Redox sensitivity of tyrosine hydroxylase activity and expression in dopaminergic dysfunction

Oxidant molecules generated during neuronal metabolism appear to play a significant role in the processes of aging and neurodegeneration. Increasing experimental evidence suggests the noteworthy relevance of the intracellular reduction-oxidation (redox) balance for the dopaminergic (DA-ergic) neurons of the substantia nigra pars compacta. These cells possess a distinct physiology intrinsically associated with elevated reactive oxygen species production, conferring on them a high vulnerability to free radical damage, one of the major causes of selective DA-ergic neuron dysfunction and degeneration related to neurological disorders such as Parkinson’s disease. Tyrosine hydroxylase (tyrosine 3-monooxygenase; E.C. 1.14.16.2; TH) activity represents the rate-limiting biochemical event in DA synthesis. TH activity, metabolism and expression are finely tuned by several regulatory systems in order to maintain a crucial physiological condition in which DA synthesis is closely coupled to its secretion. Alterations of these regulatory systems of TH functions have indeed been thought to be key events in the DA-ergic degeneration. TH has seven cysteine residues presenting thiols. Depending on the oxido-reductive (redox) status of the cellular environment, thiols exist either in the reduced form of free thiols or oxidized to disulfides. The formation of disulfides in proteins exerts critical regulatory functions both in physiological and in pathological conditions when oxidative stress is sustained. Several reports have recently shown that redox state changes of thiol residues, as consequence of an oxidative injury, can directly or indirectly affect the TH activity, metabolism and expression. The major focus of this review, therefore, is to report recent evidence on the redox modulation of TH activity and expression, and to provide an overview of a cellular phenomenon that might represent a target for new therapeutic strategies against the DA-ergic neurodegenerative disorders.peer-reviewe

Steady-State Kinetics of α-Synuclein Ferrireductase Activity Identifies the Catalytically Competent Species

α-Synuclein (α-syn) is a cytosolic protein known for its association with neurodegenerative diseases, including Parkinson’s disease and other synucleinopathies. The potential cellular function of α-synuclein may be of consequence for understanding the pathogenesis of such diseases. Previous work has suggested that α-synuclein can catalyze the reduction of iron as a ferrireductase. We performed a detailed analysis of the steady-state kinetics of recombinant α-syn ferrireductase activity and for disease-associated variants. Our study illustrates that the ferrireductase activity we observed is clearly commensurate with bona fide enzyme activity and suggests a mechanistic rationale for the activity and the relationship to cellular regulation of the pool of Fe­(III) and Fe­(II). Using cell-based studies, we examined the functionally active conformation and found that the major catalytically active form is a putative membrane-associated tetramer. Using an artificial membrane environment with recombinant protein, we demonstrate that secondary structure folding of α-synuclein is insufficient to allow enzyme activity and the absolute specificity of the tertiary/quaternary structure is the primary requirement. Finally, we explored the steady-state kinetics of a range of disease α-synuclein variants and found that variants involved in neurodegenerative disease exhibited major changes in their enzymatic activity. We discuss these data in the context of a potential disease-associated mechanism for aberrant α-synuclein ferrireductase activity

Homeostatic mechanisms in dopamine synthesis and release: a mathematical model

<p>Abstract</p> <p>Background</p> <p>Dopamine is a catecholamine that is used as a neurotransmitter both in the periphery and in the central nervous system. Dysfunction in various dopaminergic systems is known to be associated with various disorders, including schizophrenia, Parkinson's disease, and Tourette's syndrome. Furthermore, microdialysis studies have shown that addictive drugs increase extracellular dopamine and brain imaging has shown a correlation between euphoria and psycho-stimulant-induced increases in extracellular dopamine <abbrgrp><abbr bid="B1">1</abbr></abbrgrp>. These consequences of dopamine dysfunction indicate the importance of maintaining dopamine functionality through homeostatic mechanisms that have been attributed to the delicate balance between synthesis, storage, release, metabolism, and reuptake.</p> <p>Methods</p> <p>We construct a mathematical model of dopamine synthesis, release, and reuptake and use it to study homeostasis in single dopaminergic neuron terminals. We investigate the substrate inhibition of tyrosine hydroxylase by tyrosine, the consequences of the rapid uptake of extracellular dopamine by the dopamine transporters, and the effects of the autoreceoptors on dopaminergic function. The main focus is to understand the regulation and control of synthesis and release and to explicate and interpret experimental findings.</p> <p>Results</p> <p>We show that the substrate inhibition of tyrosine hydroxylase by tyrosine stabilizes cytosolic and vesicular dopamine against changes in tyrosine availability due to meals. We find that the autoreceptors dampen the fluctuations in extracellular dopamine caused by changes in tyrosine hydroxylase expression and changes in the rate of firing. We show that short bursts of action potentials create significant dopamine signals against the background of tonic firing. We explain the observed time courses of extracellular dopamine responses to stimulation in wild type mice and mice that have genetically altered dopamine transporter densities and the observed half-lives of extracellular dopamine under various treatment protocols.</p> <p>Conclusion</p> <p>Dopaminergic systems must respond robustly to important biological signals such as bursts, while at the same time maintaining homeostasis in the face of normal biological fluctuations in inputs, expression levels, and firing rates. This is accomplished through the cooperative effect of many different homeostatic mechanisms including special properties of tyrosine hydroxylase, the dopamine transporters, and the dopamine autoreceptors.</p

CHRONIC LOW-LEVEL LEAD EXPOSURE AFFECTS THE MONOAMINERGIC SYSTEM IN THE MOUSE SUPERIOR OLIVARY COMPLEX

Low-level lead (Pb) exposure is associated with behavioral and cognitive dysfunction. It is not clear how Pb produces these behavioral changes but low-level Pb exposure and learning disabilities have been associated with altered auditory temporal processing in both humans and animals. Temporal processing is used to decode complex sounds and to detect a signal within a noise background, and it is thought that neurons of the superior olivary complex (SOC) in the brainstem play a role in sound detection in noisy environments and in selective auditory attention. The SOC receives a catecholaminergic and a serotonergic innervation from the locus coeruleus and the dorsal raphe respectively. While the physiological role of the noradrenergic input has yet to be defined, serotonin is involved in auditory temporal processing. Because Pb exposure modulates auditory temporal processing, the serotonergic system is a potential target for Pb. The current study was undertaken to determine whether developmental Pb exposure preferentially changes the expression of serotonin within the SOC. Pb-treated mice were exposed to no Pb, 0.01 mM (very low) or 0.1 mM (Low) Pb acetate throughout gestation and through 21 days postnatally. Brainstem sections from control and Pb-exposed mice were immunostained for the vesicular monoamine transporter 2 (VMAT2), serotonin, and dopamine beta hydroxylase (DβH, a marker for norepinephrine) in order to elucidate the effect of Pb on monoaminergic input into the SOC. In addition, sections were immunolabeled with antibodies to VGLUT1, VGAT and VAChT in order to determine whether Pb exposure alters the glutaminergic, gaba-ergic, or cholinergic systems. Pb exposure caused a significant decrease in VMAT2, 5HT, and DβH expression while VGLUT1, VGAT and VAChT showed no change. These results provide evidence that Pb exposure during development alters normal monoaminergic expression in the auditory brainstem

To Do or Not to Do: Dopamine, Affordability and the Economics of Opportunity

Five years ago, we introduced the thrift hypothesis of dopamine (DA), suggesting that the primary role of DA in adaptive behavior is regulating behavioral energy expenditure to match the prevailing economic conditions of the environment. Here we elaborate that hypothesis with several new ideas. First, we introduce the concept of affordability, suggesting that costs must necessarily be evaluated with respect to the availability of resources to the organism, which computes a value not only for the potential reward opportunity, but also the value of resources expended. Placing both costs and benefits within the context of the larger economy in which the animal is functioning requires consideration of the different timescales against which to compute resource availability, or average reward rate. Appropriate windows of computation for tracking resources requires corresponding neural substrates that operate on these different timescales. In discussing temporal patterns of DA signaling, we focus on a neglected form of DA plasticity and adaptation, changes in the physical substrate of the DA system itself, such as up- and down-regulation of receptors or release probability. We argue that changes in the DA substrate itself fundamentally alter its computational function, which we propose mediates adaptations to longer temporal horizons and economic conditions. In developing our hypothesis, we focus on DA D2 receptors (D2R), arguing that D2R implements a form of “cost control” in response to the environmental economy, serving as the “brain’s comptroller”. We propose that the balance between the direct and indirect pathway, regulated by relative expression of D1 and D2 DA receptors, implements affordability. Finally, as we review data, we discuss limitations in current approaches that impede fully investigating the proposed hypothesis and highlight alternative, more semi-naturalistic strategies more conducive to neuroeconomic investigations on the role of DA in adaptive behavior

Einfluss von Lichtspektren auf wertgebende Inhaltsstoffe und Qualität asiatischer Gemüsepflanzen, am Beispiel von Persicaria odorata (Lour.) Sojak

In der vorliegenden Arbeit wurde der Einfluss von Lichtbedingungen auf den Ertrag und die nutritive Qualität der Versuchspflanze Persicaria odorata untersucht. Die Experimente wurden unter praxisnahen Bedingungen im Gewächshaus (Sonnenlicht) und Klimakammern (Dreiband-Leuchtstoffröhren) unter Berücksichtigung eines breiten Inhaltsstoffspektrums, mehreren Versuchsreihen, Lichtmodifizierungen anhand von Zusatzbestrahlungen mit LEDs und Überdeckungen mit photoselektiven Folien sowie dem Anwenden verschiedener statistischer Auswertungsverfahren durchgeführt. Die Tageslichtsumme beeinflusste das Pflanzenwachstum positiv. Beispielsweise wurde die Frischmasse bei einem Anstieg der Tageslichtsumme um 1% mit gleichbleibender spektralen Lichtzusammensetzung um 0,96% (Gewächshaus) bzw. 0,64% (Klimakammer) erhöht. Versuche mit monochromatischen LEDs zeigten, dass spektralbereichsabhängige Erhöhungen der Tageslichtsumme die Biomassesteigerungen verstärkten. Den deutlichsten Effekt im Hinblick auf die Frischmassebildung hatte das kurzwellige Blaulicht (443 nm). Bezüglich der Inhaltsstoffe zeigte insbesondere das kurzwellige Grünlicht (515 nm) einen positiven Effekt. Dieser Spektralbereich wird jedoch durch die traditionell in Gewächshäusern eingesetzten Natriumhochdrucklampen kaum emittiert. Auch sind die bisher häufig eingesetzten Bestrahlungen mit Blau- und Rotlicht in kontrollierten Anbausystemen zwar wichtig für den Aufbau von Biomasse, jedoch nicht optimal im Hinblick auf den Nährwert. Die Ergebnisse zeigen, dass die Belichtung im Gartenbau sorgfältig ausgewählt werden sollte, jedoch während strahlungsschwachen Monaten eine effektive Möglichkeit ist, den Ertrag und die nutritive Qualität von P. odorata deutlich erhöhten. Durch Modifikationen der Lichtbedingungen ist es möglich den Ertrag und Nährwert pflanzlicher Erzeugnisse zu steigern und dadurch die Versorgungslage mit gesundheitsfördernden bzw. essentiellen pflanzlichen Inhaltsstoffen zu verbessern.This thesis examined the impact of light conditions on the nutritive quality of the test plant Persicaria odorata. Experiments took place in a greenhouse (sunlight) and climate chamber (fluorescent tube) and under consideration of a wide range of nutritive compounds, several test series, different light modifications and the use of various statistically evaluation procedures. This thesis demonstrates that varying light conditions increase nutritive value and yield under realistic conditions. Increasing the day light integral by 1% improved the fresh mass of 0,96% (greenhouse) and 0,64% (climate chamber). Experiments with monochromatic LEDs proved a high increase of biomass by spectral range dependent enhancements of the day light integral. For example, in the greenhouse, fresh mass was mostly affected by short wave blue radiation (443 nm). Additionally, the day light integral influenced the accumulation of nutrients. Higher contents of polyphenols and flavonoids in the climate chamber and the reactions on additional monochromatic LEDs proved again that it is more effective to increase day light integral by a specific enhancement of certain spectral ranges. The highest increases were achieved by supplementing spectral ranges that are not included in the emission spectrum of commonly used high pressure sodium lamps. Results also indicate that controlled agriculture’s frequently used lighting systems which emit blue and red light can improve plant growth, but it is not optimal regarding the nutritive quality. Adding short wave green light (515 nm) could increase biomass and improve the nutritive quality considerably. This thesis demonstrates that additional lighting in horticulture is an effective method to improve yield and nutritive quality of herbal vegetables in light deprived northern regions. By altering the emission spectrum of traditional artificial lighting systems, it is possible to increase yield and simultaneously improve the nutritive value of vegetables