Combating Age-Associated Immune Decline Using Kynurenine Pathway Interventions

Select kynurenine pathway interventions extend lifespan in invertebrate models and are of interest in treating age-associated diseases. Kynurenine pathway activity is responsive to inflammatory signaling, and we are evaluating the potential for these interventions to increase pathogen resistance and curtail age-associated immune decline in Caenorhabditis elegans and mammals. The kynurenine pathway facilitates the catabolism of tryptophan to nicotinamide adenine dinucleotide (NAD). Our lab has found that supplementing the kynurenine metabolite 3-hydroxyanthranilic acid (3HAA) or inhibiting the enzyme 3HAA dioxygenase (HAAO) extends lifespan in C. elegans. 3HAA has demonstrated pro/anti-inflammatory properties in mammals, suggesting a potential role in immune function. C. elegans have a primitive immune system that lacks an adaptive element, but it recapitulates aspects of innate immune signaling and pathogen response. I hypothesize kynurenine pathway interventions that impact C. elegans’ lifespan similarly improve pathogen resistance and immunity. Interventions within the kynurenine pathway are capable of differentially impacting pathogenesis and lifespan of C. elegans challenged with Psuedomonas aeruginosa. C. elegans subjected to select lifespan-extending kynurenine pathway interventions fared better when challenged with P. aeruginosa at older ages. Additionally, fluorescent infection tracking has displayed decreased infection rates in worms with elevated 3HAA. Our data suggests pro-immune activity is facilitated by 3HAA acting downstream of the dbl-1 pathway in addition to directly inhibiting bacterial growth. Our goal is to discover the mechanism(s) through which the kynurenine pathway interacts with immune function in animals and identify potential targets for clinical therapy in aging populations.

Mammalian and Invertebrate Models as Complementary Tools for Gaining Mechanistic Insight on Muscle Responses to Spaceflight

Bioinformatics approaches have proven useful in understanding biological responses to spaceflight. Spaceflight experiments remain resource intensive and rare. One outstanding issue is how to maximize scientific output from a limited number of omics datasets from traditional animal models including nematodes, fruitfly, and rodents. The utility of omics data from invertebrate models in anticipating mammalian responses to spaceflight has not been fully explored. Hence, we performed comparative analyses of transcriptomes of soleus and extensor digitorum longus (EDL) in mice that underwent 37 days of spaceflight. Results indicate shared stress responses and altered circadian rhythm. EDL showed more robust growth signals and Pde2a downregulation, possibly underlying its resistance to atrophy versus soleus. Spaceflight and hindlimb unloading mice shared differential regulation of proliferation, circadian, and neuronal signaling. Shared gene regulation in muscles of humans on bedrest and space flown rodents suggest targets for mitigating muscle atrophy in space and on Earth. Spaceflight responses of C. elegans were more similar to EDL. Discrete life stages of D. melanogaster have distinct utility in anticipating EDL and soleus responses. In summary, spaceflight leads to shared and discrete molecular responses between muscle types and invertebrate models may augment mechanistic knowledge gained from rodent spaceflight and ground-based studies.

The Tentacular Spectacular: Evolution of Regeneration in Sea Anemones

Sea anemones vary immensely in life history strategies, environmental niches and their ability to regenerate. While the sea anemone Nematostella vectensis is the starlet of many key regeneration studies, recent work is emerging on the diverse regeneration strategies employed by other sea anemones. This manuscript will explore current molecular mechanisms of regeneration employed by non-model sea anemones Exaiptasia diaphana (an emerging model species for coral symbiosis studies) and Calliactis polypus (a less well-studied species) and examine how these species compare to the model sea anemone N. vectensis. We summarize the field of regeneration within sea anemones, within the greater context of phylum Cnidaria and in other invertebrate models of regeneration. We also address the current knowledge on two key systems that may be implemented in regeneration: the innate immune system and developmental pathways, including future aspects of work and current limitations.

Galleria mellonella as a novel eco-friendly in vivo approach for the assessment of the toxicity of medicinal plants

The evaluation of medicinal plants toxicity is a prerequisite prior their usage. The vertebrate models used for this purpose are often the object of ethical consideration. Though invertebrate models including Galleria mellonella have shown their ability to be used to assess various products toxicity, to our knowledge, G. mellonella has never been exploited to determine the toxicity of medicinal plants. In this study, the toxicity of hydroalcoholic and aqueous extracts of seven (7) Cameroonian medicinal plants namely leaves of Cymbopogon citratus (DC.) Stapf, Moringa oleifera Lam and Vernonia amygdalina Delile; barks of Cinchona officinalis and Enantia chloranta Oliv; barks and seeds of Garcinia lucida Vesque and leaves and seeds of Azadirachta indica (Neem) were evaluated using the larval form of the Greater Wax Moth (Galleria mellonella). The median lethal doses (LD50), 90% lethal doses (LD90) and 100% lethal doses were successfully determined using the spline cubic survival curves and equations from the data obtained on the survival rate of G. mellonella 24h after the injection with the extracts. The LD50 values varied from 3.90 g/kg bw to >166.67 g/kg bw and the pattern of toxicity observed was in accordance with previous investigations on the plant materials concerned. The results obtained in this study suggest that G. mellonella can be used as a sensitive, reliable, and robust eco-friendly model to gauge the toxicity of medicinal plants. Thus, avoid the sacrifice of vertebrate models often used for this purpose to limit ethical concerns.

Oxytocin Involvement in Body Composition Unveils the True Identity of Oxytocin

The origin of the Oxytocin/Vasopressin system dates back about 600 million years. Oxytocin (Oxt) together with Vasopressin (VP) regulate a diversity of physiological functions that are important for osmoregulation, reproduction, metabolism, and social behavior. Oxt/VP-like peptides have been identified in several invertebrate species and they are functionally related across the entire animal kingdom. Functional conservation enables future exploitation of invertebrate models to study Oxt’s functions not related to pregnancy and the basic mechanisms of central Oxt/VP signaling. Specifically, Oxt is well known for its effects on uteri contractility and milk ejection as well as on metabolism and energy homeostasis. Moreover, the striking evidence that Oxt is linked to energy regulation is that Oxt- and Oxytocin receptor (Oxtr)-deficient mice show late onset obesity. Interestingly Oxt−/− or Oxtr−/− mice develop weight gain without increasing food intake, suggesting that a lack of Oxt reduce metabolic rate. Oxt is expressed in a diversity of skeletal muscle phenotypes and regulates thermogenesis and bone mass. Oxt may increases skeletal muscle tonicity and/or increases body temperature. In this review, the author compared the three most recent theories on the effects of Oxt on body composition.

The Genome of the Softshell Clam Mya arenaria and the Evolution of Apoptosis

Apoptosis is a fundamental feature of multicellular animals and is best understood in mammals, flies, and nematodes, with the invertebrate models being thought to represent a condition of ancestral simplicity. However, the existence of a leukemia-like cancer in the softshell clam Mya arenaria provides an opportunity to re-evaluate the evolution of the genetic machinery of apoptosis. Here, we report the whole-genome sequence for M. arenaria which we leverage with existing data to test evolutionary hypotheses on the origins of apoptosis in animals. We show that the ancestral bilaterian p53 locus, a master regulator of apoptosis, possessed a complex domain structure, in contrast to that of extant ecdysozoan p53s. Further, ecdysozoan taxa, but not chordates or lophotrochozoans like M. arenaria, show a widespread reduction in apoptosis gene copy number. Finally, phylogenetic exploration of apoptosis gene copy number reveals a striking linkage with p53 domain complexity across species. Our results challenge the current understanding of the evolution of apoptosis and highlight the ancestral complexity of the bilaterian apoptotic tool kit and its subsequent dismantlement during the ecdysozoan radiation.

Diversification of Leech Proboscis Structure According to Prey Ingestion Behavior

Background Adaptive radiation is a phenomenon in which various organs, depending on their diet and circumstance, are diversified morphologically or functionally as animals adapt to the environment. Although previous studies on changes caused by various external pressures have been well studied, the evidence for variation in invertebrates is not well known. We used freshwater leeches as an invertebrate model to observe their specific trophic niche and diversity of ingestion organ. Our results show convergent evolution according to structural changes through a representative species Alboglossiphonia sp., and the origin from common ancestor due to the remaining fluid ingestion behavior of the larval stages as a vestige.Results We identified the feeding behavior of rhynchobdellid leeches, which have the proboscis. Alboglossiphonia sp. swallows the entire prey using its proboscis, whereas proboscis leeches exhibit typical fluid-sucking behavior. We observed that proboscis of fluid-sucking leeches encompasses compartmentalized and dense muscle layers. In contrast, macrophagous leeches have relatively simple esophagus structures. To address whether the different feeding behaviors were intrinsic, we investigated the behavioral pattern and muscle arrangement in the earlier developmental stage of rhynchobdellid leeches. Interestingly, juveniles of the macrophagous leech as well as fluid-sucking leeches have the proboscis with the compartmentalized muscle layers and exhibit fluid-sucking behaviorsConclusions Animals have adapted various ways to obtain the energy needed for their survival. Diversification and evolution of ingestion methods across species further exhibit the functional morphology of the ingestion organ. However, information on ingestion behavior and internal structure is still lack and unclear, especially in invertebrate models. Our results suggest that the proboscis leeches have originated from the common fluid-sucking glossiphoniid ancestor and species diversification has led to modifications in the structure of the feeding tube. Together, leeches represent a comparative model for esophagus development according to the ingestion pattern based on diverse muscular arrangement in proboscis.

The design and implementation of restraint devices for the injection of pathogenic microorganisms into Galleria mellonella

The injection of laboratory animals with pathogenic microorganisms poses a significant safety risk because of the potential for injury by accidental needlestick. This is especially true for researchers using invertebrate models of disease due to the small size of the animals and the required precision and accuracy of the injection. The immobilization of the greater wax moth larvae (Galleria mellonella) is often achieved by grasping a larva firmly between finger and thumb. Needle resistant gloves or forceps can be used to reduce the risk of a needlestick but can result in animal injury, a loss of throughput, and inconsistencies in experimental data. Immobilization devices are commonly used for the manipulation of small mammals, and in this manuscript, we describe the construction of injection chambers that can be used to entrap and restrain G. mellonella larvae prior to injection with pathogenic microbes. These devices significantly reduce the manual handling of larvae and provide an engineering control to protection against accidental needlestick injury, while maintaining a high rate of injection.