crisscrossing Science Episode 063: A Deep Dive into Sponges

In this episode, Mike Crosser (professor of physics at Linfield College) and Chad Tillberg (professor of biology at Linfield College) invite special guest Dr. Jeremy Weisz (associate professor of biology at Linfield College) into the studio to discuss sponges, including why they\u27re interesting and why they are ecologically important. It turns out sponges do more than clean your dishes

Trophic ecology of the invasive argentine ant: spatio-temporal variation in resource assimilation and isotopic enrichment

Studies of food webs often employ stable isotopic approaches to infer trophic position and interaction strength without consideration of spatio-temporal variation in resource assimilation by constituent species. Using results from laboratory diet manipulations and monthly sampling of field populations, we illustrate how nitrogen isotopes may be used to quantify spatio-temporal variation in resource assimilation in ants. First, we determined nitrogen enrichment using a controlled laboratory experiment with the invasive Argentine ant (Linepithema humile). After 12 weeks, worker δ15N values from colonies fed an animal-based diet had δ15N values that were 5.51% greater compared to colonies fed a plant-based diet. The shift in δ15N values in response to the experimental diet occurred within 10 weeks. We next reared Argentine ant colonies with or without access to honeydew-producing aphids and found that after 8 weeks workers from colonies without access to aphids had δ15N values that were 6.31% larger compared to colonies with access to honeydew. Second, we sampled field populations over a 1-year period to quantify spatio-temporal variability in isotopic ratios of L. humile and those of a common native ant (Solenopsis xyloni). Samples from free-living colonies revealed that fluctuations in δ15N were 1.6–2.4‰ for L. humile and 1.8–2.9‰ for S. xyloni. Variation was also detected among L. humile castes: time averaged means of δ15N varied from 1.2 to 2.5‰ depending on the site, with δ15N values for queens ≥ workers > brood. The estimated trophic positions of L. humile and S. xyloni were similar within a site; however, trophic position for each species differed significantly at larger spatial scales. While stable isotopes are clearly useful for examining the trophic ecology of arthropod communities, our results suggest that caution is warranted when making ecological interpretations when stable isotope collections come from single time periods or life stages

Striosome–dendron bouquets highlight a unique striatonigral circuit targeting dopamine-containing neurons

The dopamine systems of the brain powerfully influence movement and motivation. We demonstrate that striatonigral fibers originating in striosomes form highly unusual bouquet-like arborizations that target bundles of ventrally extending dopamine-containing dendrites and clusters of their parent nigral cell bodies. Retrograde tracing showed that these clustered cell bodies in turn project to the striatum as part of the classic nigrostriatal pathway. Thus, these striosome-dendron formations, here termed "striosome-dendron bouquets," likely represent subsystems with the nigro-striato-nigral loop that are affected in human disorders including Parkinson's disease. Within the bouquets, expansion microscopy resolved many individual striosomal fibers tightly intertwined with the dopamine-containing dendrites and also with afferents labeled by glutamatergic, GABAergic, and cholinergic markers and markers for astrocytic cells and fibers and connexin 43 puncta. We suggest that the striosome-dendron bouquets form specialized integrative units within the dopamine-containing nigral system. Given evidence that striosomes receive input from cortical regions related to the control of mood and motivation and that they link functionally to reinforcement and decision-making, the striosome-dendron bouquets could be critical to dopamine-related function in health and disease

A Predictive Distribution Map for the Giant Tropical Ant, Paraponera clavata

Paraponera clavata (Fabricius 1775) (Formicidae: Paraponerinae) is a widely distributed Neotropical ant whose large size has attracted the attention of numerous collectors. Working from museum specimens, a georeferenced database of collection localities was developed. This database then served as the source for computer generated predictive distribution maps. Annual rainfall was the most important variable chosen by the computer model to predict the distribution of P. clavata, both on the scale of the neotropics and at a finer scale at the northern end its distribution in Costa Rica and Nicaragua. When the model was forced to use vegetation as the first predictive variable, the Neotropical model used temperature and rainfall variance as additional variables, while the Mesoamerican model used both climatic and soils variables. Overall, the modeling suggests that P. clavata is more sensitive to abiotic factors (rainfall, temperature, soils) than to biotic factors (vegetation type) in its distribution, although this conclusion comes with the caveat that the vegetation types used in the model are quite generalized. Predictive distribution mapping holds great promise for generating more precise representations of insect distributions, thereby allowing better tests of the extent of distribution overlaps and other community relationships

BigStitcher: reconstructing high-resolution image datasets of cleared and expanded samples.

Light-sheet imaging of cleared and expanded samples creates terabyte-sized datasets that consist of many unaligned three-dimensional image tiles, which must be reconstructed before analysis. We developed the BigStitcher software to address this challenge. BigStitcher enables interactive visualization, fast and precise alignment, spatially resolved quality estimation, real-time fusion and deconvolution of dual-illumination, multitile, multiview datasets. The software also compensates for optical effects, thereby improving accuracy and enabling subsequent biological analysis

Glutamate indicators with improved activation kinetics and localization for imaging synaptic transmission

iGluSnFR variants with improved signal-to-noise ratios and targeting to postsynaptic sites have been developed, enabling the analysis of glutamatergic neurotransmission in vivo as illustrated in the mouse visual and somatosensory cortex. The fluorescent glutamate indicator iGluSnFR enables imaging of neurotransmission with genetic and molecular specificity. However, existing iGluSnFR variants exhibit low in vivo signal-to-noise ratios, saturating activation kinetics and exclusion from postsynaptic densities. Using a multiassay screen in bacteria, soluble protein and cultured neurons, we generated variants with improved signal-to-noise ratios and kinetics. We developed surface display constructs that improve iGluSnFR's nanoscopic localization to postsynapses. The resulting indicator iGluSnFR3 exhibits rapid nonsaturating activation kinetics and reports synaptic glutamate release with decreased saturation and increased specificity versus extrasynaptic signals in cultured neurons. Simultaneous imaging and electrophysiology at individual boutons in mouse visual cortex showed that iGluSnFR3 transients report single action potentials with high specificity. In vibrissal sensory cortex layer 4, we used iGluSnFR3 to characterize distinct patterns of touch-evoked feedforward input from thalamocortical boutons and both feedforward and recurrent input onto L4 cortical neuron dendritic spines

Test System Stability and Natural Variability of a Lemna Gibba L. Bioassay

BACKGROUND: In ecotoxicological and environmental studies Lemna spp. are used as test organisms due to their small size, rapid predominantly vegetative reproduction, easy handling and high sensitivity to various chemicals. However, there is not much information available concerning spatial and temporal stability of experimental set-ups used for Lemna bioassays, though this is essential for interpretation and reliability of results. We therefore investigated stability and natural variability of a Lemna gibba bioassay assessing area-related and frond number-related growth rates under controlled laboratory conditions over about one year. METHODOLOGY/PRINCIPAL FINDINGS: Lemna gibba L. was grown in beakers with Steinberg medium for one week. Area-related and frond number-related growth rates (r(area) and r(num)) were determined with a non-destructive image processing system. To assess inter-experimental stability, 35 independent experiments were performed with 10 beakers each in the course of one year. We observed changes in growth rates by a factor of two over time. These did not correlate well with temperature or relative humidity in the growth chamber. In order to assess intra-experimental stability, we analysed six systematic negative control experiments (nontoxicant tests) with 96 replicate beakers each. Evaluation showed that the chosen experimental set-up was stable and did not produce false positive results. The coefficient of variation was lower for r(area) (2.99%) than for r(num) (4.27%). CONCLUSIONS/SIGNIFICANCE: It is hypothesised that the variations in growth rates over time under controlled conditions are partly due to endogenic periodicities in Lemna gibba. The relevance of these variations for toxicity investigations should be investigated more closely. Area-related growth rate seems to be more precise as non-destructive calculation parameter than number-related growth rate. Furthermore, we propose two new validity criteria for Lemna gibba bioassays: variability of average specific and section-by-section segmented growth rate, complementary to average specific growth rate as the only validity criterion existing in guidelines for duckweed bioassays

Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies

Expansion microscopy (ExM) enables imaging of preserved specimens with nanoscale precision on diffraction-limited instead of specialized super-resolution microscopes. ExM works by physically separating fluorescent probes after anchoring them to a swellable gel. The first ExM method did not result in the retention of native proteins in the gel and relied on custom-made reagents that are not widely available. Here we describe protein retention ExM (proExM), a variant of ExM in which proteins are anchored to the swellable gel, allowing the use of conventional fluorescently labeled antibodies and streptavidin, and fluorescent proteins. We validated and demonstrated the utility of proExM for multicolor super-resolution (~70 nm) imaging of cells and mammalian tissues on conventional microscopes.United States. National Institutes of Health (1R01GM104948)United States. National Institutes of Health (1DP1NS087724)United States. National Institutes of Health ( NIH 1R01EY023173)United States. National Institutes of Health (1U01MH106011

Strategies of a parasite of the ant–Acacia mutualism

Mutualisms can be exploited by parasites—species that obtain resources from a partner but provide no services. Though the stability of mutualisms in the presence of such parasites is under intensive investigation, we have little information on life history traits that allow a species to be a successful mutualist or rather a parasite, particularly in cases where both are closely related. We studied the exploitation of Acacia myrmecophytes by the ant, Pseudomyrmex gracilis, contrasting with the mutualistic ant Pseudomyrmex ferrugineus. P. gracilis showed no host-defending behavior and had a negative effect on plant growth. By preventing the mutualist from colonization, P. gracilis imposes opportunity costs on the host plant. P. gracilis produced smaller colonies with a higher proportion of alates than did the mutualist and thus showed an “r-like” strategy. This appears to be possible because P. gracilis relies less on host-derived food resources than does the mutualist, as shown by behavioral and stable isotope studies. We discuss how this system allows the identification of strategies that characterize parasites of mutualisms