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Calcium-dependent signalling is essential during collateral growth in the pig hind limb-ischemia model Troidl C; Nef H; Voss S; Schilp A; Kostin S; Troidl K; Szardien S; Rolf A; Schmitz-Rixen T; Schaper W; Hamm CW; Elsasser A; Mollmann HJ Mol Cell Cardiol 2010[Jul]; 49 (1): 142-51We investigated the effect of pharmacological activation of the Ca(2+)-channel transient receptor potential cation channel, subfamily V, member 4 (TRPV4) on collateral growth in a pig hind limb-ischemia model thereby identifying subcellular mechanisms. Domestic pigs received femoral artery ligature and were randomly assigned to one of the following groups (each n=6): (1) 4alpha-phorbol 12,13-didecanoate (4alphaPDD) treatment; (2) treatment with an arterio-venous shunt (AV-shunt) distal to the occlusion; or (3) implantation of NaCl-filled minipump. Six sham-operated pigs acted as controls. Aortic and peripheral mean arterial pressure (MAP) measurements were performed to assess the collateral flow index (CFI). Tissue was isolated from M. quadriceps for immunohistochemistry and from isolated collateral arteries for quantitative real time PCR (qRT-PCR). Shortly after ligature the CFI dropped from 0.96+/-0.02 to 0.21+/-0.02 in all ligature-treated groups. In ligature-only-treated pigs CFI increased to 0.56+/-0.03 after 7days. Treatment with 4alphaPDD led to an enhancement of CFI compared with ligature alone (0.73+/-0.03). CD31-staining showed improved arteriolar density. Increased Ki67 staining in collaterals indicated proliferation. qRT-PCR and Western blot analysis showed upregulation or modulation of Ca(2+)-dependent transcription factors nuclear factor of activated T-cells, cytoplasmic, calcineurin-dependent 1 (NFATc1), Kv channel interacting protein 3, calsenilin (KCNIP3/CSEN/DREAM), and myocyte enhancer factor 2C (MEF2C) in 4alphaPDD- and AV-shunt-treated pigs compared with controls. Improved CFI after 4alphaPDD treatment identifies TRPV4 as an initial fluid shear-stress sensor and collateral remodelling and growth trigger. Subcellularly, modulation of Ca(2+)-dependent transcription factors indicates a pivotal role for Ca(2+)-signalling during arteriogenesis.|Animals[MESH]|Aorta/metabolism/physiopathology[MESH]|Arteries/metabolism/physiopathology[MESH]|Blood Vessels/metabolism/physiopathology[MESH]|Calcium Signaling[MESH]|Femoral Artery/metabolism/physiopathology/surgery[MESH]|Hindlimb/*blood supply/metabolism/physiopathology[MESH]|Ischemia/metabolism/*physiopathology[MESH]|Lower Extremity/blood supply/physiopathology[MESH]|Male[MESH]|NFATC Transcription Factors/metabolism/pharmacology[MESH]|Phorbols[MESH]|Random Allocation[MESH]|Stress, Mechanical[MESH]|Sus scrofa/metabolism[MESH] |
