Biology subjects

Smith, D. R.

Publications and source records attributed to Smith, D. R..

26 records · Page 2Linked to original sources

bioRxiv2022

Plastomes of Hyalomonas oviformis and Hyalogonium fusiforme evolved dissimilar architecture after loss of photosynthesis

The loss of photosynthesis in land plants and algae is typically associated with parasitism but can also occur in free-living species, including chlamydomonadalean green algae. The plastid genomes (ptDNAs) of colorless chlamydomonadalean species are surprisingly diverse in architecture, including highly expanded forms (Polytoma uvella, Leontynka pallida) as well as outright genome loss (Polytomella species). Here, we explore the ptDNAs of Hyalomonas (Hm.) oviformis (SAG 62-27; formerly known as Polytoma oviforme) and Hyalogonium (Hg.) fusiforme (SAG 62-1c), each representing independent losses of photosynthesis within the Chlamydomonadales. The Hm. oviformis ptDNA is moderately sized (132 kb), smaller than that of its photosynthetic relative Hyalomonas chlamydogama SAG 11-48b (198.3 kb), with a reduced gene complement but still encoding the ATPase subunits. The Hg. fusiforme plastome, however, is the largest yet observed in colorless plants or algae (~463 kb) and has a coding repertoire that is almost identical to that of its photosynthetic relatives in the genus Chlorogonium. Furthermore, the ptDNA of Hg. fusiforme shows no clear evidence of pseudogenization, which is consistent with our analyses showing that Hg. fusiforme is the non-photosynthetic lineage of most recent origin among the known colorless Chlamydomonadales. Together, these new ptDNAs clearly show that, in contrast to parasitic algae, plastid genome compaction is not an obligatory route following the loss of photosynthesis in free-living algae, and that certain chlamydomonadalean algae have a remarkable propensity for genomic expansion, which can persist regardless of the trophic strategy. One sentence summaryThe plastid genomes of two free-living chlamydomonadalean algae, Hyalomonas oviformis and Hyalogonium fusiforme, reveal different evolutionary stages following the loss of photosynthesis.

genomics↗

bioRxiv2022

Activity of the manganese efflux transporter SLC30A10 in dopaminergic but not GABAergic neurons protects against neurotoxicity

Metals such as copper, iron, and manganese (Mn) are essential for life, but induce neurotoxicity at elevated levels. Yet, the neuronal mechanisms of metal-induced neurological disease are largely unclear. A primary limitation has been an inability to selectively alter metal levels in specific neurons, so that the role of the targeted neurons in disease biology can be isolated. Here, we show that neuron-specific depletion of metal efflux transporters provides the feasibility to overcome this limitation by focusing on Mn, which accumulates in the basal ganglia and induces motor disease, and the Mn-specific efflux transporter SLC30A10. Pan-neuronal/glial Slc30a10 knockout mice exhibited increased basal ganglia Mn levels and hypolocomotor deficits in early-life (pre-adulthood), which were exacerbated by Mn exposure. The locomotor deficits of the pan-neuronal/glial strain was associated with a reduction in evoked striatal dopamine release without dopaminergic (DAergic) neurodegeneration or changes in striatal tissue dopamine levels. Furthermore, DAergic-specific, but not GABAergic-specific, Slc30a10 knockout mice recapitulated the hypolocomotor phenotype of the pan-neuronal/glial knockouts in early-life although Mn levels were elevated in the targeted basal ganglia regions of both the neuron-specific strains. Put together, our results imply that (1) activity of SLC30A10 in DAergic neurons is necessary to protect against early-life Mn neurotoxicity; (2) increasing Mn levels in DAergic neurons is sufficient to induce early-life motor disease, suggesting that Mn targets DAergic neurons in the early-life period to induce motor deficits; and (3) neuron-specific knockout of metal efflux transporters may be a widely applicable strategy to elucidate mechanisms of metal-induced neurotoxicity.

neuroscience↗

bioRxiv2022

Contribution of spontaneous mutations to quantitative and molecular variation at the highly repetitive rDNA locus in yeast

The ribosomal DNA array in Saccharomyces cerevisiae consists of many tandem repeats whose copy number is believed to be functionally important but highly labile. Regulatory mechanisms have evolved to maintain copy number by directed mutation, but how spontaneous variation at this locus is generated and selected has not been well characterized. We applied a mutation accumulation approach to quantify the impacts of mutation and selection on this unique genomic feature across hundreds of mutant strains. We find that mutational variance for this trait is relatively high, and that un-selected mutations throughout the genome can disrupt copy number maintenance. In consequence, copy number generally declines, consistent with a model of regulation where copy number is normally increased when low. This pattern holds across ploidy levels and strains in the standard lab environment, but differs under some stressful conditions. We identify several alleles, gene categories and genomic features that likely affect copy number, including aneuploidy for chromosome XII. Copy number change is associated with reduced growth in diploids, consistent with stabilizing selection. Levels of standing variation in copy number are well predicted by a balance between mutation and stabilizing selection, suggesting this trait is not subject to strong diversifying selection in the wild. The rate and spectrum of point mutations within the rDNA locus itself is distinct from the rest of the genome, and predictive of polymorphism locations. Our findings help differentiate the roles of mutation and selection and indicate that spontaneous mutation patterns shape several aspects of ribosomal DNA evolution.

genetics↗

bioRxiv2020

The Utilization of Advance Telemetry to Investigate Important Physiological Parameters Including Electroencephalography in Cynomolgus Macaques Following Aerosol Challenge with Eastern Equine Encephalitis Virus

Most alphaviruses are mosquito-borne and can cause severe disease in humans and domesticated animals. In North America, eastern equine encephalitis virus (EEEV) is an important human pathogen with case fatality rates of 30-90%. Currently, there are no therapeutics or vaccines to treat and/or prevent human infection. One critical impediment in countermeasure development is the lack of insight into clinically relevant parameters in a susceptible animal model. This study examined the disease course of EEEV in a cynomolgus macaque model utilizing advanced telemetry technology to continuously and simultaneously measure temperature, respiration, activity, heart rate, blood pressure, electrocardiogram (ECG), and electroencephalography (EEG) following an aerosol challenge at 7.0 log10 PFU. Following challenge, all parameters were rapidly and substantially altered with peak alterations from baseline ranged as follows: temperature (+3.0-4.2 {degrees}C), respiration rate (+56-128%), activity (+15-76% daytime and +5-22% nighttime), heart rate (+67-190%), systolic (+44-67%) and diastolic blood pressure (+45-80%). Cardiac abnormalities comprised of alterations in QRS and PR duration, QTc Bazett, T wave morphology, amplitude of the QRS complex, and sinoatrial arrest. An unexpected finding of the study was the first documented evidence of a critical cardiac event as an immediate cause of euthanasia in one NHP. All brain waves were rapidly (12-24 hpi) and profoundly altered with increases of up to 6,800% and severe diffuse slowing of all waves with decreases of [~]99%. Lastly, all NHPs exhibited disruption of the circadian rhythm, sleep, and food/fluid intake. Accordingly, all NHPs met the euthanasia criteria by [~]106-140 hpi. This is the first of its kind study utilizing state of the art telemetry to investigate multiple clinical parameters relevant to human EEEV infection in a susceptible cynomolgus macaque model. The study provides critical insights into EEEV pathogenesis and the parameters identified will improve animal model development to facilitate rapid evaluation of vaccines and therapeutics. Author SummaryIn North America, EEEV causes the most severe mosquito-borne disease in humans highlighted by fatal encephalitis and permeant debilitating neurological sequelae in survivors. The first confirmed human cases were reported more than 80 years ago and since then multiple sporadic outbreaks have occurred including one of the largest in 2019. Unfortunately, most human infections are diagnosed at the on-set of severe neurological symptoms and consequently a detailed disease course in humans is lacking. This gap in knowledge is a significant obstacle in the development of appropriate animal models to evaluate countermeasures. Here, we performed a cutting-edge study by utilizing a new telemetry technology to understand the course of EEEV infection in a susceptible macaque model by measuring multiple physiological parameters relevant to human disease. Our study demonstrates that the infection rapidly produces considerable alterations in many critical parameters including the electrical activity of the heart and the brain leading to severe disease. The study also highlights the extraordinary potential of new telemetry technology to develop the next generation of animal models in order to comprehensively investigate pathogenesis as well as evaluate countermeasures to treat and/or prevent EEEV disease.

microbiology↗

bioRxiv2020

Acute exposure to sublethal doses of neonicotinoid insecticides increases heat tolerance in honey bees

The European honey bee, Apis mellifera L., is the single most valuable managed pollinator in the world. Poor colony health or unusually high colony losses of managed honey bees result from myriad stressors, which are more harmful in combination. Climate change is expected to accentuate the effects of these stressors, but the physiological and behavioral responses of honey bees to high temperatures while under simultaneous pressure of one or more stressors remains largely unknown. Here we test the hypothesis that exposure to acute, sublethal doses of neonicotinoid insecticides reduce thermal tolerance in honey bees. We administered to bees oral doses of imidacloprid and acetamiprid at 1/5, 1/20, and 1/100 of LD50 and measured their heat tolerance 4 h post-feeding, using both dynamic and static protocols. Contrary to our expectations, acute exposure to sublethal doses of both pesticides resulted in higher thermal tolerance and greater survival rates of bees. Bees that ingested the higher doses of pesticides displayed a critical thermal maximum from 2 {degrees}C to 4 {degrees}C greater than that of the control group, and a reduction in mortality from 69% to 96%. Our study suggests a resilience of honey bees to high temperatures when other stressors are present, which is consistent with studies in other insects. We discuss the implications of these results and hypothesize that this compensatory effect is likely due to induction of heat shock proteins by the insecticides, which provides temporary protection from extremely high temperatures.

physiology↗

bioRxiv2020

Lentiviral interleukin-10 gene therapy preserves fine motor circuitry and function after a cervical spinal cord injury in male and female mice

In mammals, spinal cord injuries often result in muscle paralysis through the apoptosis of lower motor neurons and denervation of neuromuscular junctions. Previous research shows that the inflammatory response to a spinal cord injury can cause additional tissue damage after the initial trauma. To modulate this inflammatory response, we delivered lentiviral anti-inflammatory interleukin-10, via loading onto an implantable biomaterial scaffold, into a left-sided hemisection at the C5 vertebra in mice. We hypothesized that improved behavioral outcomes associated with anti-inflammatory treatment are due to the sparing of fine motor circuit components. We examined behavioral recovery using a ladder beam, lower motor neuron apoptosis and muscle innervation using histology, and electromyogram recordings using intraspinal optogenetic stimulation at 2 weeks post-injury. Ladder beam analysis shows interleukin-10 treatment results in significant improvement of behavioral recovery at 2 and 12 weeks post-injury when compared to mice treated with a control virus. Histology shows interleukin-10 results in greater numbers of lower motor neurons and muscle innervation at 2 weeks post-injury. Furthermore, electromyogram recordings suggest that interleukin-10-treated animals have signal-to-noise ratios and peak-to-peak amplitudes more similar to that of uninjured controls than to that of control injured animals at 2 weeks post-injury. These data show that gene therapy using anti-inflammatory interleukin-10 can significantly reduce lower motor neuron loss, muscle denervation, and subsequent motor deficits after a spinal cord injury. Together, these results suggest that early modulation of the injury response can preserve muscle function with long-lasting benefits.

neuroscience↗

bioRxiv2019

MM-MRE: a new technique to quantify individual muscle forces during isometric tasks of the wrist using MR elastography

Non-invasive in-vivo measurement of individual muscle force is limited by the infeasibility of placing force sensing elements in series with the musculo-tendon structures. At the same time, estimating muscle forces using EMG measurements is prone to inaccuracies, as EMG is not always measurable for the complete set of muscles acting around the joints of interest. While new methods based on shear wave elastography have been recently proposed to directly characterize muscle mechanics, they can only be used to measure muscle forces in a limited set of superficial muscles. As such, they are not suitable to study the neuromuscular control of movements that require coordinated action of multiple muscles.\n\nIn this work, we present multi-muscle magnetic resonance elastography (MM-MRE), a new technique capable of quantifying individual muscle force from the complete set of muscles in the forearm, thus enabling the study of the neuromuscular control of wrist movements. MM-MRE integrates measurements of joint torque provided by an MRI-compatible instrumented handle with muscle-specific measurements of shear wave speed obtained via MRE to quantify individual muscle force using model-based estimator.\n\nA single-subject pilot experiment demonstrates the possibility of obtaining measurements from individual muscles and establishes that MM-MRE has sufficient sensitivity to detect changes in muscle mechanics following the application of isometric joint torque with self-selected intensity.

bioengineering↗

bioRxiv2017

Rare genetic variants in the endocannabinoid system genes CNR1 and DAGLA are associated with neurological phenotypes in humans

Rare genetic variants in the core endocannabinoid system genes CNR1, CNR2, DAGLA, MGLL and FAAH were identified in molecular testing data from up to 6.032 patients with a broad spectrum of neurological disorders. The variants were evaluated for association with phenotypes similar to those observed in the orthologous gene knockouts in mice. Heterozygous rare coding variants in CNR1, which encodes the type 1 cannabinoid receptor (CB1), were found to be significantly associated with pain sensitivity (especially migraine), sleep and memory disorders - alone or in combination with anxiety - compared to a set of controls without such CNR1 variants. Similarly, heterozygous rare variants in DAGLA, which encodes diacylglycerol lipase alpha, were found to be significantly associated with seizures and developmental disorders, including abnormalities of brain morphology, compared to controls. Rare variants in MGLL, FAAH and CNR2 were not associated with any neurological phenotypes in the patients tested. Diacylglycerol lipase alpha synthesizes the endocannabinoid 2-AG in the brain, which interacts with CB1 receptors. The phenotypes associated with rare CNR1 variants are reminiscent of those implicated in the theory of clinical endocannabinoid deficiency syndrome. The severe phenotypes associated with rare DAGLA variants underscore the critical role of rapid 2-AG synthesis and the endocannabinoid system in regulating neurological function and development. Mapping of the variants to the 3D structure of the type 1 cannabinoid receptor, or primary structure of diacylglycerol lipase alpha, reveals clustering of variants in certain structural regions and is consistent with impacts to function.

genetics↗