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Keller, L.

Publications and source records attributed to Keller, L..

At least 19 recordsLinked to original sources

Using a wearable EEG device to examine age trends in sleep macro- and micro-architecture across adolescence

Adolescence is a period of distinct maturational changes in sleep physiology. Age-related trends in sleep physiology have been captured using laboratory-based polysomnography, a method limited by logistical burden and high cost. We tested the ability of the accessible Dreem3 sleep EEG headband to replicate established age effects in sleep physiology from late childhood through early adulthood. Typically developing youth (N=100, 9-26 years) completed 3-4 consecutive nights of at-home sleep recording. We estimated age-related trends across eight macro-architecture and 15 micro-architecture variables with known age effects, and conducted exploratory analyses of 24 additional variables. Dreem replicated established age trends, including increases in non-rapid eye movement (NREM) stage 2, and decreases in N3, time in bed, NREM delta and theta power with increasing age. Exploratory analysis revealed age effects in twelve variables, including decreases in spindle activity with increasing age. Sleep EEG wearables offer an accessible way to characterize sleep physiology development.

neuroscience↗

Social and Spatial Affinity Drive Wound Care by Ants

In group-living animals, open wounds pose a risk not only for the injured individual but also for the entire group, especially when the risk of infection transmission is high. To mitigate this danger, different species have developed diverse strategies - with ants representing one of the few taxa beyond humans that engage in social wound care. Yet, who provides this care and what determines its allocation remains unclear. To answer these questions, we combined controlled injury experiments with automated behavioral tracking across six colonies of the ant Camponotus fellah. Our results reveal that wound care is primarily provided by a non-specialized subset of individuals whose behavioral traits align with a transitional phase between the nurse and forager roles. Among these transitioning ants, care provision was best predicted by their prior spatial and social proximity to the injured individuals in the hours before the injury, indicating that pre-existing social affinity influences caregiving behavior. These findings offer the first empirical evidence of how caregivers are integrated into the colonys social structure and highlight the role of social interactions in shaping the organization of wound care in ants.

animal behavior and cognition↗

A blastocyst-derived in vitro model of the human chorion

The placenta supports the foetus by mediating nutrient and gas exchange, hormone production, and immune protection. Yet, human placental biology remains poorly understood due to limited in vitro models. The foetal part of the placenta, the chorion, consists of a trophoblast-derived layer and a mesenchymal zone containing extraembryonic mesoderm cells. Here, we show that human blastocysts can give rise to both trophoblast organoids and extraembryonic mesoderm cells under the same culture conditions. Trophoblast organoids originate from both outer trophoblast cells and inner cells of the blastocyst, while extraembryonic mesoderm cells derive exclusively from inner cells. These organoids recapitulate the cellular composition, morphology, and function of the in vivo trophoblast. Moreover, given the high rate of aneuploidy at the blastocyst stage, aneuploid trophoblast organoids can be readily generated. These models reflect the trophoblasts unique ability to tolerate aneuploidy and offer a valuable platform to study human placental development and pathophysiology.

developmental biology↗

An abundance of aliC and aliD genes were identified in saliva using a novel multiplex qPCR to characterize group II non-encapsulated pneumococci with improved specificity

BackgroundSurveillance of pneumococcus is reporting increasing prevalence of non-encapsulated pneumococci (NESp). NESp are an important reservoir for genetic exchange among streptococci, including for antimicrobial resistance (AMR), and are increasingly implicated in disease. Disease-associated NESp commonly carry the virulence genes pspK, or aliC and aliD in their cps locus instead of capsule genes. While molecular methods targeting the cps region are widely used for serotyping encapsulated strains, there are few assays available for classification of NESp, meaning it is not widely undertaken. Therefore, we exploited these genes as targets for a novel qPCR assay for detecting and classifying NESp strains with improved efficiency and specificity. MethodsWe conducted bioinformatic analysis on sequences from 30 NESp and 23 other mitis-group streptococcal sequences and developed a multiplex-qPCR, targeting pspK, aliD and two regions of aliC. The assay was validated using 11 previously characterised, and 5 uncharacterised NESp isolates. We then applied the assay to DNA extracted from culture-enriched saliva, and isolated and characterised suspected NESp colonies, with confirmation by whole genome sequencing. ResultsBioinformatic analyses demonstrated that previously published primers for aliC and aliD had low pneumococcal-specificity but indicated that targeting two regions of aliC would improve species-specificity, without compromising sensitivity. Our novel multiplex assay accurately typed all isolates. When screening saliva, we found a high prevalence of aliC and aliD, even in samples negative for pneumococcal genes lytA and piaB. Isolated colonies which were aliC and aliD positive could be differentiated as non-pneumococcal streptococci using our assay. ConclusionOur multiplex-qPCR assay can be used to efficiently screen even highly polymicrobial samples, such as saliva, for NESp genes, to detect and differentiate potentially pathogenic NESp clades from closely related mitis-group streptococci. This will allow for a better understanding of the true prevalence of NESp, and their impact upon pneumococcal carriage, disease, and AMR.

microbiology↗

Ultrastructural, metabolic and genetic determinants of the acquisition of macrolide resistance by Streptococcus pneumoniae

AimStreptococcus pneumoniae (Spn) acquires genes for macrolide resistance, MEGA or ermB, in the human host. These genes are carried either in the chromosome, or on integrative conjugative elements (ICEs). Here, we investigated molecular determinants of the acquisition of macrolide resistance. Methods and ResultsWhole genome analysis was conducted for 128 macrolide-resistant pneumococcal isolates to identify the presence of MEGA (44.5%, 57/128) or ermB (100%), and recombination events in Tn916-related elements or in the locus comCDE encoding competence genes. Confocal and electron microscopy studies demonstrated that, during the acquisition of macrolide resistance, pneumococcal strains formed clusters of varying size, with the largest aggregates having a median size of [~]1600 {micro}m2. Remarkably, these pneumococcal aggregates comprise both encapsulated and nonencapsulated pneumococci, exhibited physical interaction, and spanned extracellular and intracellular compartments. We assessed the recombination frequency (rF) for the acquisition of macrolide resistance by a recipient D39 strain, from pneumococcal strains carrying MEGA ([~]5.4 kb) in the chromone, or in large ICEs (>23 kb). Notably, the rF for the acquisition of MEGA, whether in the chromosome or carried on an ICE was similar. However, the rF adjusted to the acquisition of the full-length ICE ([~]52 kb), compared to that of the capsule locus ([~]23 kb) that is acquired by transformation, was three orders of magnitude higher. Finally, metabolomics studies revealed a link between the acquisition of ICE and the metabolic pathways involving nicotinic acid and sucrose. ConclusionsExtracellular and intracellular pneumococcal clusters facilitate the acquisition of full-length ICE at a rF higher than that of typical transformation events, involving distinct metabolic changes that present potential targets for interventions.

microbiology↗

Gut microbiota influences foraging onset without affecting division of labor and associated physiological hallmarks in honeybees

Gut microbes can impact cognition and behavior, but whether they regulate division of labor in animal societies is unknown. We addressed this question using honeybees since they exhibit division of labor between nurses and foragers and because their gut microbiota can be manipulated. Using automated behavioral tracking and controlling for co-housing effects, we show that gut microbes influence the age at which bees start foraging but have no effects on the time spent in a foraging area and number of foraging trips. Moreover, the gut microbiota did not influence hallmarks of behavioral maturation such as body weight, cuticular hydrocarbon (CHC) profile, hypopharyngeal gland size, and the proportion of bees maturing into foragers. Overall, this study shows that the honeybee gut microbiota does not affect division of labor but rather plays an important function in controlling the onset of bee foraging.

evolutionary biology↗

Opportunity nest egg: insights on the nutritional ecology, life history and captive management of three species of kiwi (Apteryx spp.) chick from Operation Nest Egg zoo hand-rearing records.

Zoo data collected by keepers while looking after endangered species are increasingly recognised as important scientific resources. As chicks, New Zealands threatened kiwi (Apteryx spp.) are subject to the protective conservation programme Operation Nest Egg (ONE), during which growth, developmental and life history data are recorded. We have conducted comparative analyses on hand rearing records from 306 chicks, from Coromandel, Eastern and Western populations of Brown kiwi (A. mantelli), and rowi (A. rowi) and Haast tokoeka (A. australis "Haast"). We analysed chick responses to an old diet vs. a new kiwi diet, introduced in 2017. Both diets are fixed nutrient ratio mixtures. The old diet was high-protein, low-energy, while the new diet is high-fat, high-energy, with better micronutrient levels. We found Coromandel chicks, who originate from the environmentally variable K[u]aotunu Peninsula, grow the most efficiently overall on either diet, indicating they may be nutritional generalists. Western and Eastern chicks growth efficiency was intermediate, while the South Island species grew the least efficiently on either diet. Rowi chicks developed the fastest overall on either diet, while Haast tokoeka chicks developed the most slowly, especially on the new diet. Rowi chicks therefore had to eat large volumes of either diet over a short time to maintain their rapid development, while Haast chicks were required to eat large volumes, especially of the new diet, over a protracted developmental period. This situation may have led South Island chicks to over-consume one or another diet component, with likely health consequences. Neither diet was obviously superior for chick wellbeing overall, though the new diet better supported chicks that needed hand feeding. This work demonstrates different genetic populations of kiwi differ in their physiological responses to nutrition. As ONE is ongoing, tailored diets for chicks from each genetic group should be developed, and we present methods to achieve this. In our life history trait analyses, we found chick starting size (hatch mass) did not significantly influence growth efficiency across kiwi genetic group, nor did chick sex. We identified that chicks malpositioned as embryos were more likely to require extended periods of hand feeding, and that Eastern males produce more malpositioned embryos than other populations. Our study shows that effective zoo records can be used to improve captive care, to stimulate future research to refine species management practices, and to explore fundamental questions of life history evolution in wild and captive populations.

zoology↗

Trophic eggs affect caste determination in the ant Pogonomyrmex rugosus

Understanding how a single genome creates distinct phenotypes remains a fundamental challenge for biologists. Social insects provide a striking example of polyphenism, with queen and worker castes exhibiting morphological, behavioural, and reproductive differences. Here we show that trophic eggs, which do not contain an embryo and are primarily regarded as a source of food, play a role in the process of caste determination in the harvester ant Pogonomyrmex rugosus. When first instar larvae were given access to trophic eggs, they mostly developed into workers. By contrast, larvae without access to trophic eggs developed into queens. We found that trophic eggs differ in many ways from viable eggs, including texture, morphology and their contents of protein, triglycerides, glycogen, sugar and small RNAs. Moreover, comparison of miRNA fragment size distributions suggests differences in the composition of miRNAs between the two egg types. This is the first demonstration of trophic eggs playing a role in caste determination in social insects.

evolutionary biology↗

Natural alleles at the Doa locus underpin evolutionary changes in Drosophila lifespan and fecundity

Evolve and resequence (E&R) studies in Drosophila melanogaster have identified many candidate loci underlying the evolution of ageing and life history, but experiments that validate the effects of such candidates remain rare. In a recent E&R study we have identified several alleles of the LAMMER kinase Darkener of apricot (Doa) as candidates for evolutionary changes in lifespan and fecundity. Here, we use two complementary approaches to confirm a functional role of Doa in life-history evolution. First, we used transgenic RNAi to study the effects of Doa at the whole-gene level. Ubiquitous silencing of expression in adult flies reduced both lifespan and fecundity, indicating pleiotropic effects. Second, to characterize segregating variation at Doa, we examined four candidate single nucleotide polymorphisms (SNPs; Doa-1, -2, -3, -4) using a genetic association approach. Three candidate SNPs had effects that were qualitatively consistent with expectations based on our E&R study: Doa-2 pleiotropically affected both lifespan and late-life fecundity; Doa-1 affected lifespan (but not fecundity); and Doa-4 affected late-life fecundity (but not lifespan). Finally, the last candidate allele (Doa-3) also affected lifespan, but in the opposite direction than predicted.

evolutionary biology↗

Spatial expression of an mRNA encoding Tie2-agonist in the capillary endothelium of the lung prevents pulmonary vascular leakage

Angiopoietin ligands Ang1 and Ang2 and the Tie2 receptor tyrosine kinases form an endothelial signaling pathway regulating vascular homeostasis and controlling vessel permeability, inflammation and angiogenic responses. Whereas Ang1-mediated Tie2 activation reduces inflammation and endothelial permeability, its antagonist, Ang2 increases it. Increased plasma Ang2 levels are associated with poor outcomes in patients with acute lung injury (ALI), as well as in acute respiratory distress syndrome (ARDS). In the study presented here we tested the effect of a novel synthetic, nucleoside-modified mRNA-76 encoding for a hyperactive Ang1 derived fusion protein (COMP-Ang1) on attenuating post-inflammation vascular leakage. COMP-Ang1 mRNA was formulated into a cationic lipid nanoparticle (cLNP) using an optimized mixture of three different lipids and a microfluidic mixing technology. After intravenous injection, the respective mRNA-loaded LNPs were found to be delivered predominantly to the endothelial cells of the lung, while sparing other vascular beds. Also, the specific multimeric folding of the COMP-Ang1 protein complex appeared to be pivotal for its activity in preventing vascular leakage and in restoring the alveolar-endothelial barrier function in the inflamed and injured pulmonary vasculature. The mode of action of mRNA-76, such as its activation of the Tie2 signal transduction pathway, was tested by pharmacological studies in vitro and in vivo by systemic administration in respective mouse models. mRNA-76 was found to prevent lung vascular leakage/lung edema as well as neutrophil infiltration in an LPS-challenging model.

molecular biology↗

The evolution of behavioral cues and signaling in displaced communication

Displaced communication, whereby individuals communicate regarding a subject that is not immediately present (spatially or temporally), is one of the key innovations of human language. It also occurs in a few animal species, most notably the honeybee, where the waggle dance is used to communicate the location and quality of a patch of flowers. However, it is difficult to study how it emerged given the paucity of species displaying this capacity and the fact that it often occurs via complex multimodal signals. To address this issue, we developed a novel paradigm in which we conducted experimental evolution with foraging agents endowed with neural networks that regulate their movement and the production of signals. Displaced communication readily evolved but, surprisingly, agents did not use signal amplitude to convey information on food location. Instead, they used signal onset-delay and duration-based mode of communication, which depends on the motion of the agent within a communication area. When agents were experimentally prevented from using these modes of communication, they evolved to use signal amplitude instead. Interestingly, this mode of communication was more efficient and led to higher performance. Subsequent controlled experiments suggested that this more efficient mode of communication failed to evolve because it took more generations to emerge than communication grounded on the onset-delay and length of signaling. These results reveal that displaced communication is likely to initially evolve from non-communicative behavioral cues providing incidental information with evolution later leading to more efficient communication systems through a ritualization process. Author SummaryThe evolution of displaced communication, the process through which individuals share information about a remote object (in space or time), is a key innovation in language. By conducting experimental evolution we found that displaced communication is more likely to leverage and evolve from behavioural cues, such as the agents movement, rather than from dedicated communication modes, such as the amplitude of emitted signals. This phenomenon is shown to happen because communication via signal amplitude -although more efficient-is slower to evolve. The simple behaviors and neural networks of the agents studied here, also suggest that communication may evolve more frequently than expected via ritualization, a process whereby an action or behavior pattern in an animal loses its original function but is retained for its role in display or other social interactions.

evolutionary biology↗

Infection signaling and antimicrobial wound care in an ant society

Infected wounds pose a major mortality risk in animals1,2. Injuries are common in the ant Megaponera analis, which raids pugnacious prey3,4. Here we show that M. analis can determine when wounds are infected and treat them accordingly. By applying a variety of antimicrobial compounds and proteins secreted from the metapleural gland to infected wounds, workers reduce the mortality of infected individuals by 90%. Chemical analyses showed that wound infection is associated with specific changes in the cuticular hydrocarbon profile, thereby likely allowing nestmates to diagnose the infection state of injured individuals and apply the appropriate antimicrobial treatment. This study demonstrates that the targeted use of antimicrobials to treat infected wounds, previously thought to be a uniquely human behavior, has evolved in insect societies as well.

animal behavior and cognition↗

Invasion genetics of the longhorn crazy ant: the global expansion of a double-clonal reproduction system

Reproduction mode represents a key determinant for success of biological invasion as it influences the genetic variation and evolutionary potential of introduced populations. The worlds most widespread invasive ant, Paratrechina longicornis, was found to display an unusual double-clonal reproduction system, whereby both males and queens were produced clonally, while workers are produced sexually. Despite its worldwide distribution, the origin of this ant species and the prevalence of the double-clonal reproductive system across the ants geographic range remain unknown. To retrace the evolutionary history of this global invasive species and its reproductive system, we examined genetic variation and characterized the mode of reproduction of P. longicornis sampled worldwide using both microsatellite genotyping and mitochondrial DNA sequencing approaches. Analyses of global genetic variations indicate that the Indian subcontinent is a genetic diversity hotspot of this species, suggesting that this geographic area is at least part of its native range. Our analyses revealed that inferred native and introduced populations both exhibit double-clonal reproduction. Remarkably, queens and males worldwide belong to two separate, non-recombining clonal lineages. Workers are highly heterozygous and first-generation inter- lineage hybrids, a pattern strongly supportive of a strict worldwide prevalence of double clonality. By maintaining heterozygosity in the worker force, this unusual genetic system allows P. longicornis to avoid inbreeding during colonization bottlenecks and may have acted as an adaptive trait linked to the species invasion success.

evolutionary biology↗

The gut microbiota affects the social network of honeybees

The gut microbiota influences animal neurophysiology and behavior but has not previously been documented to affect emergent group-level behaviors. Here we combine gut microbiota manipulation with automated behavioral tracking of honeybee sub-colonies to show that the microbiota increases the rate and specialization of social interactions. Microbiota colonization was associated with higher abundances of one third of metabolites detected in the brain, including several amino acids, and a subset of these metabolites were significant predictors of social interactions. Colonization also affected brain transcriptional processes related to amino acid metabolism and epigenetic modification in a brain region involved in sensory perception. These results demonstrate that the gut microbiota modulates the emergent colony social network of honeybees, likely via changes in chromatin accessibility and amino acid biosynthesis.

animal behavior and cognition↗

A single nucleotide variant in the PPARgamma-homolog Eip75B affects fecundity in Drosophila

Single nucleotide polymorphisms are the most common type of genetic variation, but how these variants contribute to the evolutionary adaptation of complex phenotypes is largely unknown. Experimental evolution and genome-wide association studies have demonstrated that variation in the PPARg-homolog Eip75B is associated with longevity and life-history differences in the fruit fly Drosophila melanogaster. Using RNAi knockdown, we first demonstrate that reduced expression of Eip75B in adults affects lifespan, egg-laying rate and egg volume. We then tested the effect of a naturally occurring SNP variant within a cis-regulatory domain of Eip75B by applying two complementary approaches: a Mendelian randomization approach using lines of the Drosophila Genetic Reference Panel, and allelic replacement using precise CRISPR/Cas9-induced genome editing. Our experiments reveal that this natural polymorphism has a significant pleiotropic effect on fecundity and egg-to-adult viability, but not on longevity or other life-history traits. These results provide a rare functional validation at the nucleotide level and identify a natural allelic variant affecting fitness and life-history adaptation.

evolutionary biology↗

Pdap1 protects mature B lymphocytes from stress-induced cell death and promotes antibody gene diversification

The establishment of protective humoral immunity is dependent on the ability of mature B cells to undergo antibody gene diversification while adjusting to the physiological stressors induced by activation with the antigen. Mature B cells diversify their antibody genes by class switch recombination (CSR) and somatic hypermutation (SHM), which are both dependent on efficient induction of activation-induced cytidine deaminase (AID). Here, we identified PDGFA-associated protein 1 (Pdap1) as an essential regulator of cellular homeostasis in mature B cells. Pdap1 deficiency leads to sustained expression of the integrated stress response (ISR) effector activating transcription factor 4 (Atf4) and induction of the ISR transcriptional program, increased cell death, and defective AID expression. As a consequence, loss of Pdap1 reduces germinal center B cell formation and impairs CSR and SHM. Thus, Pdap1 protects mature B cells against chronic ISR activation and ensures efficient antibody diversification by promoting their survival and optimal function.

immunology↗

Antisense oligonucleotide therapy rescues aggresome formation in a novel Spinocerebellar Ataxia type 3 human embryonic stem cell line.

Spinocerebellar ataxia type 3 (SCA3) is a fatal, late-onset neurodegenerative disorder characterized by selective neuropathology in the brainstem, cerebellum, spinal cord, and substantia nigra. Here, we characterize the first NIH-approved human embryonic stem cell (hESC) line derived from an embryo harboring the SCA3 mutation. Referred here as SCA3-hESC, this line is heterozygous for the mutant polyglutamine-encoding CAG repeat expansion in the ATXN3 gene within the pathogenic repeat range for SCA3. We observed relevant molecular hallmarks of the human disease at all differentiation stages from stem cells to cortical neurons, including robust ATXN3 aggregation and altered expression of key components of the protein quality control machinery. Finally, antisense oligonucleotide-mediated reduction of ATXN3 prevented the formation of p62-positive aggresomes in SCA3-hESCs. The SCA3-hESC line offers a unique and highly relevant human disease model that holds strong potential to advance understanding of SCA3 disease mechanisms and facilitate the evaluation of possible SCA3 therapies.\n\nHighlightsO_LIGenerated first NIH-approved SCA3 human embryonic stem cell line (SCA3-hESC)\nC_LIO_LISCA3-hESC exhibit robust ATXN3 aggregation pathology and form p62+ aggresomes\nC_LIO_LIAnti-ATXN3 antisense oligonucleotides rescue aggresome formation in SCA3-hESC\nC_LIO_LIDerived SCA3 neurons form aggregates and exhibit impaired protein quality control\nC_LI

neuroscience↗

Individuals' expected genetic contributions to future generations, reproductive value, and short-term metrics of fitness in free-living song sparrows (Melospiza melodia)

Appropriately defining and enumerating fitness is fundamental to explaining and predicting evolutionary dynamics. Yet theoretical concepts of fitness are often hard to translate into quantities that can be quantified in wild populations experiencing complex environmental, demographic, genetic and selective variation. While the fittest entities might be widely understood to be those that ultimately leave most descendants at some future time, such long-term legacies are hard to measure, impeding evaluation of how well more tractable short-term metrics of individual fitness directly predict longer-term outcomes. One opportunity for conceptual and empirical convergence stems from the principle of individual reproductive value (Vi), defined as the number of copies of each of an individuals alleles that is expected to be present in future generations given the individuals realised pedigree of descendants. Since Vi tightly predicts an individuals longer-term genetic contribution, quantifying Vi provides a tractable route to quantifying what, to date, has been an abstract fitness concept. We used complete pedigree data from free-living song sparrows (Melospiza melodia) to demonstrate that individuals expected genetic contributions stabilise within an observed 20-year time period, allowing individual Vi to be evaluated. Considerable among-individual variation in Vi was evident in both sexes. However, standard short-term metrics of individual fitness, comprising lifespan, lifetime reproductive success and projected growth rate, typically explained less than half the variation. Given these results, we discuss what evolutionary inferences can and cannot be directly drawn from short-term versus longer-term fitness metrics observed on individuals, and highlight that analyses of pedigree structure may provide useful complementary insights into evolutionary processes and outcomes.

evolutionary biology↗