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Identification of novel cyclic nucleotide phosphodiesterase gene cDNAs in the brain of guinea pig.

ABSTRACT: Cyclic nucleotide phosphodiesterase (PDE) is a critical component of the nitric oxide (NO) signaling pathway and plays critical roles in cognition and learning, Parkinson's disease, attention deficit hyperactivity disorder, psychosis and depression. The PDEs in the brain of guinea pig have not yet been reported. The present study aimed to detect the unknown Pde cDNAs in the brain of guinea pig.Reverse transcription polymerase chain reaction (RT-PCR) and sequence comparison analysis were performed to detect the expression of Pde cDNAs and to assess the identity rates of cDNA and amino acid sequences between guinea pig and human or mouse, respectvely. The RT-PCR primers were located on the conserved region of human PDE and mouse Pde cDNAs.Eleven novel Pde cDNAs were detected in the brain of guinea pig (Cavia porcellus), including CpPde1a, CpPde1b, CpPde2a, CpPde4a, CpPde4d, CpPde5a, CpPde6c, CpPde7b, CpPde8a, CpPde9a, and CpPde10a. The identity rates of the Pde cDNA sequences between guinea pig and human ranged from 83.8% to 94.3%, and those of the amino acid sequences ranged from 91.9% to 100%. The identity rates of Pde cDNA sequences between guinea pig and mouse ranged from 84.6% to 92.1%, and those of amino acid sequences ranged from 91.2% to 99.2%. The average identity rate of the 11 Pde cDNA sequences between guinea pig and human was significantly higher (P < 0.01) than that between guinea pig and mouse. The putative partial amino acid sequences of guinea pig contained at least one of the conserved domains of human and mouse PDE proteins.These results indicate that the brain-expressed Pde genes are identified in guinea pig, which lays the foundation for further investigating the physiological roles of PDE proteins in the brain.

SUBMITTER: Long YS

PROVIDER: S-EPMC5560350 | biostudies-other | 2010 Oct

REPOSITORIES: biostudies-other

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Project description:BackgroundHeart failure is a leading cause of death worldwide. Cyclic nucleotide phosphodiesterases (PDEs), through degradation of cyclic nucleotides, play critical roles in cardiovascular biology and disease. Our preliminary screening studies have revealed PDE10A upregulation in the diseased heart. However, the roles of PDE10A in cardiovascular biology and disease are largely uncharacterized. The current study is aimed to investigate the regulation and function of PDE10A in cardiac cells and in the progression of cardiac remodeling and dysfunction.MethodsWe used isolated adult mouse cardiac myocytes and fibroblasts, as well as preclinical mouse models of hypertrophy and heart failure. The PDE10A selective inhibitor TP-10, and global PDE10A knock out mice were used.ResultsWe found that PDE10A expression remains relatively low in normal and exercised heart tissues. However, PDE10A is significantly upregulated in mouse and human failing hearts. In vitro, PDE10A deficiency or inhibiting PDE10A with selective inhibitor TP-10, attenuated cardiac myocyte pathological hypertrophy induced by Angiotensin II, phenylephrine, and isoproterenol, but did not affect cardiac myocyte physiological hypertrophy induced by IGF-1 (insulin-like growth factor 1). TP-10 also reduced TGF-? (transforming growth factor-?)-stimulated cardiac fibroblast activation, proliferation, migration and extracellular matrix synthesis. TP-10 treatment elevated both cAMP and cGMP levels in cardiac myocytes and cardiac fibroblasts, consistent with PDE10A as a cAMP/cGMP dual-specific PDE. In vivo, global PDE10A deficiency significantly attenuated myocardial hypertrophy, cardiac fibrosis, and dysfunction induced by chronic pressure overload via transverse aorta constriction or chronic neurohormonal stimulation via Angiotensin II infusion. Importantly, we demonstrated that the pharmacological effect of TP-10 is specifically through PDE10A inhibition. In addition, TP-10 is able to reverse pre-established cardiac hypertrophy and dysfunction. RNA-Sequencing and bioinformatics analysis further identified a PDE10A-regualted transcriptome involved in cardiac hypertrophy, fibrosis, and cardiomyopathy.ConclusionsTaken together, our study elucidates a novel role for PDE10A in the regulation of pathological cardiac remodeling and development of heart failure. Given that PDE10A has been proven to be a safe drug target, PDE10A inhibition may represent a novel therapeutic strategy for preventing and treating cardiac diseases associated with cardiac remodeling.

Project description:Trypanosoma cruzi, the causative agent of Chagas disease, encodes a number of different cAMP-specific PDE (phosphodiesterase) families. Here we report the identification and characterization of TcrPDEB1 and its comparison with the previously identified TcrPDEB2 (formerly known as TcPDE1). These are two different PDE enzymes of the TcrPDEB family, named in accordance with the recent recommendations of the Nomenclature Committee for Kinetoplast PDEs [Kunz, Beavo, D'Angelo, Flawia, Francis, Johner, Laxman, Oberholzer, Rascon, Shakur et al. (2006) Mol. Biochem. Parasitol. 145, 133-135]. Both enzymes show resistance to inhibition by many mammalian PDE inhibitors, and those that do inhibit do so with appreciable differences in their inhibitor profiles for the two enzymes. Both enzymes contain two GAF (cGMP-specific and -stimulated phosphodiesterases, Anabaena adenylate cyclases and Escherichia coli FhlA) domains and a catalytic domain highly homologous with that of the T. brucei TbPDE2/TbrPDEB2 family. The N-terminus+GAF-A domains of both enzymes showed significant differences in their affinities for cyclic nucleotide binding. Using a calorimetric technique that allows accurate measurements of low-affinity binding sites, the TcrPDEB2 N-terminus+GAF-A domain was found to bind cAMP with an affinity of approximately 500 nM. The TcrPDEB1 N-terminus+GAF-A domain bound cAMP with a slightly lower affinity of approximately 1 muM. The N-terminus+GAF-A domain of TcrPDEB1 did not bind cGMP, whereas the N-terminus+GAF-A domain of TcrPDEB2 bound cGMP with a low affinity of approximately 3 muM. GAF domains homologous with those found in these proteins were also identified in related trypanosomatid parasites. Finally, a fluorescent cAMP analogue, MANT-cAMP [2'-O-(N-methylanthraniloyl)adenosine-3',5'-cyclic monophosphate], was found to be a substrate for the TcPDEB1 catalytic domain, opening the possibility of using this molecule as a substrate in non-radioactive, fluorescence-based PDE assays, including screening for trypanosome PDE inhibitors.

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Identification of novel cyclic nucleotide phosphodiesterase gene cDNAs in the brain of guinea pig.

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