Search

Molecular vibrations and quantum tunneling may link ligand binding to the function of pharmacological receptors. The well‐established lock‐and‐key model explains a ligand's binding and recognition by a receptor; however, a general mechanism by which receptors translate binding into activation, inactivation, or modulation remains elusive. The Vibration Theory of Olfaction was proposed in the 1930s to explain this subset of receptor‐mediated phenomena by correlating odorant molecular vibrations to smell, but a mechanism was lacking. In the 1990s, inelastic electron tunneling was proposed as a plausible mechanism for translating molecular vibration to odorant physiology. More recently, studies of ligands' vibrational spectra and the use of deuterated ligand analogs have provided helpful information to study this admittedly controversial hypothesis in metabotropic receptors other than olfactory receptors. In the present work, based in part on published experiments from our laboratory using planarians as an experimental organism, I will present a rationale and possible experimental approach for extending this idea to ligand‐gated ion channels. [ABSTRACT FROM AUTHOR]

This study explored the potential of Andrographis paniculata in regulating redox imbalance using planaria as a model organism. Decapitated planaria were regenerated in water, A. paniculata extract, hydrogen peroxide, and a combination of hydrogen peroxide and A. paniculata. The growth was 26% higher, locomotion 15% higher, and protein 35% higher in A. paniculata group. The stressed group had 33% higher growth, 17% higher locomotion, and 36% higher protein when treated with A. paniculata. In conclusion, A. paniculata accelerated the regeneration and minimized the stress induced by hydrogen peroxide and hence can be a promising option for the prevention of neurodegenerative diseases. [ABSTRACT FROM AUTHOR]

Wildlife is increasingly exposed to sublethal transient cancer risk factors, including mutagenic substances, which activates their anti-cancer defences, promotes tumourigenesis, and may negatively impact populations. Little is known about how exposure to cancer risk factors impacts the behaviour of wildlife. Here, we investigated the effects of a sublethal, short-term exposure to a carcinogen at environmentally relevant concentrations on the activity patterns of wild Girardia tigrina planaria during a two-phase experiment, consisting of a 7-day exposure to cadmium period followed by a 7-day recovery period. To comprehensively explore the effects of the exposure on activity patterns, we employed the double hierarchical generalized linear model framework which explicitly models residual intraindividual variability in addition to the mean and variance of the population. We found that exposed planaria were less active compared to unexposed individuals and were able to recover to pre-exposure activity levels albeit with a reduced variance in activity at the start of the recovery phase. Planaria showing high activity levels were less predictable with larger daily activity variations and higher residual variance. Thus, the shift in behavioural variability induced by an exposure to a cancer risk factor can be quantified using advanced tools from the field of behavioural ecology. This is required to understand how tumourous processes affect the ecology of species. [ABSTRACT FROM AUTHOR]

Cover Photograph: Whole‐mount immunostaining of DjPiwiA (magenda) of an X ray‐irradiated planaria injected with isolated neoblasts (see Ishida pp 577–590). [ABSTRACT FROM AUTHOR]

Advanced oxidative processes (AOPs) are procedures used for the treatment of wastewater based on the generation of free radicals, such as hydroxyl (•OH) and carbonate anion (CO3•−) radicals. However, although contaminants are degraded in these processes, the by-products generated in this transformation can be a greater source of toxicity than the original compound, making ecotoxicological tests essential for monitoring the efficacy of these treatment processes. In this study, we examined the ecotoxicity of AOP by-products generated using peracetic acid (PAA) and sodium carbonate, with and without solar radiation, for the degradation of methylene blue dye, using the planarian Girardia tigrina as a test organism. Ecotoxicological tests evaluated the acute toxicity of the generated by-product in terms of lethal concentrations (LC50). Although in both assays the degradation of the dye was greater than 99%, higher toxicity was observed in the assay using PAA and carbonate in the absence of radiation. From the results obtained, we conclude that the by-product generated from the degradation of methylene blue dye by peracetic acid and sodium carbonate, with and without solar radiation, can pose risks to aquatic ecosystems if released directly into water bodies. [ABSTRACT FROM AUTHOR]

Single-cell transcriptomics has revolutionised biology allowing the quantification of gene expression in individual cells. Since each single cell contains cell type specific mRNAs, these techniques enable the classification of cell identities. Therefore, single cell methods have been used to explore the repertoire of cell types (the single cell atlas) of different organisms, including freshwater planarians. Nowadays, planarians are one of the most prominent animal models in single cell biology. They have been studied at the single cell level for over a decade using most of the available single cell methodological approaches. These include plate-based methods, such as qPCR, nanodroplet methods and in situ barcoding methods. Because of these studies, we now have a very good picture of planarian cell types and their differentiation trajectories. Planarian regenerative properties and other characteristics, such as their developmental plasticity and their capacity to reproduce asexually, ensure that another decade of single cell biology in planarians is yet to come. Here, we review these characteristics, the new biological insights that have been obtained by single-cell transcriptomics and outline the perspectives for the future. [ABSTRACT FROM AUTHOR]

Asian vegetables are important specialty crops in Florida, but no information is available on their host status to plant-parasitic nematodes, in particular to Meloidogyne spp. Meloidogyne spp. are one of the main soilborne pathogens in Florida and cause considerable damage to many vegetables. We conducted a nematode survey on 10 Asian vegetable farms in Hillsborough County, FL, all owned and operated by Vietnamese growers. A total of 66 root samples representing 20 commonly grown Asian vegetables were collected and evaluated for the presence of Meloidogyne spp. Meloidogyne females were dissected from roots and collected for molecular identification using primer sets TRNAH/MRH106 and MORF/MTHIS, followed by restricting PCR products using HinfI and MnlI restriction enzymes. Species-specific primers were used to confirm Meloidogyne spp. and mtDNA sequencing was used when needed to confirm the identification. We identified four Meloidogyne spp., M. arenaria, M. enterolobii, M. haplanaria, and M. incognita, infecting Asian vegetables. Meloidogyne enterolobii was the most commonly found Meloidogyne spp. and accounted for almost 75% of positive samples. Our study revealed that Meloidogyne spp. are widespread in Asian vegetable farms and especially M. enterolobii. Meloidogyne enterolobii is considered one of the most damaging Meloidogyne spp., and its high prevalence in Asian vegetable farms is of concern. Asian vegetable growers need to be made aware of this nematode as well of the threat of other Meloidogyne spp. to production. Nematode management programs need to be implemented in these Asian vegetable farms to help growers overcome the loss caused by Meloidogyne spp. and ensure the sustainability of their farms. [ABSTRACT FROM AUTHOR]

Exposure to high levels of ionizing γ radiation leads to irreversible DNA damage and cell death. Here, we establish that exogenous application of electric stimulation enables cellular plasticity and the re-establishment of stem cell activity in tissues damaged by ionizing radiation. We show that subthreshold direct current stimulation (DCS) rapidly restores pluripotent stem cell populations previously eliminated by lethally γ-irradiated tissues of the planarian flatworm Schmidtea mediterranea. Our findings reveal that DCS enhances DNA repair, transcriptional activity, and cell cycle entry in post-mitotic cells. These responses involve rapid increases in cytosolic Ca2+ concentration through the activation of L-type Cav channels and intracellular Ca2+ stores, leading to the activation of immediate early genes and ectopic expression of stem cell markers in post-mitotic cells. Overall, we show the potential of electric current stimulation to reverse the damaging effects of high-dose γ radiation in adult tissues. Furthermore, our results provide mechanistic insights describing how electric stimulation effectively translates into molecular responses capable of regulating fundamental cellular functions without the need for genetic or pharmacological intervention. [ABSTRACT FROM AUTHOR]

One approach to elucidating the principles of regeneration is to investigate mechanisms that regenerate a target organ. Planarian eyes are discrete, visible structures that are dispensable for viability, making them powerful for studying the logic of regeneration. Fate specification in eye regeneration occurs in stem cells (neoblasts), generating eye progenitors. Eye progenitor production is not responsive to the presence or absence of the eye, with regeneration explained by constant progenitor production in the appropriate positional environment. Eye progenitors display coarse spatial specification. A combination of eye-extrinsic cues and self-organization with differentiated eye cells dictate where migratory eye progenitors target. Finally, guidepost-like cells influence regenerating axons to facilitate the restoration of eye circuitry. These findings from the eye as a case study present a model that explains how regeneration can occur. [ABSTRACT FROM AUTHOR]

Simultaneous longitudinal imaging across multiple conditions and replicates has been crucial for scientific studies aiming to understand biological processes and disease. Yet, imaging systems capable of accomplishing these tasks are economically unattainable for most academic and teaching laboratories around the world. Here, we propose the Picroscope, which is the first low-cost system for simultaneous longitudinal biological imaging made primarily using off-the-shelf and 3D-printed materials. The Picroscope is compatible with standard 24-well cell culture plates and captures 3D z-stack image data. The Picroscope can be controlled remotely, allowing for automatic imaging with minimal intervention from the investigator. Here, we use this system in a range of applications. We gathered longitudinal whole organism image data for frogs, zebrafish, and planaria worms. We also gathered image data inside an incubator to observe 2D monolayers and 3D mammalian tissue culture models. Using this tool, we can measure the behavior of entire organisms or individual cells over long-time periods. Ly et al. report a multi-well imaging system (the Picroscope) that can perform longitudinal bright-field z-stack imaging for 24 wells simultaneously. They demonstrate its capability by imaging zebrafish embryos, planaria worms, development of Xenopus tropicalis over a 28-h period and embryonic stem cell monolayer and organoids inside an incubator. [ABSTRACT FROM AUTHOR]

In developmental biology, the regulation of stem cell plasticity and differentiation remains an open question. CBP(CREB-binding protein)/p300 is a conserved gene family that functions as a transcriptional co-activator and plays important roles in a wide range of cellular processes, including cell death, the DNA damage response, and tumorigenesis. The acetyl transferase activity of CBPs is particularly important, as histone and non-histone acetylation results in changes in chromatin architecture and protein activity that affect gene expression. Many studies have described the conserved functions of CBP/p300 in stem cell proliferation and differentiation. The planarian Schmidtea mediterranea is an excellent model for the in vivo study of the molecular mechanisms underlying stem cell differentiation during regeneration. However, how this process is regulated genetically and epigenetically is not well-understood yet. We identified 5 distinct Smed-cbp genes in S. mediterranea that show different expression patterns. Functional analyses revealed that Smed-cbp-2 appears to be essential for stem cell maintenance. On the other hand, the silencing of Smed-cbp-3 resulted in the growth of blastemas that were apparently normal, but remained largely unpigmented and undifferentiated. Smed-cbp-3 silencing also affected the differentiation of several cell lineages including neural, epidermal, digestive, and excretory cell types. Finally, we analysed the predicted interactomes of CBP-2 and CBP-3 as an initial step to better understand their functions in planarian stem cell biology. Our results indicate that planarian cbp genes play key roles in stem cell maintenance and differentiation. [Display omitted] • The CREB-binding protein family is expanded in planarians. • The silencing of cbp-2 and cbp-3 result in rather complementary phenotypes. • cbp-2 is required for planarian survival and stem cell maintenance. • cbp-3 is required for the proper differentiation of multiple lineages. [ABSTRACT FROM AUTHOR]

Many biological organisms regenerate structure and function after damage. Despite the long history of research on molecular mechanisms, many questions remain about algorithms by which cells can cooperate towards the same invariant morphogenetic outcomes. Therefore, conceptual frameworks are needed not only for motivating hypotheses for advancing the understanding of regeneration processes in living organisms, but also for regenerative medicine and synthetic biology. Inspired by planarian regeneration, this study offers a novel generic conceptual framework that hypothesizes mechanisms and algorithms by which cell collectives may internally represent an anatomical target morphology towards which they build after damage. Further, the framework contributes a novel nature-inspired computing method for self-repair in engineering and robotics. Our framework, based on past in vivo and in silico studies on planaria, hypothesizes efficient novel mechanisms and algorithms to achieve complete and accurate regeneration of a simple in silico flatwormlike organism from any damage, much like the body-wide immortality of planaria, with minimal information and algorithmic complexity. This framework that extends our previous circular tissue repair model integrates two levels of organization: tissue and organism. In Level 1, three individual in silico tissues (head, body, and tail—each with a large number of tissue cells and a single stem cell at the centre) repair themselves through efficient local communications. Here, the contribution extends our circular tissue model to other shapes and invests them with tissue-wide immortality through an information field holding the minimum body plan. In Level 2, individual tissues combine to form a simple organism. Specifically, the three stem cells form a network that coordinates organism-wide regeneration with the help of Level 1. Here we contribute novel concepts for collective decision-making by stem cells for stem cell regeneration and large-scale recovery. Both levels (tissue cells and stem cells) represent networks that perform simple neural computations and form a feedback control system. With simple and limited cellular computations, our framework minimises computation and algorithmic complexity to achieve complete recovery. We report results from computer simulations of the framework to demonstrate its robustness in recovering the organism after any injury. This comprehensive hypothetical framework that significantly extends the existing biological regeneration models offers a new way to conceptualise the information-processing aspects of regeneration, which may also help design living and non-living self-repairing agents. [ABSTRACT FROM AUTHOR]

Nervous systems' computational abilities are an evolutionary innovation, specializing and speed-optimizing ancient biophysical dynamics. Bioelectric signalling originated in cells' communication with the outside world and with each other, enabling cooperation towards adaptive construction and repair of multicellular bodies. Here, we review the emerging field of developmental bioelectricity, which links the field of basal cognition to state-of-the-art questions in regenerative medicine, synthetic bioengineering and even artificial intelligence. One of the predictions of this view is that regeneration and regulative development can restore correct large-scale anatomies from diverse starting states because, like the brain, they exploit bioelectric encoding of distributed goal states--in this case, pattern memories. We propose a new interpretation of recent stochastic regenerative phenotypes in planaria, by appealing to computational models of memory representation and processing in the brain. Moreover, we discuss novel findings showing that bioelectric changes induced in planaria can be stored in tissue for over a week, thus revealing that somatic bioelectric circuits in vivo can implement a long-term, re-writable memory medium. A consideration of the mechanisms, evolution and functionality of basal cognition makes novel predictions and provides an integrative perspective on the evolution, physiology and biomedicine of information processing in vivo. This article is part of the theme issue 'Basal cognition: multicellularity, neurons and the cognitive lens'. [ABSTRACT FROM AUTHOR]

The recently elucidated proresolving conjugates in tissue regeneration (CTR) maresin-CTR (MCTR), protectin-CTR (PCTR), and resolvin-CTR (RCTR), termed cysteinyl-specialized proresolving mediators (cys-SPMs) each promotes regeneration, controls infection, and accelerates resolution of inflammation. Here, we sought evidence for cys-SPM activation of primordial pathways in planaria (Dugesia japonica) regeneration that might link resolution of inflammation and regeneration. On surgical resection, planaria regeneration was enhanced with MCTR3, PCTR3, or RCTR3 (10 nM), each used for RNA sequencing. The three cys-SPMs shared up-regulation of 175 known transcripts with fold-change > 1.25 and combined false discovery rate (FDR) < 0.002, and 199 canonical pathways (FDR < 0.25), including NF-κB pathways and an ortholog of human TRAF3 (TNFR-associated factor 3). Three separate pathway analyses converged on TRAF3 upregulation by cys-SPMs. With human macrophages, three cys-SPMs each dose-dependently increased TRAF3 expression in a cAMPPKA-dependent manner. TRAF3 overexpression in macrophages enhanced Interleukin-10 (IL-10) and phagocytosis of Escherichia coli. IL-10 also increased phagocytosis in a dose-dependent manner. Silencing of mouse TRAF3 in vivo significantly reduced IL-10 and macrophage phagocytosis. TRAF3 silencing in vivo also relieved cys-SPMs' actions in limiting polymorphonuclear neutrophil in E. coli exudates. These results identify cys-SPM-regulated pathways in planaria regeneration, uncovering a role for TRAF3/IL-10 in regulating mammalian phagocyte functions in resolution. Cys-SPM activation of TRAF3 signaling is a molecular component of both regeneration and resolution of infectious inflammation. [ABSTRACT FROM AUTHOR]

Metazoan body plans combine well-defined primary, secondary, and in many bilaterians, tertiary body axes with structural asymmetries at multiple scales. Despite decades of study, how axis-defining symmetries and system-defining asymmetries co-emerge during both evolution and development remain open questions. Regeneration studies in asexual planaria have demonstrated an array of viable forms with symmetrized and, in some cases, duplicated body axes. We suggest that such forms may point toward an ancestral eumetazoan form with characteristics of both cnidarians and placazoa. [ABSTRACT FROM AUTHOR]

The coordination of tissue-level polarity with organism-level polarity is crucial in development, disease, and regeneration. Here, we characterize a new example of large-scale control of dynamic remodeling of body polarity. Exploiting the flexibility of the body plan in regenerating planarians, we used mirror duplication of the primary axis to show how established tissue-level polarity adapts to new organism-level polarity. Characterization of epithelial planar cell polarity revealed a remarkable reorientation of tissue polarity in double-headed planarians. This reorientation of cilia occurs even following irradiation-induced loss of all stem cells, suggesting independence of the polarity change from the formation of new cells. The presence of the two heads plays an important role in regulating the rate of change in overall polarity. We further present data that suggest that the nervous system itself adapts its polarity to match the new organismal anatomy as revealed by changes in nerve transport driving distinct regenerative outcomes. Thus, in planaria tissue-level polarity can dynamically reorient to match the organism-level anatomical configuration. • Cilia-driven flow in planaria adapts to new body morphology. • Reorientation of flow occurs over long timeframe. • Presence of heads in double-headed planaria drives cilia reorientation. • Nervous system polarity adapts to induction of double-headed morphology. [ABSTRACT FROM AUTHOR]

There are benefits to both laboratory exercises and scientific modeling, and connecting the two may allow for deeper understanding and interest. Laboratory exercises provide students with opportunities to experience phenomena, but without scientific modeling, students may still lack understanding of the mechanisms at play. This article describes an example of how a traditional laboratory exercise on planarian regeneration is enhanced with a modeling activity on cell signaling. [ABSTRACT FROM AUTHOR]

Planarians are widely used animal models for studies in regeneration, developmental biology, neurobiology, and behavior. However, surprisingly little is known about other aspects of their basic biology, even though such information might help validate these flatworms as a general animal model. We hypothesized that planaria, although dependent on simple diffusion of O2 across the integument for O2 uptake, would nonetheless show changes in oxygen consumption (...O2) associated with reproductive mode (sexual or asexual), feeding (specific dynamic action; SDA), temperature (Q10 values), and photoperiod typical of those responses of more complex invertebrates. In the current experiments, routine ...O2 was measured over the range of 13-28°C in Schmidtea mediterranea and Girardia dorotocephala. At the long-term maintenance temperature of 18°C, routine ...O2 was ~13 µL O2·g-1·h-1 in the two asexual strains, but approximately twice as high (27 µL O2·g-1·h-1) in the sexual strain of S. mediterranea, suggesting a metabolic cost for sexual reproduction. Metabolic temperature sensitivity, measured by Q10, was about one to three for all three groups. All three groups showed a large (~2- to 3-fold) increase in ...O2 within a day following feeding, suggesting a large SDA effect. Starvation, causing "degrowth" in some planaria, resulted in a loss of one-third of body mass in sexual S. mediterranea but no body mass loss in either asexual strains. Collectively, these data indicate that, while being a relatively simple flatworm with no dedicated respiratory or circulatory system, their metabolic physiological responses are quite similar to those shown by more complex invertebrates and vertebrates, contributing to their validation as an animal model. [ABSTRACT FROM AUTHOR]

Planarians are widely used animal models for studies in regeneration, developmental biology, neurobiology, and behavior. However, surprisingly little is known about other aspects of their basic biology, even though such information might help validate these flatworms as a general animal model. We hypothesized that planaria, although dependent on simple diffusion of O2 across the integument for O2 uptake, would nonetheless show changes in oxygen consumption (V O2) associated with reproductive mode (sexual or asexual), feeding (specific dynamic action; SDA), temperature (Q10 values), and photoperiod typical of those responses of more complex invertebrates. In the current experiments, routine V O2 was measured over the range of 13-28°C in Schmidtea mediterranea and Girardia dorotocephala. At the long-term maintenance temperature of 18°C, routineV O2 was ~13 μL O2·g-1·h-1 in the two asexual strains, but approximately twice as high (27 μL O2·g-1·h-1) in the sexual strain of S. mediterranea, suggesting a metabolic cost for sexual reproduction. Metabolic temperature sensitivity, measured by Q10, was about one to three for all three groups. All three groups showed a large (~2- to 3-fold) increase inV O2 within a day following feeding, suggesting a large SDA effect. Starvation, causing "degrowth" in some planaria, resulted in a loss of one-third of body mass in sexual S. mediterranea but no body mass loss in either asexual strains. Collectively, these data indicate that, while being a relatively simple flatworm with no dedicated respiratory or circulatory system, their metabolic physiological responses are quite similar to those shown by more complex invertebrates and vertebrates, contributing to their validation as an animal model. [ABSTRACT FROM AUTHOR]

Motile cilia propel directed cell movements and sweep fluids across the surface of tissues. Orthologs of Dynein Assembly Factor with WD Repeat Domains 1 (DAW1) support normal ciliary beating by enhancing delivery of dynein complexes to axonemal microtubules. DAW1 mutations in vertebrates result in multiple developmental abnormalities and early or prenatal lethality, complicating functional assessment of DAW1 in adult structures. Planarian flatworms maintain cellular homeostasis and regenerate through differentiation of adult pluripotent stem cells, and systemic RNA‐interference (RNAi) can be induced to analyze gene function at any point after birth. A single ortholog of DAW1 was identified in the genome of the planarian Schmidtea mediterranea (Smed‐daw1). Smed‐DAW1 is composed of eight WD repeats, which are 55% identical to the founding member of this protein family (Chlamydomonas reinhardtii ODA16) and 58% identical to human DAW1. Smed‐daw1 is expressed in the planarian epidermis, protonephridial excretory system, and testes, all of which contain cells functionally dependent on motile cilia. Smed‐daw1 RNAi resulted in locomotion defects and edema, which are phenotypes characteristic of multiciliated epidermis and protonephridial dysfunction, respectively. Changes in abundance or length of motile cilia were not observed at the onset of phenotypic manifestations upon Smed‐daw1 RNAi, corroborating with studies showing that DAW‐1 loss of function leads to aberrant movement of motile cilia in other organisms, rather than loss of cilia per se. However, extended RNAi treatments did result in shorter epidermal cilia and decreased abundance of ciliated protonephridia, suggesting that Smed‐daw1 is required for homeostatic maintenance of these structures in flatworms. [ABSTRACT FROM AUTHOR]

Planaria is an ideal system to study factors involved in regeneration and tissue homeostasis. Little is known about the role of metabolites and small molecules in stem cell maintenance and lineage specification in planarians. Using liquid chromatography and mass spectrometry (LC‐MS)‐based quantitative metabolomics, we determined the relative levels of metabolites in stem cells, progenitors, and differentiated cells of the planarian Schmidtea mediterranea. Tryptophan and its metabolic product serotonin are significantly enriched in stem cells and progenitor population. Serotonin biosynthesis in these cells is brought about by a noncanonical enzyme, phenylalanine hydroxylase. Knockdown of Smed‐pah leads to complete disappearance of eyes in regenerating planaria, while exogenous supply of serotonin and its precursor rescues the eyeless phenotype. Our results demonstrate a key role for serotonin in eye regeneration. [ABSTRACT FROM AUTHOR]

Flatworms can very rapidly attach to and detach from many substrates. In the presented work, we analysed the adhesive system of the marine proseriate flatworm Minona ileanae. We used light-, scanning- and transmission electron microscopy to analyse the morphology of the adhesive organs, which are located at the ventral side of the tail-plate. We performed transcriptome sequencing and differential RNA-seq for the identification of tail-specific transcripts. Using in situ hybridization expression screening, we identified nine transcripts that were expressed in the cells of the adhesive organs. Knock-down of five of these transcripts by RNA interference led to a reduction of the animal's attachment capacity. Adhesive proteins in footprints were confirmed using mass spectrometry and antibody staining. Additionally, lectin labelling of footprints revealed the presence of several sugar moieties. Furthermore, we determined a genome size of about 560 Mb for M. ileanae. We demonstrated the potential of Oxford Nanopore sequencing of genomic DNA as a cost-effective tool for identifying the number of repeats within an adhesive protein and for combining transcripts that were fragments of larger genes. A better understanding of the molecules involved in flatworm bioadhesion can pave the way towards developing innovative glues with reversible adhesive properties. This article is part of the theme issue 'Transdisciplinary approaches to the study of adhesion and adhesives in biological systems'. [ABSTRACT FROM AUTHOR]

Intrinsic reaction coordinate (IRC) torsional potentials were calculated for N2O4 and N2O3 based on optimized B3LYP/aug-cc-pVDZ geometries of the respective 90°-twisted saddle points. These potentials were refined by obtaining CCSD(T)/aug-cc-pVXZ energies [in the complete basis set (CBS) limit] of points along the IRC. A comparison is made between these ab initio potentials and an analytical form based on a two-term cosine expansion in terms of the N–N dihedral angle. The shapes of these two potential curves are in close agreement. The torsional barriers in N2O4 and N2O3 obtained from the CCSD(T)/CBS//B3LYP/aug-cc-pVDZ calculations are 2333 and 1704 cm-1, respectively. For N2O4 the torsion fundamental frequency from the IRC potential is 87.06 cm-1, which is in good agreement with the experimentally reported value of 81.73 cm-1. However, in the case of N2O3 the torsional frequency found from the IRC potential, 144 cm-1, is considerably larger than the reported experimental values 63–76 cm-1. Consistent with this discrepancy, the torsional barrier obtained from several different calculations, 1417–1718 cm-1, is higher than the value of 350 cm-1 deduced from experimental studies. It is suggested that the assignment of the torsional mode in N2O3 should be reexamined. N2O4 and N2O3 exhibit strong hyperconjugative interactions of in-plane O lone pairs with the central N–N σ* antibond. Hyperconjugative stabilization is somewhat stronger at the planar geometries because 1,4 interactions of lone pairs on cis O atoms promote delocalization of electrons into the N–N antibond. Calculations therefore suggest that the torsional barriers in these molecules arise principally from a combination of 1,4 interactions and hyperconjugation. [ABSTRACT FROM AUTHOR]

Photomotor response assay (PMR) is very useful in an ecotoxicological context because it allows evaluation of behavioral response to potential toxic compounds. However, a lack of procedure standardization makes results comparison difficult between labs and organisms. Here, we aimed to propose five different tools to standardize the PMR procedure so that it may be applied to all model species, regarding: (1) the minimum total sample size, (2) the acclimation period, (3) the number and duration of light and dark phases alternation, (4) the measured behavior, and (5) the statistical analysis. As an example of procedure application, we analyzed the effect of an exposure to the antibiotic gentamicin on the locomotion behavior during PMR in an invertebrate species: the asexual freshwater planaria Schmidtea mediterranea. We encourage future studies using PMR to follow these five tools to improve data analysis and results comparability. • PMR allows evaluation of behavioral response to potential toxic compounds. • Literature highlights the lack of procedure standardization between labs/organisms. • We proposed five tools to standardize PMR procedure. • Standardized PMR procedure was tested on gentamicin-exposed planaria S. mediterranea. • Standardized PMR procedure is suitable for ecotoxicological studies in invertebrates. [ABSTRACT FROM AUTHOR]

The objective of the present study was to develop a model of avoidance learning and its extinction in planarians (Schmidtea mediterranea). Based on previous experiments showing conditioned place preference, we developed a procedure to investigate conditioned place avoidance (CPA) using shock as an unconditioned stimulus (US) and an automated tracking system to record the animals' behaviour. In Experiment 1, we assessed the unconditioned properties of different shock intensities by measuring post shock activity. In two subsequent experiments we investigated CPA using different designs, surfaces as conditioned stimuli (CSs; rough and smooth), and different US intensities (5 V and 10 V). In general, we observed the successful development of CPA. However, CPA was stronger with higher shock intensities, and we found that, in our preparation, a rough surface is best at entering into an association with the shock than a smooth surface. Finally, we also observed extinction of CPA. The evidence of CPA and its extinction in flatworms validates the planaria as a pre-clinical model for the study of avoidance learning, a hallmark of anxiety disorders. • Planarians are sensitive to different shock intensities. • Planarians shocked in a distinctive context develop conditioned place avoidance. • Context non-reinforced exposure results in extinction of the avoidance response. • Planarians can be used as an invertebrate model to study avoidance behaviour. [ABSTRACT FROM AUTHOR]

Freshwater planarian are emerging as a valuable in vivo model for (eco) toxicological studies, but the lack of harmonization of procedures between laboratories remains a challenge. This study aimed to optimize, automate and select the best behavioural tests and analyse the potential of different planarian species as models for toxicological assessment. We implemented four tests: exploration, photomotor response, Tapping and Planarian Light Dark Test, on different planaria species using the DanioVision system. We conclude that the exploration assay performed in 24 well-plate at 10,000 lux is the one that is robust and reliable for toxicological studies with planaria. Dugesia japonica and Schmidtea mediterranea have proved to be sensitive models for toxicological screening of organophosphorus pesticides through behavioural analysis. Under necessary experimental conditions, the motility baseline in controls, for both species allowed the detection of behavioural changes, making both good models for behavioural testing in (eco) toxicological context. • Freshwater planarians are a promising in-vivo (eco)toxicological screening tool • Optimization of behavioural tests and selection of the best planarian species • Exploration test is robust and reliable for toxicological assessment of OPs on planaria • D.japonica and S.mediterranea are both good models for behavioural testing in toxicological context [ABSTRACT FROM AUTHOR]

• Histamine functions as a punisher in vertebrate models. • We investigated if planarians would develop conditioned place aversion to histamine. • Planarians exposed to histamine in a dark environment spent more time in the light. • Histamine did not completely overcome innate preference for the dark. Although histamine functions as a punisher in vertebrate models, its potential aversive effects in invertebrates has gone largely unexamined. We investigated if planaria would develop conditioned place aversions to histamine. In the absence of any training, planaria readily move away from a light source. However, planaria will develop conditioned place preferences for lighted areas if those areas are paired with many of the same psychoactive drugs that produce conditioned place preferences in vertebrates. We confined planaria in the darkened side of a petri dish containing histamine, and they subsequently spent more time in the light than planaria confined initially to the darkened side of a petri dish containing fresh water only. This occurred whether we tested planaria individually or in groups. Pairing histamine with the light side of the petri dish had inconsistent effects, and histamine did not affect behavior under a motility assay. Although histamine altered the planaria behavior, it did not completely overcome the planaria's innate preference for darkened areas. Results add to a growing body of research showing continuity between planaria and vertebrates in the behavioral effects of psychoactive chemicals. [ABSTRACT FROM AUTHOR]

One of the most important aspects of the scientific endeavour is the definition of specific concepts as precisely as possible. However, it is also important not to lose sight of two facts: (i) we divide the study of nature into manageable parts in order to better understand it owing to our limited cognitive capacities and (ii) definitions are inherently arbitrary and heavily influenced by cultural norms, language, the current political climate, and even personal preferences, among many other factors. As a consequence of these facts, clear-cut definitions, despite their evident importance, are oftentimes quite difficult to formulate. One of the most illustrative examples about the difficulty of articulating precise scientific definitions is trying to define the concept of a brain. Even though the current thinking about the brain is beginning to take into account a variety of organisms, a vertebrocentric bias still tends to dominate the scientific discourse about this concept. Here I will briefly explore the evolution of our 'thoughts about the brain', highlighting the difficulty of constructing a universally (or even a generally) accepted formal definition of it and using planarians as one of the earliest examples of organisms proposed to possess a 'traditional', vertebrate-style brain. I also suggest that the time is right to attempt to expand our view of what a brain is, going beyond exclusively structural and taxa-specific criteria. Thus, I propose a classification that could represent a starting point in an effort to expand our current definitions of the brain, hopefully to help initiate conversations leading to changes of perspective on how we think about this concept. This article is part of the theme issue 'Liquid brains, solid brains: How distributed cognitive architectures process information'. [ABSTRACT FROM AUTHOR]

Control of axial polarity during regeneration is a crucial open question. We developed a quantitative model of regenerating planaria, which elucidates self-assembly mechanisms of morphogen gradients required for robust body-plan control. The computational model has been developed to predict the fraction of heteromorphoses expected in a population of regenerating planaria fragments subjected to different treatments, and for fragments originating from different regions along the anterior-posterior and medio-lateral axis. This allows for a direct comparison between computational and experimental regeneration outcomes. Vector transport of morphogens was identified as a fundamental requirement to account for virtually scale-free self-assembly of the morphogen gradients observed in planarian homeostasis and regeneration. The model correctly describes altered body-plans following many known experimental manipulations, and accurately predicts outcomes of novel cutting scenarios, which we tested. We show that the vector transport field coincides with the alignment of nerve axons distributed throughout the planarian tissue, and demonstrate that the head-tail axis is controlled by the net polarity of neurons in a regenerating fragment. This model provides a comprehensive framework for mechanistically understanding fundamental aspects of body-plan regulation, and sheds new light on the role of the nervous system in directing growth and form. [ABSTRACT FROM AUTHOR]

Highlights • Planarian model as an alternative to dissect DNA damage repair during tissue renewal. • DNA repair mechanisms are evolutionarily conserved in planarians. • Genetic models of DNA damage have been developed in planarians. • Regional signals affect cellular decision in the presence of DNA Damage. • DNA damage repair plays a central role during regeneration of tissues. Abstract Faithful renewal of aging and damaged tissues is central to organismal lifespan. Stem cells (SCs) generate the cellular progeny that replenish adult tissues across the body but this task becomes increasingly compromised over time. The age related decline in SC-mediated tissue maintenance is a multifactorial event that commonly affects genome integrity. The presence of DNA damage in SCs that are under continuous demand to divide poses a great risk for age-related disorders such as cancer. However, performing analysis of SCs with genomic instability and the DNA damage response during tissue renewal present significant challenges. Here we introduce an alternative experimental system based on the planaria flatworm Schmidtea mediterranea to address at the organismal level studies intersecting SC-mediated tissue renewal in the presence of genomic instability. Planaria have abundant SCs (neoblasts) that maintain high rates of cellular turnover and a variety of molecular tools have been developed to induce DNA damage and dissect how neoblasts respond to this stressor. S. mediterranea displays high evolutionary conservation of DNA repair mechanisms and signaling pathways regulating adult SCs. We describe genetically induced-DNA damage models and highlight body-wide signals affecting cellular decisions such as survival, proliferation, and death in the presence of genomic instability. We also discuss transcriptomic changes in the DNA damage response during injury repair and propose DNA repair as key component of tissue regeneration. Additional studies using planaria will provide insights about mechanisms regulating survival and growth of cells with DNA damage during tissue renewal and regeneration. [ABSTRACT FROM AUTHOR]

Abstract A key requirement of tissue/organ regeneration is the ability to induce appropriate shape in situ. Regenerated structures need to be integrated with pre-existing ones, through the combined regulation of new tissue growth and the scaling of surrounding tissues. This requires a tightly coordinated control of individual cell functions such as proliferation and stem cell differentiation. While great strides have been made in elucidating cell growth and differentiation mechanisms, how overall shape is generated during regeneration remains unknown. This is because a significant gap remains in our understanding of how cell behaviors are coordinated at the level of tissues and organs. The highly regenerative planarian flatworm has emerged as an important model for defining and understanding regenerative shape mechanisms. This review provides an overview of the main processes known to regulate tissue and animal shape during planarian regeneration: adult stem cell regulation, the reestablishment of body axes, tissue remodeling in pre-existing structures, organ scaling and the maintenance of body proportion, and the bioelectrical regulation of animal morphology. In order for the field to move forward, it will be necessary to identify shape mutants as a means to uncover the molecular mechanisms that synchronize all these separate processes to produce the worm's final regenerative shape. This knowledge will also aid efforts to define the mechanisms that control the termination of regenerative processes. [ABSTRACT FROM AUTHOR]

Abstract Scientific fields grow by accretion of knowledge brought up by succesive generations of scientists. With the field of planarian regeneration as a general background, here I give a personal account of it from the late 1960s until the late 1990s when new research groups, namely Americans, broke into it. After a short historical summary, I report how I got into regeneration, why I choose planarians, and most especially the finding and description of the current model organism Schmidtea mediterranea. Next, I concentrate on the quantitative cellular approaches to regeneration, growth, and degrowth undertaken in our lab in Barcelona, and the long struggle to set neoblasts as a totipotent/pluripotent heterogeneous population of mitotic and non-mitotic cells for homeostatic somatic cell renewal, reproduction, and blastema formation during regeneration. Finally, the first attempts by several labs to analyze regeneration in molecular terms (RNAs, proteins, mAbs, gene detection, cloning and expression) are also covered. Along this essay, I am honoured to pay due tribute to our forebears (ancestors) from Morgan and Child to the French School researchers. In spite of being often baffled by planarian 'tricks', they struggled hard to find new techniques, new ways, and new ideas to tackle the main problems of regeneration, paving the way to those (descendants) that followed suit. [ABSTRACT FROM AUTHOR]

Graphical abstract Abstract Planarian behavior, physiology, and pattern control offer profound lessons for regenerative medicine, evolutionary biology, morphogenetic engineering, robotics, and unconventional computation. Despite recent advances in the molecular genetics of stem cell differentiation, this model organism's remarkable anatomical homeostasis provokes us with truly fundamental puzzles about the origin of large-scale shape and its relationship to the genome. In this review article, we first highlight several deep mysteries about planarian regeneration in the context of the current paradigm in this field. We then review recent progress in understanding of the physiological control of an endogenous, bioelectric pattern memory that guides regeneration, and how modulating this memory can permanently alter the flatworm's target morphology. Finally, we focus on computational approaches that complement reductive pathway analysis with synthetic, systems-level understanding of morphological decision-making. We analyze existing models of planarian pattern control and highlight recent successes and remaining knowledge gaps in this interdisciplinary frontier field. [ABSTRACT FROM AUTHOR]

Many critical cellular processes, including cell proliferation and tissue repair, exhibit melatonin sensitivity. We examined the effects of exogenous melatonin, photoperiod and temperature on regeneration in Phagocata gracilis, a common North American turbellarian. Planarian regeneration involves the proliferation and migration of pluripotent stem cells capable of replacing missing body components. A characteristic mass of cells (blastema) forms during planarian regeneration, and its formation, growth and differentiation provide reliable endpoints for the study of regenerative processes. We tested the effects of exogenous melatonin at two concentrations and three photoperiods on both head and tail regenerates under seasonal diurnal temperature regimes. High temperatures and melatonin treatment increased mortality in P. gracilis, and surviving melatonin-treated subjects exhibited delayed regeneration. Asymmetrical and deficient regeneration was more common in melatonin-treated, high temperature and dark treated subjects. Additionally, high temperature and melatonin induced a novel locomotor dysfunction. [ABSTRACT FROM AUTHOR]

There is a clear need to establish and validate new methodologies to more quickly and efficiently screen chemicals for potential toxic effects, particularly on development. The emergence of alternative animal systems for rapid toxicology screens presents valuable opportunities to evaluate how systems complement each other. In this article, we compare a chemical library of 87-compounds in 2 such systems, developing zebrafish and freshwater planarians, by screening for developmental neurotoxic effects. We show that the systems' toxicological profiles are complementary to each other, with zebrafish yielding more detailed morphological endpoints and planarians more behavioral endpoints. Overall, zebrafish was more sensitive to this chemical library, yielding 86/87 hits, compared with 50/87 hits in planarians. The difference in sensitivity could not be attributed to molecular weight, log  K ow, or the bioconcentration factor. Of the 87 chemicals, 28 had previously been evaluated in mammalian developmental neuro- (DNT), neuro-, or developmental toxicity studies. Of the 28, 20 were hits in the planarian, and 27 were hits in zebrafish. Eighteen of the 28 had previously been identified as DNT hits in mammals and were highly associated with activity in zebrafish and planarian behavioral assays in this study. Only 1 chemical (of 28) was a false negative in both zebrafish and planarian systems. The differences in endpoint coverage and system sensitivity illustrate the value of a dual systems approach to rapidly query a large chemical-bioactivity space and provide weight-of-evidence for prioritization of chemicals for further testing. [ABSTRACT FROM AUTHOR]

There is an increased recognition in the field of toxicology of the value of medium-to-high-throughput screening methods using in vitro and alternative animal models. We have previously introduced the asexual freshwater planarian Dugesia japonica as a new alternative animal model and proposed that it is particularly well-suited for the study of developmental neurotoxicology. In this article, we discuss how we have expanded and automated our screening methodology to allow for fast screening of multiple behavioral endpoints, developmental toxicity, and mortality. Using an 87-compound library provided by the National Toxicology Program, consisting of known and suspected neurotoxicants, including drugs, flame retardants, industrial chemicals, polycyclic aromatic hydrocarbons (PAHs), pesticides, and presumptive negative controls, we further evaluate the benefits and limitations of the system for medium-throughput screening, focusing on the technical aspects of the system. We show that, in the context of this library, planarians are the most sensitive to pesticides with 16/16 compounds causing toxicity and the least sensitive to PAHs, with only 5/17 causing toxicity. Furthermore, while none of the presumptive negative controls were bioactive in adult planarians, 2/5, acetaminophen and acetylsalicylic acid, were bioactive in regenerating worms. Notably, these compounds were previously reported as developmentally toxic in mammalian studies. Through parallel screening of adults and developing animals, planarians are thus a useful model to detect such developmental-specific effects, which was observed for 13 chemicals in this library. We use the data and experience gained from this screen to propose guidelines for best practices when using planarians for toxicology screens. [ABSTRACT FROM AUTHOR]

Retinoic acid is a known morphogen in regulating animal growth and development. Planaria are a key model system for regeneration and their eyes are a morphological marker of anterior differentiation. We explored the requirement for retinoic acid signaling in the regeneration of body parts in the planaria S. mediterranea using an inhibitor of retinoic acid synthesis, diethylaminobenzaldehyde (DEAB). Whole planaria, soaked in DEAB for three days prior to and five days following amputation, produced trunk and tail fragments with defective anterior regeneration. Following regeneration, up to 80% of posterior fragments developed abnormal eyes. The abnormalities included animals without eyes, with only a single eye, with one enlarged eye, or two eyes of different sizes. Eyes were considered to be functional because animals responded to blue laser light with turning behavior. No abnormalities in eye regeneration were observed in side by side vehicle controls. These results suggest that retinoic acid is necessary for normal eye regeneration following injury and supports a previously undocumented signaling role in planaria eye development. [ABSTRACT FROM AUTHOR]

The study is dedicated to an identification of specific FMRF immunoreactivity in planarian (Girardia tigrina) with special attention to the FMRF-like neuropeptides location near muscle filaments. Using of whole mounts in conjunction with a confocal laser scanning microscopy reveals close spatial relationship of FMRF-immunopositive(-IP) nerves and planarian musculature. Morphological data provide an evidence for FMRF-like peptides participation in the innervations of planarian muscles. The results indicate that the larger part of muscular structures and organs (pharynx, mouth opening, body wall, and tail) of planarian body are deeply supplied by FMRF-IP nerve elements. Such observation of the peptidergic FMRF-IP neurons and its processes in vicinity of the muscle fibers can indicate an important role of the neuropeptides in the regulation of planarian muscle function. [ABSTRACT FROM AUTHOR]

Records of cryptic species have continued to emerge in the scientific literature, often revealed by the use of molecular phylogenetic analyses in an integrative taxonomic approach. This study addresses a group of four striped flatworms from the genus Pasipha Ogren & Kawakatsu, showing a pale median stripe on a dark dorsal surface. Based on morphological and molecular analyses from the cytochrome c oxidase subunit I gene (COI), we establish that we are dealing with sibling species that are closely related to P. brevilineata Leal-Zanchet, Rossi & Alvarenga, 2012, a recently described species with a similar colour pattern. Thus, we describe three of the studied flatworms as new species and propose one new unconfirmed candidate species based on molecular data. In addition, sequence analysis revealed 40 nucleotide autapomorphies supporting the species studied herein. Considering anatomical and histological features, the three new species are differentiated from their congeners mainly by details of the copulatory apparatus, such as the occurrence of an epithelium of pseudostratified appearance lining the female atrium and the shape and position of the proximal portion of the prostatic vesicle. [ABSTRACT FROM AUTHOR]

During animal regeneration, cells must organize into discrete and functional systems. We show that self-organization, along with patterning cues, govern progenitor behavior in planarian regeneration. Surgical paradigms allowed the manipulation of planarian eye regeneration in predictable locations and numbers, generating alternative stable neuroanatomical states for wild-type animals with multiple functional ectopic eyes. We used animals with multiple ectopic eyes and eye transplantation to demonstrate that broad progenitor specification, combined with self-organization, allows anatomy maintenance during regeneration. We propose a model for regenerative progenitors involving (i) migratory targeting cues, (ii) self-organization into existing or regenerating eyes, and (iii) a broad zone, associated with coarse progenitor specification, in which eyes can be targeted by progenitors. These three properties help explain how tissues can be organized during regeneration. [ABSTRACT FROM AUTHOR]

Mechanisms underlying anteroposterior body axis differences during adult tissue maintenance and regeneration are poorly understood. Here, we identify that post-translational modifications through the SUMO (Small Ubiquitin-like Modifier) machinery are evolutionarily conserved in the Lophotrocozoan Schmidtea mediterranea. Disruption of SUMOylation in adult animals by RNA-interference of the only SUMO E2 conjugating enzyme Ubc9 leads to a systemic increase in DNA damage and a remarkable regional defect characterized by increased cell death and loss of the posterior half of the body. We identified that Ubc9 is mainly expressed in planarian stem cells (neoblasts) but it is also transcribed in differentiated cells including neurons. Regeneration in Ubc9(RNAi) animals is impaired and associated with low neoblast proliferation. We present evidence indicating that Ubc9-induced regional cell death is preceded by alterations in transcription and spatial expression of repressors and activators of the Hedgehog signaling pathway. Our results demonstrate that SUMOylation acts as a regional-specific cue to regulate cell fate during tissue renewal and regeneration. [ABSTRACT FROM AUTHOR]

The epidermis is essential for animal survival, providing both a protective barrier and cellular sensor to external environments. The generally conserved embryonic origin of the epidermis, but the broad morphological and functional diversity of this organ across animals is puzzling. We define the transcriptional regulators underlying epidermal lineage differentiation in the planarian Schmidtea mediterranea , an invertebrate organism that, unlike fruitflies and nematodes, continuously replaces its epidermal cells. We find that Smed-p53 , Sox and Pax transcription factors are essential regulators of epidermal homeostasis, and act cooperatively to regulate genes associated with early epidermal precursor cell differentiation, including a tandemly arrayed novel gene family ( prog ) of secreted proteins. Additionally, we report on the discovery of distinct and previously undescribed secreted organelles whose production is dependent on the transcriptional activity of soxP-3 , and which we term Hyman vesicles. [ABSTRACT FROM AUTHOR]

Many pigment cells acquire unique structural properties and gene expression profiles during animal development. The underlying differentiation pathways have been well characterized in cells formed during embryogenesis, such as the neural crest-derived melanocyte. However, much less is known about the developmental origins of pigment cells produced in adult organisms during tissue homeostasis and repair. Here we report a lineage analysis of ommochrome- and porphyrin-producing cells in the brown, freshwater planarian Schmidtea mediterranea. Using an RNAsequencing approach, we identified two classes of markers expressed in sequential fashion when new pigment cells are generated during regeneration or in response to pigment cell ablation. We also report roles for FOXF-1 and ETS-1 transcription factors, as well as for an FGFR-like molecule, in the specification and maintenance of this cell type. Together, our results provide insights into mechanisms of adult pigment cell development in the strikingly colorful Platyhelminthes phylum. [ABSTRACT FROM AUTHOR]

Methylisothiazolinone (MIT) is a common biocide used in cosmetic and industrial settings. Studies have demonstrated that MIT is a human sensitizer, to the extent that in 2013 MIT was named allergen of the year. Recently, we showed that MIT exposure in Xenopus laevis (the African clawed frog) inhibits wound healing and tail regeneration. However, it is unknown whether MIT affects these processes in other animals. Here, we investigated the effects of MIT exposure in planaria—non-parasitic freshwater flatworms able to regenerate all tissues after injury. Using a common research strain of Dugesia japonica , we determined that intact planarians exposed to 15 μM MIT displayed both neuromuscular and epithelial-integrity defects. Furthermore, regenerating (head and tail) fragments exposed to 15 μM MIT failed to close wounds or had significantly delayed wound healing. Planarian wounds normally close within 1 h after injury. However, most MIT-exposed animals retained open wounds at 24 h and subsequently died, and those few animals that were able to undergo delayed wound healing without dying exhibited abnormal regeneration. For instance, head regeneration was severely delayed or inhibited, with anterior structures such as eyes failing to form in newly produced tissues. These data suggest that MIT directly affects both wound healing and regeneration in planarians. Next, we investigated the ability of thiol-containing antioxidants to rescue planarian wound closure during MIT exposure. The data reveal both n -acetyl cysteine and glutathione were each able to fully rescue MIT inhibition of wound healing. Lastly, we established MIT toxicity levels by determining the LC 50 of 5 different planarian species: D. japonica, Schmidtea mediterranea , Girardia tigrina, Girardia dorotocephala, and Phagocata gracilis . Our LC 50 data revealed that concentrations as low as 39 μM (4.5 ppm) are lethal to planarians, with concentrations of just 5 μM inhibiting wound healing, and suggest that phylogeny is predictive of species toxicity levels. Together these results indicate MIT may have broad wound healing effects on aquatic species in general and are not limited to X. laevis alone. Future studies should investigate the impact of MIT on wound healing in other organisms, including non-aquatic organisms and mammals. [ABSTRACT FROM AUTHOR]

Planarians, which are non-parasitic flatworms, are highly resistant to bacterial infections. To better understand the mechanisms underlying this resistance, we investigated the role of the circadian machinery in the anti-bacterial response of the freshwater planarian Schmidtea mediterranea. We identified Smed-Tim from S. mediterranea as a homolog of the mammalian clock gene Tim. We showed via RNA interference that Smed-Tim is required for the antimicrobial activities of Schmidtea mediterranea against Staphylococcus aureus infection during the light/dark cycle. Indeed, S. aureus infection leads to the expression of Smed-Tim, which in turn promotes Smed-Traf6 and Smed-morn2, but not Smed-p38 MAPK expression, 2 master regulators of planarian anti-microbial responses. [ABSTRACT FROM AUTHOR]

Regeneration of complex anatomical structures is an emergent phenomenon arising from complex interplay between various underlying cellular components and processes. It is still largely unclear how coordination among cells leads to accurate regeneration of the organism, preserving the location, shape, and composition of the parts with respect to the whole. Here, we examine at the global interaction of cells in a computational model of planarian regeneration. A key feature of our model is the integration of multiple organizational levels of an organism – from cells, to network, to global shape. The computational model is able to replicate most of the experimental observations thereby facilitating study of the putative mechanisms. We observe that a hierarchical interplay between local and long range signaling acts as a positional map that guides the cellular fate at any position. Furthermore, we have quantified the quality of regeneration using a metric that provides a sense of how well the regenerated organism resembles its original shape. Planaria are complex bilaterian animals with a remarkable capability. They regenerate after almost any sort of surgical damage, re-growing precisely the needed structures at the correct locations and stopping growth and remodeling after their target morphology is complete. Understanding the dynamic regulation of biological patterning is key to transformative advances in regenerative medicine and synthetic bioengineering. Here, we propose a connectionist model of pattern memory during planarian regeneration. [ABSTRACT FROM AUTHOR]

• Bioelectrical patterns based on multicellular mean-field potential are morphologically relevant. • We study theoretically the transplantation of fragments from different planarian species. • The model provides biological insights on single-cell states coupled by the intercellular connectivity. • The simulations predict that small but timely bioelectrical actions may have effects on the head shape. Head-tail planaria morphologies are influenced by the electric potential differences across the animal's primary axis, as evidenced e.g. by voltage-sensitive dyes and functional experiments that create permanent lines of 2-headed but genetically wild-type animals. However, bioelectrical and biochemical models that make predictions on what would happen in the case of spatial chimeras made by tissue transplantation from different planaria (different species and head shapes) are lacking. Here, we use a bioelectrical model to qualitatively describe the effects of tissue transplantation on the shape of the regenerated head. To this end, we assume that the cells may have distinct sets of ion channels and ascribe the system outcome to the axial distributions of average cell potentials over morphologically relevant regions. Our rationale is that the distributions of signaling ions and molecules are spatially coupled with multicellular electric potentials. Thus, long-time downstream transcriptional events should be triggered by short-time bioelectrical processes. We show that relatively small differences between the ion channel characteristics of the cells could eventually give noticeable changes in the electric potential profiles and the expected morphological deviations, which suggests that small but timely bioelectrical actions may have significant morphological effects. Our approach is based on the observed relationships between bioelectrical regionalization and biochemical gradients in body-plan studies. Such models are relevant to regenerative, developmental, and cancer biology in which cells with distinct properties and morphogenetic target states confront each other in the same tissue. [ABSTRACT FROM AUTHOR]