised from rat kidneys and loaded with fluorescent Ca(2+) indicator Fluo-4 AM. KEY FINDINGS: The two-photon imaging demonstrates that treatment of isolated rat renal resistance arteries with ET-1 causes a rapid increase of intracellular Ca(2+) concentration in smooth muscle vasculature of these vessels. These ex vivo observations are in accordance with in vitro findings indicating that ET-1 mediates activation of TRPC channels and increases the level of intracellular Ca(2+) in cultured SMC to 510±83nM. SIGNIFICANCE: ET-1-mediated elevation of intracellular Ca(2+) is strongly linked to renal microvascular contraction and is crucial for ET-1-induced contraction of SMC. The two-photon imaging of intracellular Ca(2+) in intact SMC of rat renal resistance arteries is a powerful technique which allows the detailed ex vivo analysis of intracellular Ca(2+) handling by ET-1, an important player in hypertension-related kidney diseases.
Authors: Joshua S Speed; J Brett Heimlich; Kelly A Hyndman; Brandon M Fox; Vivek Patel; Masashi Yanagisawa; Jennifer S Pollock; Jens M Titze; David M Pollock Journal: FASEB J Date: 2015-08-12 Impact factor: 5.191
Authors: Daria V Ilatovskaya; Oleg Palygin; Vladislav Chubinskiy-Nadezhdin; Yuri A Negulyaev; Rong Ma; Lutz Birnbaumer; Alexander Staruschenko Journal: Kidney Int Date: 2014-03-19 Impact factor: 10.612
Authors: Bradley Miller; Oleg Palygin; Victoriya A Rufanova; Andrew Chong; Jozef Lazar; Howard J Jacob; David Mattson; Richard J Roman; Jan M Williams; Allen W Cowley; Aron M Geurts; Alexander Staruschenko; John D Imig; Andrey Sorokin Journal: J Clin Invest Date: 2016-06-06 Impact factor: 14.808
Authors: John F LaDisa; Aoy Tomita-Mitchell; Karl Stamm; Kathleen Bazan; Donna K Mahnke; Mary A Goetsch; Brandon J Wegter; Jesse W Gerringer; Kathryn Repp; Oleg Palygin; Adrian P Zietara; Mary M Krolikowski; Thomas J Eddinger; Abdel A Alli; Michael E Mitchell Journal: Physiol Genomics Date: 2019-04-19 Impact factor: 4.297