Presence Of Catecholamines Research Articles

Regulation of Lysozyme Activity by Human Hormones

Lysozyme is a part of human and animal noncellular immunity. The regulation of its activity by hormones is poorly studied. The aim of this study was to test the in vitro activity of lysozyme in the presence of catecholamines, natriuretic hormones, and estradiol (E2). Hormones were incubated with lysozyme, and the activity of lysozome was further determined using a test culture of Micrococcus luteus in the early exponential growth stage. The activity of lysozyme was assessed based on the rate of change in the OD of the test culture. Molecular docking was performed using SwissDock server http://www.swissdock.ch/docking), and molecular structures were further analyzed and visualized in the UCSF Chimera 1.15rc software. According to the results, epinephrine and norepinephrine increased lysozyme activity up to 180% compared to the hormone-free enzyme. Changing the pH of the medium from 6.3 to 5.5, increased the lysozyme activity in the presence of E2 up to 150-200 %. The results also showed that exposure to hormones could modify lysozyme ctivity, and this effect depends on the temperature and pH value. The molecular docking revealed a decrease in the activation energy of the active site of enzyme during the interaction of catecholamines with the amino acid residues, asp52 and glu35 of the active site. Our findings demonstrate an additional mechanism for the involvement of lysozyme in humoral regulation of nonspecific immunity with respect to human pathogenic microflora and bacterial skin commensals by direct modulation of its activity using human hormones.

Role of Iron Phthalocyanine Coordination in Catecholamines Detection

Catecholamines are an important class of neurotransmitters responsible for regularizing, controlling, and treating neural diseases. Based on control and diseases treatment, the development of methodology and dives to sensing is a promissory technology area. This work evaluated the role of iron phthalocyanine coordination (FePc) with the specific groups from catecholamine molecules (L-dopa, dopamine, epinephrine, and the amino acid tyrosine) and the effect of this coordination on electrochemical behavior. The in situ coordination analysis was performed through isotherms π-A of FePc Langmuir films in the absence and presence of catecholamines. The π-A isotherm indicates a strong interaction between FePc monolayer and L-Dopa and DA, which present a catechol group and a side chain with a protonated amino group (-NH3+). These strong interactions with catechol and amine groups were confirmed by characterization at the molecular level using the surface-enhanced Raman spectroscopy (SERS) from a Langmuir–Schaefer monolayer deposited onto Ag surfaces. The electrochemical measurements present a similar tendency, with lower oxidation potential observed to DA>L-Dopa>Ep. The results corroborate that the coordination of the analyte on the electron mediator surface plays an essential role in an electrochemical sensing application. The FePc LS film was applied as a sensor in tablet drug samples, showing a uniformity of content of 96% for detecting active compounds present in the L-Dopa drug samples.

Interkingdom Cross-Talk in Times of Stress: Salmonella Typhimurium Grown in the Presence of Catecholamines Inhibits Porcine Immune Functionality in vitro.

In stressful situations, catecholamines modulate mammalian immune function, and in addition, they can be sensed by many bacteria. Catecholamine sensing was also found in the zoonotic gut pathogen Salmonella Typhimurium, probably contributing to the stress-induced increased risk of salmonellosis. Virulence traits such as proliferation and invasiveness are promoted upon bacterial catecholamine sensing, but it is unknown whether S. Typhimurium may also inhibit mammalian immune function in stressful situations. We thus investigated whether supernatants from S. Typhimurium grown in the presence of catecholamines modulate porcine mitogen-induced lymphocyte proliferation. Lymphocyte proliferation was reduced by supernatants from catecholamine-exposed Salmonella in a dose-dependent manner. We further examined whether adrenaline oxidation to adrenochrome, which is promoted by bacteria, could be responsible for the observed effect, but this molecule either enhanced lymphocyte functionality or had no effect. We could thereby exclude adrenochrome as a potential immunomodulating agent produced by S. Typhimurium. This study is the first to demonstrate that bacteria grown in the presence of catecholamine stress hormones alter their growth environment, probably by producing immunomodulating substances, in a way that host immune response is suppressed. These findings add a new dimension to interkingdom signaling and provide novel clues to explain the increased susceptibility of a stressed host to Salmonella infection.

Gold nanorods and their nanocomposites based on polyurethane foam for determination of catecholamines in biological fluids

Capability of gold nanorods and their nanocomposites based on polyurethane foam as spectrophotometric reagents for catecholamines determination was examined. Comparison of the analytical properties of the systems based on interaction of nanoparticles of different morphology with various reagents in the presence of catecholamines was carried out, gold nanorods-based system is characterized by the highest sensitivity. Applicability of gold nanorods and their nanocomposites for catecholamines determination in biological fluids (urine, serum) was proved by HPLC analysis. Catecholamines preconcentration by dynamic sorption on hypercrosslinked polystyrene and further elution with 6M acetic acid could serve to improve the sensitivity of the analysis.

Simple and label-free fluorescence detection of ascorbic acid in rat brain microdialysates in the presence of catecholamines

A facile and ultrasensitive sensor was constructed successfully for AA sensing based on the synergistic effect of reducing capability of AA and IFE.

Prion Peptides Are Extremely Sensitive to Copper Induced Oxidative Stress.

Copper(II) binding to prion peptides does not prevent Cu redox cycling and formation of reactive oxygen species (ROS) in the presence of reducing agents. The toxic effects of these species are exacerbated in the presence of catecholamines, indicating that dysfunction of catecholamine vesicular sequestration or recovery after synaptic release is a dangerous amplifier of Cu induced oxidative stress. Cu bound to prion peptides including the high affinity site involving histidines adjacent to the octarepeats exhibits marked catalytic activity toward dopamine and 4-methylcatechol. The resulting quinone oxidation products undergo parallel oligomerization and endogenous peptide modification yielding catechol adducts at the histidine binding ligands. These modifications add to the more common oxidation of Met and His residues produced by ROS. Derivatization of Cu-prion peptides is much faster than that undergone by Cu-β-amyloid and Cu-α-synuclein complexes in the same conditions.

A Ruthenium Complex/Carbon Nanotube Based Electrode as the First Electrochemical Sensor for Simultaneous Sensing of D‐Penicillamine, 6‐Thioguanine and Catecholamines

AbstractThe electrochemical behavior of a synthesized Ru complex (RuL) was investigated at multiwalled carbon nanotubes modified carbon paste electrode. Some special properties of the electrochemical response of this modified electrode are suitable charge transfer rate, repeatability and independance of the pH of the solution. The modified electrode can catalyze the electrochemical oxidation of penicillamine (PA) by a heterogeneous chemical reaction between PA and RuL through catalytic EC’ mechanism. The rate constant of this catalytic reaction was obtained as 1.4×106 M−1 s−1 by chronoamperometry. Because of increasing the anodic current and decreasing the oxidation potential of PA at the prepared modified electrode, this electrode was used as an electrochemical sensor for determination of PA. This sensor can determine the concentration of PA in the wide concentration range of 0.20–750.0 µM with the low detection limit of 80 nM. Simultaneous determination of PA and 6‐thioguanine in the presence of catecholamines is the other important advantage of this sensor.

Abstract 307: The Presence of an Adrenergic System in Perivascular Adipose Tissue

Perivascular adipose tissue (PVAT) is under recognized for its importance in blood pressure regulation. Visceral adipocytes reportedly contain catecholamines. Adipocytes in PVAT are directly adjacent to the blood vessels they surround and therefore the production, release and/or reuptake of catecholamines may significantly affect vascular tone. We hypothesize that an adrenergic system is present in PVAT. Glyoxylic acid staining revealed the presence of catecholamines in the cytosol of mesenteric PVAT adipocytes. Dopamine (DA), norepinephrine (NE) and epinephrine were quantified by HPLC in rat aortic PVAT, brown fat, mesenteric PVAT and retroperitoneal fat with NE being the most abundant at concentrations of 731.9, 815.0, 668.7 and 73.2 ng/g from each tissue, respectively. Two key enzymes in catecholamine synthesis; tyrosine hydroxylase and dopamine β-hydroxylase, were located to PVAT adipocytes by immunohistochemistry. The norepinephrine transporter (NET) was detected on mesenteric PVAT adipocytes by confocal microscopy using ASP+ (2μM), a fluorescent NET substrate. Uptake of ASP+ was blocked by the NET specific inhibitor nisoxetine (10μM) (see figure). These data show that PVAT contains all of the elements of an adrenergic system and may contribute to vascular and adipose function in health and disease.

Staphylococcus aureus Transporters Hts, Sir, and Sst Capture Iron Liberated from Human Transferrin by Staphyloferrin A, Staphyloferrin B, and Catecholamine Stress Hormones, Respectively, and Contribute to Virulence

Staphylococcus aureus is a frequent cause of bloodstream, respiratory tract, and skin and soft tissue infections. In the bloodstream, the iron-binding glycoprotein transferrin circulates to provide iron to cells throughout the body, but its iron-binding properties make it an important component of innate immunity. It is well established that siderophores, with their high affinity for iron, in many instances can remove iron from transferrin as a means to promote proliferation of bacterial pathogens. It is also established that catecholamine hormones can interfere with the iron-binding properties of transferrin, thus allowing infectious bacteria access to this iron pool. The present study demonstrates that S. aureus can use either of two carboxylate-type siderophores, staphyloferrin A and staphyloferrin B, via the transporters Hts and Sir, respectively, to access the transferrin iron pool. Growth of staphyloferrin-producing S. aureus in serum or in the presence of holotransferrin was not enhanced in the presence of catecholamines. However, catecholamines significantly enhanced the growth of staphyloferrin-deficient S. aureus in human serum or in the presence of human holotransferrin. It was further demonstrated that the Sst transporter was essential for this activity as well as for the utilization of bacterial catechol siderophores. The substrate binding protein SstD was shown to interact with ferrated catecholamines and catechol siderophores, with low to submicromolar affinities. Experiments involving mice challenged intravenously with wild-type S. aureus and isogenic mutants demonstrated that the combination of Hts, Sir, and Sst transport systems was required for full virulence of S. aureus.

Growth induction and stabilization of population composition in Saccharopolyspora erythraea by catecholamine compounds

Dopamine and epinephrine, hormone compounds belonging to the catecholamine group, produced a polymodal effect on the culture of the erythromycin-producing actinobacterium Saccharopolyspora erythraea. They stimulated spore germination and stabilized the composition of its population. Plating of monospore suspensions of strains RIA 1387 and RIA 120 in the presence of catecholamines increased the colony number by 2- and 2.3-fold, respectively (with epinephrine), and 3- and 4-fold, respectively (with dopamine). The optimum effect of both catecholamines was attained at a concentration of 10−5 M. The influence of exogenous catecholamines resulted in a significant shift in the population structure of the tested strains with an increased share of the colonies of the dominant morphotype, which is the most efficient antibiotic producer. Considerable differences in the biological activity of the catecholamines were revealed after exposure to ultralow temperatures both during medium-term storage (for three months) and after short-term freezing (for 10 min) in liquid nitrogen. An appreciable effect was produced by exogenous dopamine introduced at the resuscitation stage, resulting in a two- to three-fold increase in the viability of actinobacterial spores. Almost 100% of the resulting populations were composed of colonies of the dominant morphotype. In contrast to dopamine, epinephrine failed to produce a growth-stimulating effect when added to spore suspensions exposed to ultralow temperatures.

Catecholamine Interference in Enzymatic Creatinine Assays

Enzymatic creatinine assays are routinely used in clinical laboratories to provide more accurate estimated glomerular filtration rates and to avoid a perceived lack of analytical specificity associated with picrate (Jaffe) methods. Negative interferences with the enzymatic creatinine assay, which we noted in several patients on dopamine or dobutamine, prompted our further investigation into interference of catecholamines with enzymatic methods. Spiked solutions of dopamine, dobutamine, epinephrine, and norepinephrine were added to pooled sera at catecholamine concentrations consistent with clinically relevant dosing. Creatinine was measured enzymatically on the Roche P-Modular, Ortho Clinical Diagnostics Vitros 350, and Abbott i-STAT. Jaffe methods were performed on the Roche P-Modular and Siemens Dimension RxL. In 10 patients receiving dopamine and/or dobutamine via a venous or arterial line we evaluated and compared the extent of in vivo creatinine interference in paired serum samples obtained by venipuncture and from indwelling catheters. All catecholamines caused significant negative interference with the Roche enzymatic creatinine assay, most pronounced for dopamine and dobutamine. The Vitros enzymatic assay demonstrated slight negative interferences, and i-STAT enzymatic and Jaffe methods were unaffected by the presence of catecholamines. Significant (P < 0.001) differences in creatinine concentrations by Roche enzymatic vs Jaffe methods were observed in venipuncture specimens compared with arterial or venous catheter specimens, suggesting dopamine and dobutamine reversibly adhere to the catheter lumen. Negative interferences were pronounced for Roche enzymatic results in blood samples obtained from indwelling catheters, a phenomenon not observed in peripheral draws. Physicians and laboratorians should be alert to the possibility of a falsely low creatinine result and reevaluate questionable samples using a method unaffected by catecholamines.

Ca 2+ Signaling Domains Responsible For Cardiac Hypertrophy and Arrhythmias

See related articles, pages 514–521 and 522–530 Ca2+ activates and regulates multiple processes in every cell type. In the mammalian heart, cyclic fluctuations in cytosolic [Ca2+] induce and regulate the strength of cardiac contraction (termed “contractile” [Ca2+]). In addition, changes in Ca2+ appear to be centrally involved in normal and pathological signaling (termed “signaling” [Ca2+]) that regulates myocyte growth, hypertrophy, apoptosis, and necrosis.1 Whether or not contractile and signaling [Ca2+] are derived from common or distinct sources and are constrained to unique cellular microdomains is not established.2 What is clear is that cardiovascular diseases including hypertension and myocardial infarction are associated with alterations in contractile and possibly signaling [Ca2+] that are centrally involved in pathological cardiac hypertrophy, heart failure progression,1 and lethal cardiac arrhythmias.3 Defining the sources of signaling Ca2+ involved in the induction of pathological hypertrophy and the bases of dysregulated contractile [Ca2+] in cardiovascular disease should identify novel ways to treat heart disease. In this issue of Circulation Research , 2 independent reports address fundamental aspects of alterations in signaling and contractile [Ca2+]. Chiang et al4 have studied the idea that Ca2+ influx through voltage operated α1H (CaV3.2) T-type Ca2+ channels (TTCCs) is the source of the signaling [Ca2+] that activates the calcineurin (Cn)-NFAT (nuclear factor of activated T cells) signaling cascade and induces pathological cardiac hypertrophy in pressure overload. In a separate report, Terentyev et al5 explore the idea that microRNA (miR)-1, a muscle-specific microRNA that increases in abundance in cardiac disease,6 causes dysregulated contractile [Ca2+] and induces single cell arrhythmias. These 2 reports are provocative and, if independently confirmed, will have identified novel …

Free tyrosine and tyrosine-rich peptide-dependent superoxide generation catalyzed by a copper-binding, threonine-rich neurotoxic peptide derived from prion protein

Previously, generation of superoxide anion (O(2)(*-)) catalyzed by Cu-binding peptides derived from human prion protein (model sequence for helical Cu-binding motif VNITKQHTVTTTT was most active) in the presence of catecholamines and related aromatic monoamines such as phenylethylamine and tyramine, has been reported [Kawano, T., Int J Biol Sci 2007; 3: 57-63]. The peptide sequence (corresponding to helix 2) tested here is known as threonine-rich neurotoxic peptide. In the present article, the redox behaviors of aromatic monoamines, 20 amino acids and prion-derived tyrosine-rich peptide sequences were compared as putative targets of the oxidative reactions mediated with the threonine-rich prion-peptide. For detection of O(2)(*-), an O(2)(*-)-specific chemiluminescence probe, Cypridina luciferin analog was used. We found that an aromatic amino acid, tyrosine (structurally similar to tyramine) behaves as one of the best substrates for the O(2)(*-) generating reaction (conversion from hydrogen peroxide) catalyzed by Cu-bound prion helical peptide. Data suggested that phenolic moiety is required to be an active substrate while the presence of neither carboxyl group nor amino group was necessarily required. In addition to the action of free tyrosine, effect of two tyrosine-rich peptide sequences YYR and DYEDRYYRENMHR found in human prion corresponding to the tyrosine-rich region was tested as putative substrates for the threonine-rich neurotoxic peptide. YYR motif (found twice in the Y-rich region) showed 2- to 3-fold higher activity compared to free tyrosine. Comparison of Y-rich sequence consisted of 13 amino acids and its Y-to-F substitution mutant sequence revealed that the tyrosine-residues on Y-rich peptide derived from prion may contribute to the higher production of O(2)(*-). These data suggest that the tyrosine residues on prion molecules could be additional targets of the prion-mediated reactions through intra- or inter-molecular interactions. Lastly, possible mechanism of O(2)(*-) generation and the impacts of such self-redox events on the conformational changes in prion are discussed.

Neurotoxic catecholamine metabolite in nociceptors contributes to painful peripheral neuropathy

The neurotoxic effects of catecholamine metabolites have been implicated in neurodegenerative diseases. As some sensory neurons express tyrosine hydroxylase and monoamine oxidase (MAO), we investigated the potential contribution of catecholamine metabolites to neuropathic pain in a model of alcoholic neuropathy. The presence of catecholamines in sensory neurons is supported by capsaicin-stimulated epinephrine release, an effect enhanced in ethanol-fed rats. mRNA for enzymes in dorsal root ganglia involved in catecholamine uptake and metabolism, dopamine beta-hydroxylase and MAO-A, were decreased by neonatal administration of capsaicin. Ethanol-induced hyperalgesia was attenuated by systemic and local peripheral administration of inhibitors of MAO-A, reduction of norepinephrine transporter (NET) in sensory neurons and a NET inhibitor. Finally, intradermal injection of 3,4-dihydroxyphenylglycolaldehyde (DOPEGAL), a neurotoxic MAO-A catecholamine metabolite, produced robust mechanical hyperalgesia. These observations suggest that catecholamines in nociceptors are metabolized to neurotoxic products by MAO-A, which can cause neuronal dysfunction underlying neuropathic pain.

A possible mechanism of halocarbon-induced cardiac sensitization arrhythmias

Cardiac sensitization is the term used for malignant ventricular arrhythmias associated with exposure to inhaled halocarbons in the presence of catecholamines. We investigated the electrophysiological changes associated with cardiomyocyte exposure to epinephrine and a halocarbon known to be associated with cardiac sensitization (halon 1301, CF 3Br). Cardiomyocytes (CMs) were isolated from neonatal rats and grown on multielectrode arrays (MEAs). Upon exposure to epinephrine, the CM inter-spike interval (ISI) was decreased 14% at 10 μg/L ( P < 0.05) and 27% at 100 μg/L ( P < 0.05) as compared to baseline. Halon alone (50 mg/L) mildly prolonged the field potential (FP) duration (7%). CMs exposed to combinations of epinephrine (100 μg/L) and halon (50 mg/L) for 15 min showed a blunted increase in the ISI (35 ± 12%) and a 38% decrease in conduction velocity ( P < 0.05) when compared to epinephrine alone. There was no change in field potential properties, but dephosphorylated connexin 43 (Cx43) was increased 60 ± 16% with the combination as compared to epinephrine alone ( P < 0.05). Treatment with okadaic acid, a phosphatase inhibitor, prevented the Cx43 dephosphorylation and the reduction in conduction velocity upon exposure to halon and epinephrine. Moreover, the electrophysiological changes induced by epinephrine and halon were indistinguishable from those seen with the gap junction inhibitor heptanol. In conclusion, the combination of a halocarbon and epinephrine results in a unique electrophysiological signature including slow conduction that may explain, in part, the basis for cardiac sensitization. The slowing of conduction is most likely related to changes in the phosphorylation state of Cx43.

Norepinephrine Induces Lipolysis in β1/β2/β3-Adrenoceptor Knockout Mice

Catecholamines are major stimulants of adipose tissue metabolism. Norepinephrine and epinephrine act through three subtypes of beta-adrenoceptors (beta-AR) expressed in the adipocytes. The aim of this work was to study the mechanisms of lipid mobilization in beta1/beta2/beta3-AR triple-knockout (beta-less) mice. Glycerol and nonesterified fatty acids released from isolated adipocytes were measured as an index of lipolytic activity. There was no difference between the two genotypes for basal lipolysis and lipolytic response to corticotropin or to agents acting at the adenylyl cyclase and protein kinase A levels. The lipolytic response to norepinephrine and beta-AR agonists was blunted in beta-less mice. However, a residual low-affinity lipolytic effect was observed in the presence of catecholamines and beta3-AR agonists but not of beta1- or beta2-AR agonists. cAMP levels were increased by a beta-AR agonist in white and brown adipocytes of beta-less mice. The residual lipolytic effect was blocked by beta-AR antagonists. It was mediated neither by alpha1- or alpha2-AR nor dopaminergic, serotonergic, and histaminergic by receptors. Bioinformatic analyses do not provide evidence for a fourth beta-AR. We conclude that the residual lipolytic effect observed in beta-less mice can be attributed to an unknown Gs-protein-coupled receptor with low affinity for catecholamines.

Fragmentation and dimerization of copper-loaded prion protein by copper-catalysed oxidation

Prion protein consists of an N-terminal domain containing a series of octapeptide repeats with the consensus sequence PHGGGWGQ and a C-terminal domain composed of three alpha-helices and two short beta-strands. Several studies have shown that the N-terminal domain binds five Cu2+ ions. In the present study, we have investigated copper-catalysed oxidation of a recombinant mouse prion protein, PrP23-231. The copper-loaded PrP23-231 was found to be carbonylated by incubation with dopamine. Besides the formation of carbonyls, a cross-linked species with the dimeric size and C-terminally truncated species were generated. These reactions were retarded in the presence of Cu+- and Cu2+-specific copper chelators, catalase, and SOD (superoxide dismutase), but not in the presence of various bivalent metal ions. Together, these results indicate that the copper bound to prion protein undergoes catalytic cycling in the presence of catecholamines and causes the oxidation of the protein.

Type IV phosphodiesterase inhibition improves cardiac contractility in endotoxemic rats

Type IV phosphodiesterase inhibitors have a potential role in treating human sepsis. We examined the cardiac performance effects of type IV phosphodiesterase inhibition in vivo, in the absence and presence of catecholamines. Rats were randomized to receive either 4-(3-Butoxy-4-methoxybenzyl)imidazolidin-2-one (Ro 20–1724) at 0 (vehicle), 2 or 10 μg/kg/min. Utilizing a left ventricular catheter to measure cardiac performance, each animal received each of the two catecholamines, epinephrine and norepinephrine, in randomized order. Rats then received intravenous endotoxin and additional infusions of catecholamines. Ro 20–1724 at 2 μg/kg/min protected cardiac contractility during endotoxemia, and at 10 μg/kg/min increased cardiac contractility and protected cardiac function during endotoxemia. Neither dose interfered with the maximal contractile response to catecholamines. Type IV phosphodiesterase inhibition with Ro 20–1724 exerts beneficial effects on cardiac performance during septicemia in an in vivo animal model. Clinical studies of type IV phosphodiesterase inhibitors in critically ill patients are indicated.

Growth stimulation of intestinal commensal Escherichia coli by catecholamines: a possible contributory factor in trauma-induced sepsis.

Trauma is well recognized to result in the immediate and sustained release of stress-related neurochemicals such as the catecholamine norepinephrine. Past work has shown that in addition to their ability to function as neurotransmitters, catecholamines can also directly stimulate the growth of a number of pathogenic bacteria. The development of trauma-associated sepsis has often been linked to the ability of otherwise normal commensal bacteria to invade and penetrate the gut mucosal barrier. Therefore, the aim of our study was to examine whether catecholamines could also stimulate the growth of commensal Escherichia coli strains of the type present in the intestinal tract at the time of a traumatic event. Herein we report that the growth of a range of non-pathogenic isolates of E. coli of human and environmental origin was significantly increased in the presence of catecholamines. A primary mechanism by which catecholamines increase bacterial growth was shown to be iron removal from lactoferrin and transferrin and subsequent acquisition by bacteria. The 3,4-dihydroxybenzoyl (catechol) structure of the catecholamines was further demonstrated to be critical to iron acquisition. The synthetic catecholamine inotropes dobutamine and isoprenaline, as well as norepinephrine metabolites that retained the catechol structure were also active, whereas norepinephrine metabolites in which the catechol moiety had been modified were not. A role for catecholamine-mediated bacterial iron supply in the pathophysiology of gut-derived sepsis due to trauma is proposed.

Catecholamines are present in larval Xenopus laevis: A potential source for cardiac control

Changes in noradrenaline (NA), adrenaline (A), and dopamine (DA) levels in the heart, kidneys, and whole body (without heart and kidneys) during embryonic development were investigated in the frog, Xenopus laevis using high-performance liquid chromatography (HPLC). In addition, the presence of cells immunoreactive to tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH) and/or phenylethanolamine-N-methyltransferase (PNMT) in the heart of Xenopus larvae was investigated using immunohistochemical techniques. The presence of nerve fibers was visualized using antibodies against acetylated tubulin (AcT). NA and DA concentrations in the heart were low and steady in NF 40-56, showed an increased value at NF 57, and decreased again in froglets. A trend toward higher concentrations of A was observed at NF 43-49 and NF 57. Cells immunoreactive to TH, DBH, and PNMT were found in the heart from NF 40, and the TH immunoreactive cells became more abundant in the whole heart at later stages. The presence of catecholamines in the non-innervated larval heart together with the finding of TH/DBH/PNMT immunoreactive cells suggests that catecholamines are synthesized and stored in the heart and could therefore have a paracrine role in cardiac control in Xenopus larvae. Detectable concentrations of catecholamines were also found in kidneys and whole bodies (except heart and kidneys). Therefore, catecholamine-producing cells outside the heart can be an important source of circulating catecholamines involved in adrenergic cardiac control in Xenopus larvae.