Rania E Mufti1, Anil Zechariah1, Maria Sancho1, Neil Mazumdar1, Suzanne E Brett1, Donald G Welsh2. 1. From the Hotchkiss Brain Institute (R.E.M., A.Z., M.S., N.M., S.E.B., D.G.W.), Libin Cardiovascular Institute (R.E.M., A.Z., M.S., N.M., S.E.B., D.G.W.), and Department of Physiology and Pharmacology (R.E.M., A.Z., M.S., N.M., S.E.B., D.G.W.), University of Calgary, Alberta, Canada; and Department of Physiology and Pharmacology, University of Western Ontario, London, Ontario, Canada (A.Z., M.S., N.M., S.E.B., D.G.W.). 2. From the Hotchkiss Brain Institute (R.E.M., A.Z., M.S., N.M., S.E.B., D.G.W.), Libin Cardiovascular Institute (R.E.M., A.Z., M.S., N.M., S.E.B., D.G.W.), and Department of Physiology and Pharmacology (R.E.M., A.Z., M.S., N.M., S.E.B., D.G.W.), University of Calgary, Alberta, Canada; and Department of Physiology and Pharmacology, University of Western Ontario, London, Ontario, Canada (A.Z., M.S., N.M., S.E.B., D.G.W.). dwelsh@ucalgary.ca.
Abstract
OBJECTIVE: The myogenic response is central to blood flow regulation in the brain. Its induction is tied to elevated cytosolic [Ca(2+)], a response primarily driven by voltage-gated Ca(2+) channels and secondarily by Ca(2+) wave production. Although the signaling events leading to the former are well studied, those driving Ca(2+) waves remain uncertain. APPROACH AND RESULTS: We postulated that αvβ3 integrin signaling is integral to the generation of pressure-induced Ca(2+) waves and cerebral arterial tone. This hypothesis was tested in rat cerebral arteries using the synergistic strengths of pressure myography, rapid Ca(2+) imaging, and Western blot analysis. GRGDSP, a peptide that preferentially blocks αvβ3 integrin, attenuated myogenic tone, indicating the modest role for sarcoplasmic reticulum Ca(2+) release in myogenic tone generation. The RGD peptide was subsequently shown to impair Ca(2+) wave generation and myosin light chain 20 (MLC20) phosphorylation, the latter of which was attributed to the modulation of MLC kinase and MLC phosphatase via MYPT1-T855 phosphorylation. Subsequent experiments revealed that elevated pressure enhanced phospholipase Cγ1 phosphorylation in an RGD-dependent manner and that phospholipase C inhibition attenuated Ca(2+) wave generation. Direct inhibition of inositol 1, 4, 5-triphosphate receptors also impaired Ca(2+) wave generation, myogenic tone, and MLC20 phosphorylation, partly through the T-855 phosphorylation site of MYPT1. CONCLUSIONS: Our investigation reveals a hitherto unknown role for αvβ3 integrin as a cerebral arterial pressure sensor. The membrane receptor facilitates Ca(2+) wave generation through a signaling cascade, involving phospholipase Cγ1, inositol 1,3,4 triphosphate production, and inositol 1, 4, 5-triphosphate receptor activation. These discrete asynchronous Ca(2+) events facilitate MLC20 phosphorylation and, in part, myogenic tone by influencing both MLC kinase and MLC phosphatase activity.
OBJECTIVE: The myogenic response is central to blood flow regulation in the brain. Its induction is tied to elevated cytosolic [Ca(2+)], a response primarily driven by voltage-gated Ca(2+) channels and secondarily by Ca(2+) wave production. Although the signaling events leading to the former are well studied, those driving Ca(2+) waves remain uncertain. APPROACH AND RESULTS: We postulated that αvβ3 integrin signaling is integral to the generation of pressure-induced Ca(2+) waves and cerebral arterial tone. This hypothesis was tested in rat cerebral arteries using the synergistic strengths of pressure myography, rapid Ca(2+) imaging, and Western blot analysis. GRGDSP, a peptide that preferentially blocks αvβ3 integrin, attenuated myogenic tone, indicating the modest role for sarcoplasmic reticulum Ca(2+) release in myogenic tone generation. The RGD peptide was subsequently shown to impair Ca(2+) wave generation and myosin light chain 20 (MLC20) phosphorylation, the latter of which was attributed to the modulation of MLC kinase and MLC phosphatase via MYPT1-T855 phosphorylation. Subsequent experiments revealed that elevated pressure enhanced phospholipase Cγ1 phosphorylation in an RGD-dependent manner and that phospholipase C inhibition attenuated Ca(2+) wave generation. Direct inhibition of inositol 1, 4, 5-triphosphate receptors also impaired Ca(2+) wave generation, myogenic tone, and MLC20 phosphorylation, partly through the T-855 phosphorylation site of MYPT1. CONCLUSIONS: Our investigation reveals a hitherto unknown role for αvβ3 integrin as a cerebral arterial pressure sensor. The membrane receptor facilitates Ca(2+) wave generation through a signaling cascade, involving phospholipase Cγ1, inositol 1,3,4 triphosphate production, and inositol 1, 4, 5-triphosphate receptor activation. These discrete asynchronous Ca(2+) events facilitate MLC20 phosphorylation and, in part, myogenic tone by influencing both MLC kinase and MLC phosphatase activity.
Authors: Anil Zechariah; Cam Ha T Tran; Bjorn O Hald; Shaun L Sandow; Maria Sancho; Michelle Sun Mi Kim; Sergio Fabris; Ursula I Tuor; Grant R J Gordon; Donald G Welsh Journal: Arterioscler Thromb Vasc Biol Date: 2019-12-12 Impact factor: 8.311
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