title: Identification of murine phosphodiesterase 5A isoforms and their functional characterization in HL-1 cardiac cell line.
abstract: Phosphodiesterase 5A (PDE5A) specifically degrades the ubiquitous second messenger cGMP and experimental and clinical data highlight its important role in cardiac diseases. To address PDE5A role in cardiac physiology, three splice variants of the PDE5A were cloned for the first time from mouse cDNA library (mPde5a1, mPde5a2, and mPde5a3). The predicted amino acidic sequences of the three murine isoforms are different in the N-terminal regulatory domain. mPDE5A isoforms were transfected in HEK293T cells and they showed high affinity for cGMP and similar sensitivity to sildenafil inhibition. RT-PCR analysis showed that mPde5a1, mPde5a2, and mPde5a3 had differential tissue distribution. In the adult heart, mPde5a1 and mPde5a2 were expressed at different levels whereas mPde5a3 was undetectable. Overexpression of mPDE5As induced an increase of HL-1 number cells which progress into cell cycle. mPDE5A1 and mPDE5A3 overexpression increased the number of polyploid and binucleated cells, mPDE5A3 widened HL-1 areas, and modulated hypertrophic markers more efficiently respect to the other mPDE5A isoforms. Moreover, mPDE5A isoforms had differential subcellular localization: mPDE5A1 was mainly localized in the cytoplasm, mPDE5A2 and mPDE5A3 were also nuclear localized. These results demonstrate for the first time the existence of three PDE5A isoforms in mouse and highlight their potential role in the induction of hypertrophy.
authors.affiliation: Department of Experimental Medicine, Sapienza University, Rome, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Campolo
authors.affiliation: Department of Anatomical, Histological, Forensic Medicine and Orthopedic Sciences, Sapienza University, Rome, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Zevini
authors.affiliation: Department of Biology and Biotechnology "Charles Darwin", Sapienza University, Rome, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Cardarelli
authors.affiliation: Department of Physiology and Pharmacology, Sapienza University, Rome, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Monaco
authors.affiliation: Department of Experimental Medicine, Sapienza University, Rome, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Barbagallo
authors.affiliation: Institute of Cell Biology and Neurobiology, CNR, Monterotondo, Rome, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Pellegrini
authors.affiliation: Department of Anatomical, Histological, Forensic Medicine and Orthopedic Sciences, Sapienza University, Rome, Italy.
authors.forename: Marisa
authors.initials: M
authors.lastname: Cornacchione
authors.affiliation: Department of Anatomical, Histological, Forensic Medicine and Orthopedic Sciences, Sapienza University, Rome, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Di Grazia
authors.affiliation: Department of Anatomical, Histological, Forensic Medicine and Orthopedic Sciences, Sapienza University, Rome, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: De Arcangelis
authors.affiliation: Department of Experimental Medicine, Sapienza University, Rome, Italy.
authors.forename: Daniele
authors.initials: D
authors.lastname: Gianfrilli
authors.affiliation: Department of Biology and Biotechnology "Charles Darwin", Sapienza University, Rome, Italy.
authors.forename: Mauro
authors.initials: M
authors.lastname: Giorgi
authors.affiliation: Department of Experimental Medicine, Sapienza University, Rome, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Lenzi
authors.affiliation: Department of Experimental Medicine, Sapienza University, Rome, Italy.
authors.forename: Andrea M
authors.initials: AM
authors.lastname: Isidori
authors.affiliation: Department of Anatomical, Histological, Forensic Medicine and Orthopedic Sciences, Sapienza University, Rome, Italy.
authors.forename: Fabio
authors.identifier: http://orcid.org/0000-0003-0572-5900
authors.initials: F
authors.lastname: Naro
mesh_terms: D000818:Animals; D006332:Cardiomegaly / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D002453:Cell Cycle; D002467:Cell Nucleus / Q000201:enzymology / Q000473:pathology; D049109:Cell Proliferation; D003001:Cloning, Molecular; D006152:Cyclic GMP / Q000378:metabolism; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5 / Q000235:genetics / Q000378:metabolism*; D003600:Cytosol / Q000201:enzymology; D005260:Female; D005434:Flow Cytometry; D015971:Gene Expression Regulation, Enzymologic; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D041681:NIH 3T3 Cells; D058986:Phosphodiesterase 5 Inhibitors / Q000494:pharmacology; D011123:Polyploidy; D020033:Protein Isoforms; D015398:Signal Transduction; D000068677:Sildenafil Citrate / Q000494:pharmacology; D014162:Transfection
chemical_list: D058986:Phosphodiesterase 5 Inhibitors; D020033:Protein Isoforms; D000068677:Sildenafil Citrate; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5; C517382:Pde5a protein, mouse; D006152:Cyclic GMP
keywords: PDE5A isoforms; cardiomyocytes; hypertrophy; polyploidy