In T cells, cAMP-dependent protein kinase (PKA) type I colocalizes with the cell receptor-CD3 complex (TCR/CD3) and inhibits function via a previously unknown proximal target. Here we examine mechanism for this PKA-mediated immunomodulation. cAMP treatment of Jurkat normal cells reduces Lck-mediated tyrosine phosphorylation TCR/CD3 zeta chain after activation, decreases Lck activity. Phosphorylation residue Y505 in by COOH-terminal Src (Csk), which negatively regulates Lck, is essential...

We have investigated the localization and function of epidermal growth factor receptor (EGFR) in normal cells, cholesterol-depleted cells cholesterol enriched cells. Using immunoelectron microscopy we find that EGFR is randomly distributed at plasma membrane not caveolae. Binding EGF 4°C does change EGFR,and by only small amounts bound localize to However, upon patching lipid rafts, a significant amount localized within rafts. Depletion causes increased binding EGF, dimerization EGFR,...

In this study, we report the isolation and characterization of a full-length cDNA clone for hormone-inducible regulatory subunit RIIβ (formerly called RII51) type II cAMP-dependent protein kinase from human testis library. The cloned demonstrated tissue-specific expression mRNA in tissues, with highest levels ovary.

We report benzo[b]thiophene derivatives synthesized according to a dual strategy. 8j, 9c, and 9e with affinity values toward 5-HT(7)R 5-HTT were selected probe their antidepressant activity in vivo using the forced swimming text (FST). The results showed significant after chronic treatment. 9c was effective reducing immobility time FST even acute These findings identify these compounds as new class of antidepressants rapid onset action.

Quantitative real-time RT-PCR (RT-qPCR) is a highly sensitive method for mRNA quantification, but requires invariant expression of the chosen reference gene(s). In pathological myocardium, there limited information on suitable genes other than commonly used Gapdh and 18S ribosomal RNA. Our aim was to evaluate identify in human failing rat mouse post-myocardial infarction (post-MI) heart failure across developmental stages fetal neonatal myocardium.The abundance Arbp, Rpl32, Rpl4, Tbp,...

Small molecules interfering with Rac1 activation are considered as potential drugs and already studied in animal models. A widely used inhibitor without reported attenuation of RhoA activity is NSC23766 [(N(6)-[2-[[4-(diethylamino)-1-methylbutyl]amino]-6-methyl-4-pyrimidinyl]-2-methyl-4,6-quinolinediamine trihydrochloride]. We found that inhibits the M2 muscarinic acetylcholine receptor (M2 mAChR)-induced neonatal rat cardiac myocytes. Surprisingly, NSC27366 concomitantly suppressed...

Frizzleds (FZDs) are classified as G-protein-coupling receptors, but how signals initiated and specified through heterotrimeric G proteins is unknown. FZD6 regulates convergent extension movements, its C-terminal Arg511Cys mutation causes nail dysplasia in humans. We investigated the functional relationship between FZD6, Disheveled (DVL), proteins. Live cell imaging combined with fluorescence recovery after photobleaching (FRAP) revealed that inactive human precouples to Gαi1 Gαq not...

Receptor tyrosine kinases activate mitogen-activated protein (MAP) through Ras, Raf-1, and MEK. can be transactivated by G protein-coupled receptors coupling to Gi Gq. The human serotonin 5-HT4(b) 5-HT7(a) couple Gs elevate intracellular cAMP. Certain Gs-coupled have been shown MAP a kinase A- Rap1-dependent pathway. We report the activation of extracellular signal-regulated (ERKs) 1 2 (p44 p42 kinase) in COS-7 embryonic kidney HEK293 cells. In transfected cells, 5-HT-induced ERK1/2 is...

Using membranes from stably or transiently transfected HEK293 cells cultured in 5‐HT‐free medium and expressing the recombinant human 5‐HT 7 receptor splice variants (h5‐HT 7(a) , h5‐HT 7(b) 7(d) ), we compared their abilities to constitutively activate adenylyl cyclase (AC). All elevated basal forskolin‐stimulated AC. The AC activity was reduced by antagonist methiothepin this effect blocked mesulergine (neutral antagonist) indicating that inhibitory of is inverse agonism at receptor....

We investigated the relative activity at which fully occupied human beta 1- and 2-adrenergic receptors (beta 1AR 2AR) activate stimulatory G protein (Gs)/adenylyl cyclase (AC) system in isolated membranes. The were cloned coexpressed permanent cell lines 1/beta 2 ratios that varied from 1:2 to 3:1 total receptor abundance ranged 8 2200 fmol/mg of membrane protein. Cell expressing or 2AR alone also obtained. Competitive inhibition isoproterenol-stimulated AC by 2-selective antagonist ICI...

Levosimendan is known as a calcium sensitizer, although it also to inhibit PDE3. We aimed isolate each component and estimate their contribution the increased cardiac contractility induced by levosimendan.Contractile force was measured in electrically stimulated ventricular strips from explanted failing human hearts left normal male Wistar rats. PDE activity two-step assay on ventricle.Levosimendan exerted positive inotropic effect (PIE) reaching maximum at 10(-5) M hearts. In presence of...

The cDNAs encoding the murine LH receptor (LHR) and human beta 2-adrenoceptor (h 2AR) were cloned RNAs complementary to their sense strands (cRNAs) injected into defolliculated Xenopus oocytes. This led expression, respectively, of LH- isoproterenol-stimulable adenylyl cyclase activities, indicating that functionally active had been cloned. In oocytes with LHR cRNA, but not in control or h 2AR cRNA-injected oocytes, CG increased a Ca(2+)-activated Cl- current, as measured by...

We report the molecular cloning of a fragment human genomic DNA called 512, containing an open reading frame 1170 nucleotides, which encodes receptor for serotonin 390 amino acids.The function S12 protein was demonstrated by functional expression in mouse LS12 cells obtained stable transfection Ltk-cells, and LMSS12 cells, derived from LM6 (Ltkcells previously transfected with MS muscarinic acetylcholine receptor).Adenylyl cyclase studies showed that 512 is able to mediate inhibition...

In this report we demonstrate glucocorticoid receptors in seminiferous tubules of the rat testis, and that these are localized Sertoli cells peritubular cells. The had high affinity for [3H]dexamethasone (Kd = 0.5 - 1 x 10(-9) M), similar Kd values were calculated from equilibrium analysis rate studies (k1 1.5 10(6) M-1 min-1 k-1 1.4 10(-3) min-1, O C). Binding specificity was typical (affinity: dexamethasone greater than corticosterone cortisol approximately R5020 progesterone aldosterone...

Whereas natriuretic peptides increase cGMP levels with beneficial cardiovascular effects through protein kinase G, we found an unexpected cardio-excitatory effect of C-type peptide (CNP) receptor B (NPR-B) stimulation in failing cardiac muscle and explored the mechanism. Heart failure was induced male Wistar rats by coronary artery ligation. Contraction studies were performed left ventricular strips. Cyclic nucleotides measured radio- enzyme immunoassay. Apoptosis determined isolated...

Background and Purpose PDE3 and/or PDE4 control ventricular effects of catecholamines in several species but their relative failing human ventricle are unknown. We investigated whether the ‐selective inhibitor cilostamide (0.3–1 μM) or rolipram (1–10 modified positive inotropic lusitropic myocardium. Experimental Approach Right left trabeculae from freshly explanted hearts 5 non‐β‐blocker‐treated 15 metoprolol‐treated patients with terminal heart failure were paced to contract at 1 Hz. The...

Background All-trans retinoic acid (atRA), an active derivative of vitamin A, regulates cell differentiation, proliferation and cardiac morphogenesis via transcriptional activation receptors (RARs) acting on response elements (RARE).We hypothesized that the (RA) signalling pathway is activated in myocardial ischemia postischemic remodelling. Methods Findings Myocardial infarction was induced through ligating left coronary artery mice. In vivo RARs measured by imaging RARE-luciferase reporter...