The role of NO/sGC signaling in erythropoiesis: Lessons from erythroid-specific knock-out and knock-in mice

The nitric oxide (NO)/soluble guanylyl cyclase (sGC)signaling pathway in human erythroid cells has been proposed to drive theexpression of γ-globin and control erythropoiesis, but its role in vivo has not yet been demonstrated.We generated erythroid cell-specific eNOS knock-out and knock-in mice, as well as erythroid-specific sGC KO mice, using the loxP/Cre recombinase gene targeting approach. We confirmed the specificity of gene targetingby analyzing erythroid cell-specific DNA recombination, target protein expression, and/or activity using qPCR, western blotting, ELISA, and enzymatic assays.To analyze changes in the hematopoietic phenotype, we analyzed the peripheral blood cell and reticulocyte counts. In the bone marrow and spleen of the mice, we analyzed cellular composition, quantified stage-specific cell differentiation by flow cytometry, and performed a functional colony assay. Moreover, we determined the vascular function, systemic hemodynamics,andNO metabolites.This talk will present the results of these recent and ongoing investigations analyzing the role of the NO/sGC pathway in erythroid cells in mice and their potential significance for human physiology and pathophysiology.