Protein kinase N2 mediates flow-induced endothelial NOS activation and vascular tone regulation

Formation of NO by endothelial NOS (eNOS) is a central process in the homeostatic regulation of vascular functions including blood pressure regulation, and fluid shear stress exerted by the flowing blood is a main stimulus of eNOS activity. Previous work has identified several mechanosensing and -transducing processes in endothelial cells, which mediate this process and induce the stimulation of eNOS activity through phosphorylation of the enzyme via various kinases including AKT. How the initial mechanosensing and signaling processes are linked to eNOS phosphorylation is unclear. In human endothelial cells, we demonstrated that protein kinase N2 (PKN2), which is activated by flow through the mechanosensitive cation channel Piezo1 and G_q/G₁₁-mediated signaling, as well as by Ca²⁺ and phosphoinositide-dependent protein kinase 1 (PDK1), plays a pivotal role in this process. Active PKN2 promoted the phosphorylation of human eNOS at serine 1177 and at a newly identified site, serine 1179. These phosphorylation events additively led to increased eNOS activity. PKN2-mediated eNOS phosphorylation at serine 1177 involved the phosphorylation of AKT synergistically with mTORC2-mediated AKT phosphorylation, whereas active PKN2 directly phosphorylated human eNOS at serine 1179. Mice with induced endothelium-specific deficiency of PKN2 showed strongly reduced flow-induced vasodilation and developed arterial hypertension accompanied by reduced eNOS activation. These results uncover a central mechanism that couples upstream mechanosignaling processes in endothelial cells to the regulation of eNOS-mediated NO formation, vascular tone, and blood pressure.     

A new pulse sequence for determining T1 and T2 simultaneously

Determination of the relaxation times T1 and T2 which are important for tissue characterization generally requires the use of different pulse sequences in magnetic resonance imaging. In this study, a new pulse sequence which facilitates simultaneous determination of the T1 and T2 times is presented. Determination takes place in this case pixel by pixel from the measured images. The measuring time corresponds in this case approximately to that of a normal spin-echo sequence with long repetition time and two data acquisitions. The functional dependence of the accuracy of the T1 and T2 determination upon external errors, e.g., angle of rotation errors, is discussed. The tissue contrast behavior of the individual echoes is shown and its dependence on pulse parameters is explained.     

Pathologische Anatomie der neuro-endokrinen Erkrankungen

Ohne ZusammenfassungMit 14 TextabbildungenHerrn Prof. Dr. H. J. Deuticke zum 60. Geburtstag.Mit Unterstützung der deutschen Forschungsgemeinschaft.     

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