title: Role of Nitric Oxide, Nitric Oxide Synthase, Soluble Guanylyl Cyclase, and cGMP-Dependent Protein Kinase I in Mouse Stem Cell Cardiac Development.
abstract: Introduction and Aim. Nitric oxide (NO) can trigger cardiac differentiation of embryonic stem cells (ESCs), indicating a cardiogenic function of the NO synthetizing enzyme(s) (NOS). However, the involvement of the NO/NOS downstream effectors soluble guanylyl cyclase (sGC) and cGMP activated protein kinase I (PKG-I) is less defined. Therefore, we assess the involvement of the entire NO/NOS/sGC/PKG-I pathway during cardiac differentiation process.Methods. Mouse ESCs were differentiated toward cardiac lineages by hanging drop methodology for 21 days. NOS/sGC/PKG-I pathway was studied quantifying genes, proteins, enzymatic activities, and effects of inhibition during differentiation. Percentages of beating embryoid bodies (mEBs) were evaluated as an index of cardiogenesis.Results and Discussion. Genes and protein expression of enzymes were increased during differentiation with distinctive kinetics and proteins possessed their enzymatic functions. Exogenous administered NO accelerated whereas the blockade of PKG-I strongly slowed cardiogenesis. sGC inhibition was effective only at early stages and NOS blockade ineffective. Of NOS/sGC/PKG-I pathway, PKG-I seems to play the prominent role in cardiac maturation.Conclusion. We concluded that exogenous administered NO and other pharmacological strategies able to increase the activity of PKG-I provide new tools to investigate and promote differentiation of cardiogenic precursors.
authors.affiliation: Department of Neurofarba, Pharmacology and Toxicology Unit, University of Florence, Florence, Italy; Interuniversity Center for Molecular Medicine and Applied Biophysics, Florence, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Spinelli
authors.affiliation: Department of Neurofarba, Pharmacology and Toxicology Unit, University of Florence, Florence, Italy.
authors.forename: Alessia
authors.initials: A
authors.lastname: Vona
authors.affiliation: Department of Neurofarba, Pharmacology and Toxicology Unit, University of Florence, Florence, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Corti
authors.affiliation: Department of Neurofarba, Pharmacology and Toxicology Unit, University of Florence, Florence, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Diolaiuti
authors.affiliation: Department of Neurofarba, Pharmacology and Toxicology Unit, University of Florence, Florence, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Zanardelli
authors.affiliation: Department of Neurofarba, Pharmacology and Toxicology Unit, University of Florence, Florence, Italy; Interuniversity Center for Molecular Medicine and Applied Biophysics, Florence, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Sartiani
authors.affiliation: Department of Neurofarba, Pharmacology and Toxicology Unit, University of Florence, Florence, Italy.
authors.forename: Paola
authors.identifier: 0000-0003-3879-9068
authors.initials: P
authors.lastname: Failli