Immunocytochemistry by Electron Spectroscopic Imaging Using Well Defined Boronated Monovalent Antibody Fragments

Contributing to the rapidly developing field of immunoelectron microscopy a new kind of markers has been created. The element boron, incorporated as very stable carborane clusters into different kinds of peptides, served as a marker detectable by electron spectroscopic imaging (ESI) - an electron microscopic technique with high-resolution potential. Covalently linked immunoreagents conspicuous by the small size of both antigen recognizing part and marker moiety are accessible by using peptide concepts for label construction and their conjugation with Fab\u27 fragments. Due to a specific labeling of the free thiol groups of the Fab\u27 fragments, the antigen binding capacity was not affected by the attachment of the markers and the resulting immunoprobes exhibited an elongated shape with the antigen combining site and the label located at opposite ends. The labeling densities observed with these reagents were found to be significantly higher than those obtained by using conventional colloidal gold methods. Combined with digital image processing and analysis systems, boron-based ESI proved to be a powerful approach in ultrastructural immunocytochemistry employing pre-and post-embedding methods

Regulation of Drosophila Brain Wiring by Neuropil Interactions via a Slit-Robo-RPTP Signaling Complex

The axonal wiring molecule Slit and its Round-About (Robo) receptors are conserved regulators of nerve cord patterning. Robo receptors also contribute to wiring brain circuits. Whether molecular mechanisms regulating these signals are modified to fit more complex brain wiring processes is unclear. We investigated the role of Slit and Robo receptors in wiring Drosophila higher-order brain circuits and identified differences in the cellular and molecular mechanisms of Robo/Slit function. First, we find that signaling by Robo receptors in the brain is regulated by the Receptor Protein Tyrosine Phosphatase RPTP69d. RPTP69d increases membrane availability of Robo3 without affecting its phosphorylation state. Second, we detect no midline localization of Slit during brain development. Instead, Slit is enriched in the mushroom body, a neuronal structure covering large areas of the brain. Thus, a divergent molecular mechanism regulates neuronal circuit wiring in the Drosophila brain, partly in response to signals from the mushroom body

A monocarboxylate transporter rescues frontotemporal dementia and Alzheimer's disease models

Brains are highly metabolically active organs, consuming 20% of a person's energy at resting state. A decline in glucose metabolism is a common feature across a number of neurodegenerative diseases. Another common feature is the progressive accumulation of insoluble protein deposits, it's unclear if the two are linked. Glucose metabolism in the brain is highly coupled between neurons and glia, with glucose taken up by glia and metabolised to lactate, which is then shuttled via transporters to neurons, where it is converted back to pyruvate and fed into the TCA cycle for ATP production. Monocarboxylates are also involved in signalling, and play broad ranging roles in brain homeostasis and metabolic reprogramming. However, the role of monocarboxylates in dementia has not been tested. Here, we find that increasing pyruvate import in Drosophila neurons by over-expression of the transporter bumpel, leads to a rescue of lifespan and behavioural phenotypes in fly models of both frontotemporal dementia and Alzheimer's disease. The rescue is linked to a clearance of late stage autolysosomes, leading to degradation of toxic peptides associated with disease. We propose upregulation of pyruvate import into neurons as potentially a broad-scope therapeutic approach to increase neuronal autophagy, which could be beneficial for multiple dementias

A ∼\sim15 kpc outflow cone piercing through the halo of the blue compact metal-poor galaxy SBS0335-052

Context: Outflows from low-mass star-forming galaxies are a fundamental ingredient for models of galaxy evolution and cosmology. Aims: The onset of kpc-scale ionised filaments in the halo of the metal-poor compact dwarf SBS 0335-052E was previously not linked to an outflow. We here we investigate whether these filaments provide evidence for an outflow. Methods: We obtained new VLT/MUSE WFM and deep NRAO/VLA B-configuration 21cm data of the galaxy. The MUSE data provide morphology, kinematics, and emission line ratios H$\beta$/H$\alpha$ and [\ion{O}{iii}]$\lambda5007$/H$\alpha$ of the low surface-brightness filaments, while the VLA data deliver morphology and kinematics of the neutral gas in and around the system. Both datasets are used in concert for comparisons between the ionised and the neutral phase. Results: We report the prolongation of a lacy filamentary ionised structure up to a projected distance of 16 kpc at $\mathrm{SB}_\mathrm{H\alpha} = 1.5\times10^{-18}$erg s$^{-1}$ cm$^{-2}$arcsec$^{-2}$. The filaments exhibit unusual low H$\alpha$/H$\beta \approx 2.4$ and low [\ion{O}{iii}]/H$\alpha \sim 0.4 - 0.6$ typical of diffuse ionised gas. They are spectrally narrow ($\sim 20$ km s$^{-1}$) and exhibit no velocity sub-structure. The filaments extend outwards of the elongated \ion{H}{I} halo. On small scales the $N_\mathrm{HI}$ peak is offset from the main star-forming sites. Morphology and kinematics of \ion{H}{I} and \ion{H}{II} reveal how star-formation driven feedback interacts differently with the ionised and the neutral phase. Conclusions: We reason that the filaments are a large scale manifestation of star-formation driven feedback, namely limb-brightened edges of a giant outflow cone that protrudes through the halo of this gas-rich system. A simple toy model of such a conical-structure is found to be commensurable with the observations.Comment: Accepted version in A&A after language editing. 22 pages, 24 figure

Late Quaternary paleoenvironmental records from the Chatanika River valley near Fairbanks (Alaska)

© 2016 Elsevier LtdPerennially-frozen deposits are considered as excellent paleoenvironmental archives similar to lacustrine, deep marine, and glacier records because of the long-term and good preservation of fossil records under stable permafrost conditions. A permafrost tunnel in the Vault Creek Valley (Chatanika River Valley, near Fairbanks) exposes a sequence of frozen deposits and ground ice that provides a comprehensive set of proxies to reconstruct the late Quaternary environmental history of Interior Alaska. The multi-proxy approach includes different dating techniques (radiocarbon-accelerator mass spectrometry [AMS 14C], optically stimulated luminescence [OSL], thorium/uranium radioisotope disequilibria [230Th/U]), as well as methods of sedimentology, paleoecology, hydrochemistry, and stable isotope geochemistry of ground ice. The studied sequence consists of 36-m-thick late Quaternary deposits above schistose bedrock. Main portions of the sequence accumulated during the early and middle Wisconsin periods. The lowermost unit A consists of about 9-m-thick ice-bonded fluvial gravels with sand and peat lenses. A late Sangamon (MIS 5a) age of unit A is assumed. Spruce forest with birch, larch, and some shrubby alder dominated the vegetation. High presence of Sphagnum spores and Cyperaceae pollen points to mires in the Vault Creek Valley. The overlying unit B consists of 10-m-thick alternating fluvial gravels, loess-like silt, and sand layers, penetrated by small ice wedges. OSL dates support a stadial early Wisconsin (MIS 4) age of unit B. Pollen and plant macrofossil data point to spruce forests with some birch interspersed with wetlands around the site. The following unit C is composed of 15-m-thick ice-rich loess-like and organic-rich silt with fossil bones and large ice wedges. Unit C formed during the interstadial mid-Wisconsin (MIS 3) and stadial late Wisconsin (MIS 2) as indicated by radiocarbon ages. Post-depositional slope processes significantly deformed both, ground ice and sediments of unit C. Pollen data show that spruce forests and wetlands dominated the area. The macrofossil remains of Picea, Larix, and Alnus incana ssp. tenuifolia also prove the existence of boreal coniferous forests during the mid-Wisconsin interstadial, which were replaced by treeless tundra-steppe vegetation during the late Wisconsin stadial. Unit C is discordantly overlain by the 2-m-thick late Holocene deposits of unit D. The pollen record of unit D indicates boreal forest vegetation similar to the modern one. The permafrost record from the Vault Creek tunnel reflects more than 90 ka of periglacial landscape dynamics triggered by fluvial and eolian accumulation, and formation of ice-wedge polygons and post-depositional deformation by slope processes. The record represents a typical Wisconsin valley-bottom facies in Central Alaska

DLGS97/SAP97 is developmentally upregulated and is required for complex adult behaviors and synapse morphology and function

The synaptic membrane-associated guanylate kinase (MAGUK) scaffolding protein family is thought to play key roles in synapse assembly and synaptic plasticity. Evidence supporting these roles in vivo is scarce, as a consequence of gene redundancy in mammals. The genome of Drosophila contains only one MAGUK gene, discs large (dlg), from which two major proteins originate: DLGA [PSD95 (postsynaptic density 95)-like] and DLGS97 [SAP97 (synapse-associated protein)-like]. These differ only by the inclusion in DLGS97 of an L27 domain, important for the formation of supramolecular assemblies. Known dlg mutations affect both forms and are lethal at larval stages attributable to tumoral overgrowth of epithelia. We generated independent null mutations for each, dlgA and dlgS97. These allowed unveiling of a shift in expression during the development of the nervous system: predominant expression of DLGA in the embryo, balanced expression of both during larval stages, and almost exclusive DLGS97 expression in the adult brain. Loss of embryonic DLGS97 does not alter the development of the nervous system. At larval stages, DLGA and DLGS97 fulfill both unique and partially redundant functions in the neuromuscular junction. Contrary to dlg and dlgA mutants, dlgS97 mutants are viable to adulthood, but they exhibit marked alterations in complex behaviors such as phototaxis, circadian activity, and courtship, whereas simpler behaviors like locomotion and odor and light perception are spared. We propose that the increased repertoire of associations of a synaptic scaffold protein given by an additional domain of protein-protein interaction underlies its ability to integrate molecular networks required for complex functions in adult synapses

Physical properties of Centaur (60558) 174P/Echeclus from stellar occultations

The Centaur (60558) Echeclus was discovered on March 03, 2000, orbiting between the orbits of Jupiter and Uranus. After exhibiting frequent outbursts, it also received a comet designation, 174P. If the ejected material can be a source of debris to form additional structures, studying the surroundings of an active body like Echeclus can provide clues about the formation scenarios of rings, jets, or dusty shells around small bodies. Stellar occultation is a handy technique for this kind of investigation, as it can, from Earth-based observations, detect small structures with low opacity around these objects. Stellar occultation by Echeclus was predicted and observed in 2019, 2020, and 2021. We obtain upper detection limits of rings with widths larger than 0.5 km and optical depth of $\tau$ = 0.02. These values are smaller than those of Chariklo's main ring; in other words, a Chariklo-like ring would have been detected. The occultation observed in 2020 provided two positive chords used to derive the triaxial dimensions of Echeclus based on a 3D model and pole orientation available in the literature. We obtained $a = 37.0\pm0.6$ km, $b = 28.4 \pm 0.5$ km, and $c= 24.9 \pm 0.4$ km, resulting in an area-equivalent radius of $30.0 \pm 0.5$ km. Using the projected limb at the occultation epoch and the available absolute magnitude ($\rm{H}_{\rm{v}} = 9.971 \pm 0.031$), we calculate an albedo of $p_{\rm{v}} = 0.050 \pm 0.003$. Constraints on the object's density and internal friction are also proposed.Comment: Corrected and typeset versio