Studies on the role of connexin 43 phosphorylation in the injury - resistant heart
| dc.contributor.author | Srisakuldee, Wattamon | |
| dc.contributor.examiningcommittee | Zahradka, Peter (Physiology) Netticadan, Thomas (Physiology) Hatch, Grant (Pharmacology & Therapeutics) Tuana, Balwant (University of Ottawa) | en_US |
| dc.contributor.supervisor | Kardami, Elissavet (Human Anatomy & Cell Science) | en_US |
| dc.date.accessioned | 2014-08-19T15:45:57Z | |
| dc.date.available | 2014-08-19T15:45:57Z | |
| dc.date.issued | 2009-09-01 | en_US |
| dc.date.issued | 2014-07-01 | en_US |
| dc.degree.discipline | Physiology | en_US |
| dc.degree.level | Doctor of Philosophy (Ph.D.) | en_US |
| dc.description.abstract | Ischemic heart disease is a major cause of death worldwide. Identifying the mechanisms mediating cardiac resistance to injury (‘cardioprotection’) can contribute to therapies for cardiac injury. These studies investigated the role of the membrane channel forming protein connexin43 (Cx43), a downstream effector of PKCε-mediated cardioprotection. In isolated cardiomyocytes FGF-2/PKCε –induced cytoprotection is mediated by Cx43 phosphorylation at PKCε-target site, serine(S)262. Hypothesis (1). PKCε-mediated cardioprotection increases Cx43 phosphorylation at PKCε-target sites and prevents ischemia and/or reperfusion-induced Cx43 remodeling. Rat hearts were subjected to protective treatments (ischemic preconditioning, FGF-2, diazoxide), followed by 30 min global ischemia and/or 60 min reperfusion. All treatments elicited above-physiological levels of phospho(p)S262-Cx43 and pS368-Cx43 (P*Cx43), which were sustained during global ischemia, and reperfusion, and accompanied by attenuation of ischemia-induced Cx43 dephosphorylation, and prevention of Cx43 lateralization. Cx43 is present in cardiac subsarcolemmal (SSM) but not interfibrillar (IFM) mitochondria. Hypothesis (2). FGF-2 exerts protective effects on both mitochondrial populations, but is associated with mitochondrial (mito) P*Cx43 state in SSM, and mediated by mitoCx43 function. FGF-2 treatment increased calcium tolerance in SSM and IFM by 2.9- and 1.7-fold, respectively, compared to controls. In the presence of Gap27, a Cx43 hemi-channel blocker, the salutary effect of FGF-2 were lost in SSM but not IFM, indicating a functional role for Cx43. FGF-2 increased levels of PKCε, pPKCε and Tom-20 translocase in SSM and IFM. In SSM, FGF-2 increased pS262-and pS368-Cx43 by 30-fold and 8-fold, compared to controls. Stimulation of untreated SSM with a PKC activator (phorbol 12-myristate 13-acetate; PMA) also increased pS262-and-S368-Cx43 and calcium tolerance, which was prevented by εV1-2, a PKCε-inhibiting peptide. The effect of FGF-2 on isolated cardiac mitochondria is unknown. Hypothesis (3). FGF-2 exerts a direct protective effects on SSM. Direct stimulation of SSM with FGF-2 increased pS262-and-S368-mitoCx43, and calcium resistance to mPTP, and was dependent on mitoPKCε. FGF receptor inhibitors, SU5402 and FGFR neutralizing antibodies, blocked the effect of FGF-2, suggesting a FGFR1-like protein is responsible for the direct protective effects of FGF-2. This new intracellular mechanism of cytoprotection implies that endogenous intracellular levels of FGF-2 may determine constitutive levels of cardiac mitochondrial resistance to mPTP. | en_US |
| dc.description.note | October 2014 | en_US |
| dc.identifier.citation | Phosphorylation of connexin-43 at serine 262 promotes a cardiac injury-resistant state. Srisakuldee W, Jeyaraman MM, Nickel BE, Tanguy S, Jiang ZS, Kardami E. Cardiovasc Res. 2009 Sep 1;83(4):672-81. | en_US |
| dc.identifier.citation | The FGF-2-triggered protection of cardiac subsarcolemmal mitochondria from calcium overload is mitochondrial connexin 43-dependent. Srisakuldee W, Makazan Z, Nickel BE, Zhang F, Thliveris JA, Pasumarthi KB, Kardami E. Cardiovasc Res. 2014 Jul 1;103(1):72-80. | en_US |
| dc.identifier.uri | http://hdl.handle.net/1993/23823 | |
| dc.language.iso | eng | en_US |
| dc.publisher | Oxford journals (Cardiovascular Research) | en_US |
| dc.publisher | Oxford journals (Cardiovascular Research) | en_US |
| dc.rights | open access | en_US |
| dc.subject | connexin 43 | en_US |
| dc.subject | cardioprotection | en_US |
| dc.subject | mitochondria | en_US |
| dc.title | Studies on the role of connexin 43 phosphorylation in the injury - resistant heart | en_US |
| dc.type | doctoral thesis | en_US |