http://hdl.handle.net/10027/1242 Thu, 23 May 2013 05:23:39 GMT 2013-05-23T05:23:39Z http://hdl.handle.net/10027/8727 Characterization of dual agonists for kinin B1 and B2 receptors and their biased activation of B2 receptors Brovkoyvch, Viktor; Lowry, Jessica L.; Skidgel, Randal A.; Zhang, Xianming Kinin B1 and B2 receptors (kB1R and kB2R) play important roles in many physiological and pathological processes. In some cases, kB1R or kB2R activation can have overlapping or complementary beneficial effects, thus an activator of both receptors might be advantageous. We found that replacement of the C-terminal Arg in the natural kB2R activators bradykinin (BK) or kallidin (KD) with Lys (K9-BK or K10-KD) resulted in agonists that effectively stimulate the downstream signaling of both the kB1R and kB2R as measured by increased inositol turnover, intracellular calcium, ERK1/2 phosphorylation, arachidonic acid release and NO production. However, K9-BK and K10-KD displayed some characteristics of biased agonism for kB2Rs as indicated by the rapid kinetics of ERK1/2 phosphorylation induced by K9-BK or K10-KD compared with the prolonged response mediated by BK or KD. In contrast, kinetics of ERK phosphorylation stimulated by K10-KD activation of the kB1R was as same as that induced by known kB1R agonist des-Arg10-KD. Furthermore, the endocytosis of kB2Rs mediated by K9- BK and K10-KD was remarkably less than that induced by BK and KD respectively. K10-KD stimulated kB1R and kB2R-dependent calcium responses and ERK1/2 phosphorylation in bovine endothelial cells. In cytokine-treated human endothelial cells, K10-KD stimulated ERK1/2 phosphorylation and a transient peak of NO production that was primarily kB2R-dependent. K10-KD also stimulated prolonged NO production that was both kB1R and kB2R-dependent. These data provide the first examples of dual agonists of kB1R and kB2R, and a biased agonist of kB2R and may provide useful clues for developing dual modulators of kB1Rs and kB2Rs for potential therapeutic use. NOTICE: this is the author’s version of a work that was accepted for publication in Cellular Signalling. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in PUBLICATION, [Vol 24, Issue 8, 2012 Aug] DOI: 10.1016/j.cellsig.2012.04.002 Wed, 01 Aug 2012 05:00:00 GMT http://hdl.handle.net/10027/8727 2012-08-01T05:00:00Z http://hdl.handle.net/10027/8680 Intersectin 1 contributes to phenotypes in vivo: implications for Down Syndrome Hunter, Michael P.; Nelson, Marianela; Kurzer, Michael; Wang, Xuerong; Kryscio, Richard J.; Head, Elizabeth; Pinna, Graziano; O’Bryan, John P. Intersectin 1 (ITSN1) is a chromosome 21 (HSA21) gene product encoding a multidomain scaffold protein that functions in endocytosis, signal transduction and is implicated in Down Syndrome, Alzheimer’s Disease, and potentially other neurodegenerative diseases through activation of c-Jun N-terminal kinase (JNK). We report for the first time that ITSN1 proteins are elevated in Down Syndrome individuals of varying ages. However, ITSN1 levels decreased in aged Down Syndrome cases with Alzheimer’s Disease-like neuropathology. Analysis of a novel ITSN1 transgenic mouse reveals that ITSN1 overexpression results in a sex-dependent decrease in locomotor activity. This study reveals a link between overexpression of specific ITSN1 isoforms and behavioral phenotypes and has implications for human neurodegenerative diseases such as Down Syndrome and Alzheimer’s Disease. This is a copy of an article published in the NeuroReport © 2011 Lippincott, Williams & Wilkins. Post print version of article may differ from published version. The final publication is available at www.lww.com/; DOI: 10.1097/WNR.0b013e32834ae348 Sat, 01 Oct 2011 05:00:00 GMT http://hdl.handle.net/10027/8680 2011-10-01T05:00:00Z http://hdl.handle.net/10027/8580 Activity of Ca -activated Cl channels contributes to regulating receptor- and store-operated Ca entry in human pulmonary artery smooth muscle cells Yamamura, Aya; Yamamura, Hisao; Zeifman, Amy; Yuan, Jason X-J Intracellular Ca(2+) plays a fundamental role in regulating cell functions in pulmonary arterial smooth muscle cells (PASMCs). A rise in cytosolic Ca(2+) concentration ([Ca(2+)](cyt)) triggers pulmonary vasoconstriction and stimulates PASMC proliferation. [Ca(2+)](cyt) is increased mainly by Ca(2+) release from intracellular stores and Ca(2+) influx through plasmalemmal Ca(2+)-permeable channels. Given the high concentration of intracellular Cl(-) in PASMCs, Ca(2+)-activated Cl(-)(Cl(Ca)) channels play an important role in regulating membrane potential and cell excitability of PASMCs. In this study, we examined whether activity of Cl(Ca) channels was involved in regulating [Ca(2+)](cyt) in human PASMCs via regulating receptor- (ROCE) and store- (SOCE) operated Ca(2+) entry. The data demonstrated that an angiotensin II (100 nM)-mediated increase in [Ca(2+)](cyt) via ROCE was markedly attenuated by the Cl(Ca) channel inhibitors, niflumic acid (100 muM), flufenamic acid (100 muM), and 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (100 muM). The inhibition of Cl(Ca) channels by niflumic acid and flufenamic acid significantly reduced both transient and plateau phases of SOCE that was induced by passive depletion of Ca(2+) from the sarcoplasmic reticulum by 10 muM cyclopiazonic acid. In addition, ROCE and SOCE were abolished by SKF-96365 (50 muM) and 2-aminoethyl diphenylborinate (100 muM), and were slightly decreased in the presence of diltiazem (10 muM). The electrophysiological and immunocytochemical data indicate that Cl(Ca) currents were present and TMEM16A was functionally expressed in human PASMCs. The results from this study suggest that the function of Cl(Ca) channels, potentially formed by TMEM16A proteins, contributes to regulating [Ca(2+)](cyt) by affecting ROCE and SOCE in human PASMCs. The original version is available through Medknow Publications at DOI: 10.4103/2045-8932.83447 Fri, 01 Apr 2011 05:00:00 GMT http://hdl.handle.net/10027/8580 2011-04-01T05:00:00Z http://hdl.handle.net/10027/8573 Improvement of cardiac function in mouse myocardial infarction after transplantation of epigenetically-modified bone marrow progenitor cells Rajasingh, Johnson; Thangavel, Jayakumar; Siddiqui, Mohammad R.; Gomes, Ignatius; Gao, Xiao-pei; Kishore, Raj; Malik, Asrar B. Objective To study usefulness of bone marrow progenitor cells (BPCs) epigenetically altered by chromatin modifying agents in mediating heart repair after myocardial infarction in mice. Methods and Results We tested the therapeutic efficacy of bone marrow progenitor cells treated with the clinically-used chromatin modifying agents Trichostatin A (TSA, histone deacetylase inhibitor) and 5Aza-2-deoxycytidine (Aza, DNA methylation inhibitor) in a mouse model of acute myocardial infarction (AMI). Treatment of BPCs with Aza and TSA induced expression of pluripotent genes Oct4, Nanog, Sox2, and thereafter culturing these cells in defined cardiac myocyte-conditioned medium resulted in their differentiation into cardiomyocyte progenitors and subsequently into cardiac myocytes. Their transition was deduced by expression of repertoire of markers: Nkx2.5, GATA4, cardiotroponin T, cardiotroponin I, α-sarcomeric actinin, Mef2c and MHC-α. We observed that the modified BPCs had greater AceH3K9 expression and reduced histone deacetylase1 (HDAC1) and lysine-specific demethylase1 (LSD1) expression compared to untreated BPCs, characteristic of epigenetic changes. Intra-myocardial injection of modified BPCs after AMI in mice significantly improved left ventricular function. These changes were ascribed to differentiation of the injected cells into cardiomyocytes and endothelial cells. Conclusion Treatment of BPCs with Aza and TSA converts BPCs into multipotent cells, which can then be differentiated into myocyte progenitors. Transplantation of these modified progenitor cells into infarcted mouse hearts improved left ventricular function secondary to differentiation of cells in the niche into myocytes and endothelial cells. © 2011 Rajasingh et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. DOI: 10.1371/journal.pone.0022550. Fri, 22 Jul 2011 05:00:00 GMT http://hdl.handle.net/10027/8573 2011-07-22T05:00:00Z