Nadine Saul

Hormetins, antioxidants and prooxidants: defining quercetin-, caffeic acid- and rosmarinic acid-mediated life extension in C. elegans

Quercetin, Caffeic- and Rosmarinic acid exposure extend lifespan in Caenorhabditis elegans. This comparative study uncovers basic common and contrasting underlying mechanisms: For all three compounds, life extension was characterized by hormetic dose response curves, but hsp-level expression was variable. Quercetin and Rosmarinic acid both suppressed bacterial growth; however, antibacterial properties were not the dominant reason for life extension. Exposure to Quercetin, Caffeic- and Rosmarinic acid resulted in reduced body size, altered lipid-metabolism and a tendency towards a delay in reproductive timing; however the total number of offspring was not affected. An indirect dietary restriction effect, provoked by either chemo-repulsion or diminished pharyngeal pumping was rejected. Quercetin and Caffeic acid were shown to increase the antioxidative capacity in vivo and, by means of a lipofuscin assay, reduce the oxidative damage in the nematodes. Finally, it was possible to demonstrate that the life and thermotolerance enhancing properties of Caffeic- and Rosmarinic acid both rely on osr-1, sek-1, sir-2.1 and unc-43 plus daf-16 in the case of Caffeic acid. Taken together, hormesis, in vivo antioxidative/prooxidative properties, modulation of genetic players, as well as the re-allocation of energy all contribute (to some extent and dependent on the polyphenol) to life extension.

Quercetin mediated lifespan extension in Caenorhabditis elegans is modulated by age-1, daf-2, sek-1 and unc-43

The nematode Caenorhabditis elegans responds to flavonoid-rich diets with improved health and longevity. The precise mechanism(s) responsible for this remains to be identified, but is believed to be linked to the highly antioxidative properties of flavonoids. This study provides a dissection of lifespan modulation by the flavonoid quercetin. In detail, quercetin was shown not to act as a simple antimicrobial agent or exclusively via radical scavenging capacities. Likewise, lifespan extension had no effect on reproduction and body length. Furthermore, neither a caloric restriction mimetic nor a sirtuin (sir-2.1) dependence was identified as a likely mode of action. However, four genes were pinpointed to be required for the quercetin derived lifespan extension, namely age-1, daf-2, unc-43 and sek-1. The latter two have, to date, not been linked to quercetin-mediated lifespan extension.

Quercetin-mediated longevity in Caenorhabditis elegans : Is DAF-16 involved?

The polyphenol quercetin has recently been found to extend lifespan and increase stress resistance in the nematode Caenorhabditis elegans. The forkhead transcription factor DAF-16 has previously been linked to these effects. However, by using a daf-16(mgDf50) mutant strain, we show that quercetin exposure leads to increased mean lifespans up to 15%. Furthermore, quercetin-treated daf-16(mgDf50) worms show an enhanced resistance to thermal and oxidative stress. Our data reveal that DAF-16 is not obligatorily required for quercetin-mediated longevity and stress resistance.

Meta-analysis of global transcriptomics suggests that conserved genetic pathways are responsible for Quercetin and Tannic acid mediated longevity in C. elegans

Recent research has highlighted that the polyphenols Quercetin and Tannic acid are capable of extending the lifespan of Caenorhabditis elegans. To gain a deep understanding of the underlying molecular genetics, we analyzed the global transcriptional patterns of nematodes exposed to three concentrations of Quercetin or Tannic acid, respectively. By means of an intricate meta-analysis it was possible to compare the transcriptomes of polyphenol exposure to recently published datasets derived from (i) longevity mutants or (ii) infection. This detailed comparative in silico analysis facilitated the identification of compound specific and overlapping transcriptional profiles and allowed the prediction of putative mechanistic models of Quercetin and Tannic acid mediated longevity. Lifespan extension due to Quercetin was predominantly driven by the metabolome, TGF-beta signaling, Insulin-like signaling, and the p38 MAPK pathway and Tannic acid’s impact involved, in part, the amino acid metabolism and was modulated by the TGF-beta and the p38 MAPK pathways. DAF-12, which integrates TGF-beta and Insulin-like downstream signaling, and genetic players of the p38 MAPK pathway therefore seem to be crucial regulators for both polyphenols. Taken together, this study underlines how meta-analyses can provide an insight of molecular events that go beyond the traditional categorization into gene ontology-terms and Kyoto encyclopedia of genes and genomes-pathways. It also supports the call to expand the generation of comparative and integrative databases, an effort that is currently still in its infancy.

The non-target organism Caenorhabditis elegans withstands the impact of sulfamethoxazole

The widespread usage of antibiotics in agriculture leads to releases into the environment, but there is insufficient knowledge of the side-effects on non-target organisms. Therefore, we investigated the effects of the sulfonamide-antibiotic sulfamethoxazole (SMX) on Caenorhabditis elegans at phenotypic, biochemical and molecular biological levels. Multiple endpoints, including life history traits, thermal stress resistance and lipid peroxidation, as well as gene expression profiles, were determined after exposure of the nematodes to SMX. In contrast to expectations, SMX prolonged the lifespan and increased both the body size and pharynx pumping rate. On the other hand, SMX delayed reproductive timing and caused lipid peroxidation. The total number of offspring and thermal stress resistance were unaffected. The up-regulation of hsp-16.1 indicated stress in general and the increased lipid peroxidation oxidative stress in particular. This oxidative stress indicated that mitohormesis was the likely cause of the longevity and that enhanced pumping frequency was probably the reason for the increased growth. The sole adverse effect was delayed initial reproduction. This delay, however, can be crucial for r-strategists, such as the bacterivorous model animal used, in sustaining their populations in the environment in the presence of predators. Bacterivorous animals, in turn, are essential to maintaining nutrient recycling via the microbial loop.

Interaction of temperature and an environmental stressor: Moina macrocopa responds with increased body size, increased lifespan, and increased offspring numbers slightly above its temperature optimum

For organisms, temperature is one of the most important environmental factors and gains increasing importance due to global warming, since increasing temperatures may pose organisms close to their environmental tolerance limits and, thus, they may become more vulnerable to environmental stressors. We analyzed the temperature-dependence of the water-soluble antioxidant capacity of the cladoceran Moina macrocopa and evaluated its life trait variables with temperature (15, 20, 25, 30°C) and humic substance (HS) concentrations (0, 0.18, 0.36, 0.90, 1.79 mM DOC) as stressors. Temperatures below and above the apparent optimum (20°C) reduced the antioxidative capacity. Additions of HSs increased body length, but decreased mean lifespan at 15 and 20°C. There was no clear HS-effect on offspring numbers at 15, 20, and 30°C. At 25°C with increasing HS-concentration, lifespan was extended and offspring numbers increased tremendously, reaching 250% of the control. Although the applied HS preparation possesses estrogenic and antiandrogenic activities, a xenohormone mechanism does not seem plausible for the reproductive increase, because comparable effects did not occur at other temperatures. A more convincing explanation appears to be the mitohormesis hypothesis which states that a certain increase of reactive oxygen production leads to improved health and longevity and, with Moina, also to increased offspring numbers. Our results suggest that at least with the eurythermic M. macrocopa, a temperature above the optimum can be beneficial for several life trait variables, even when combined with a chemical stressor. Temperatures approximately 10°C above its optimum appear to adversely affect the lifespan and reproduction of M. macrocopa. This indicates that this cladoceran species seems to be able to utilize temperature as an ecological resource in a range slightly above its thermal optimum.

UV-induced DNA damage in Cyclops abyssorum tatricus populations from clear and turbid alpine lakes

Zooplankton from clear alpine lakes thrive under high levels of solar UV radiation (UVR), but in glacially turbid ones they are more protected from this damaging radiation. Here, we present results from experiments done with Cyclops abyssorum tatricus to assess UV-induced DNA damage and repair processes using the comet assay. Copepods were collected from three alpine lakes of differing UV transparency ranging from clear to glacially turbid, and exposed to artificial UVR. In addition, photoprotection levels [mycosporine-like amino acids (MAAs) and lipophilic antioxidant capacity] were estimated in the test populations. Similar UV-induced DNA damage levels were observed among the copepods from all lakes, but background DNA damage (time zero and dark controls) was lowest in the copepods from the glacially turbid lake, resulting in a higher relative DNA damage accumulation. Most DNA strand breaks were repaired after recovery in the dark. Low MAA concentrations were found in the copepods from the glacially turbid lake, while the highest levels were observed in the population from the most UV transparent lake. However, the highest lipophilic antioxidant capacities were measured in the copepods from the lake with intermediate UV transparency. Photoprotection and the ability to repair DNA damage, and consequently reducing UV-induced damage, are part of the response mechanisms in zooplankton to changes in water transparency caused by glacier retreat.

Hormesis and longevity with tannins: Free of charge or cost-intensive?

Hormetic lifespan extension is, for obvious reasons, beneficial to an individual. But is this effect really cost-neutral? To answer this question, four tannic polyphenols were tested on the nematode Caenorhabditis elegans. All were able to extend the lifespan, but only some in a hormetic fashion. Additional life trait variables including stress resistance, reproductive behavior, growth, and physical fitness were observed during the exposure to the most life extending concentrations. These traits represent the quality of life and the population fitness, being the most important parameters of a hormetic treatment besides lifespan. Indeed, it emerged that each life-extension is accompanied by a constraining effect in at least one other endpoint, for example growth, mobility, stress resistance, or reproduction. Thus, in this context, longevity could not be considered to be attained for free and therefore it is likely that other hormetic benefits may also incur cost-intensive and unpredictable side-effects.

Leaf litter leachates have the potential to increase lifespan, body size, and offspring numbers in a clone of Moina macrocopa.

Leaf litter processing is one major pathway of the global organic carbon cycle. During this process, a variety of small reactive organic compounds are released and transported to the aquatic environment, and may directly impact aquatic organisms as natural xenobiotics. We hypothesize that different forest stockings produce different leachate qualities, which in turn, stress the aquatic communities and, eventually, separate sensitive from tolerant species. Particularly, leachates from coniferous trees are suspected to have strongly adverse impacts on sensitive species. We exposed individuals of a clone of the model organism, Moina macrocopa, to comparable concentrations (approximately 2mM) of litter leachates of Norway spruce, Picea abies, Colorado blue spruce, Picea pungens, black poplar, Populus nigra, and sessile oak, Quercus petraea. The animals were fed ad libitum. The following life trait variables were recorded: growth, lifespan, and lifetime offspring. To identify, whether or not exposure to litter leachates provokes an internal oxidative stress in the exposed animals we measured the superoxide anion radical scavenging capacity via photoluminescence. Except of P. abies, exposure to the leachates reduced this antioxidant capacity by approximately 50%. Leachate exposures, except that of Quercus, increased body size and extended lifespan; furthermore, particularly the leachates of both Picea species significantly increased the offspring numbers. This unexpected behavior of exposed Moina may be based on food supplements (e.g., high carbohydrate contents) in the leachates or on yet to be identified regulatory pathways of energy allocation. Overall, our results suggest that the potentially adverse effects of litter leachates can be overruled by either bacterial-growth supporting fractions in the leachates or an internal compensation mechanism in the Moina individuals.

Cyanobacterial Xenobiotics as Evaluated by a Caenorhabditis elegans Neurotoxicity Screening Test

In fresh waters cyanobacterial blooms can produce a variety of toxins, such as microcystin variants (MCs) and anatoxin-a (ANA). ANA is a well-known neurotoxin, whereas MCs are hepatotoxic and, to a lesser degree, also neurotoxic. Neurotoxicity applies especially to invertebrates lacking livers. Current standardized neurotoxicity screening methods use rats or mice. However, in order to minimize vertebrate animal experiments as well as experimental time and effort, many investigators have proposed the nematode Caenorhabditis elegans as an appropriate invertebrate model. Therefore, four known neurotoxic compounds (positive compounds: chlorpyrifos, abamectin, atropine, and acrylamide) were chosen to verify the expected impacts on autonomic (locomotion, feeding, defecation) and sensory (thermal, chemical, and mechanical sensory perception) functions in C. elegans. This study is another step towards successfully establishing C. elegans as an alternative neurotoxicity model. By using this protocol, anatoxin-a adversely affected locomotive behavior and pharyngeal pumping frequency and, most strongly, chemotactic and thermotactic behavior, whereas MC-LR impacted locomotion, pumping, and mechanical behavior, but not chemical sensory behavior. Environmental samples can also be screened in this simple and fast way for neurotoxic characteristics. The filtrate of a Microcystis aeruginosa culture, known for its hepatotoxicity, also displayed mild neurotoxicity (modulated short-term thermotaxis). These results show the suitability of this assay for environmental cyanotoxin-containing samples.