Dissecting the Extracellular Complexity of Neuromuscular Junction Organizers

Synapse formation is a very elaborate process dependent upon accurate coordination of pre and post-synaptic specialization, requiring multiple steps and a variety of receptors and signaling molecules. Due to its relative structural simplicity and the ease in manipulation and observation, the neuromuscular synapse or neuromuscular junction (NMJ)—the connection between motor neurons and skeletal muscle—represents the archetype junction system for studying synapse formation and conservation. This junction is essential for survival, as it controls our ability to move and breath. NMJ formation requires coordinated interactions between motor neurons and muscle fibers, which ultimately result in the formation of a highly specialized post-synaptic architecture and a highly differentiated nerve terminal. Furthermore, to ensure a fast and reliable synaptic transmission following neurotransmitter release, ligand-gated channels (acetylcholine receptors, AChRs) are clustered on the post-synaptic muscle cell at high concentrations in sites opposite the presynaptic active zone, supporting a direct role for nerves in the organization of the post-synaptic membrane architecture. This organized clustering process, essential for NMJ formation and for life, relies on key signaling molecules and receptors and is regulated by soluble extracellular molecules localized within the synaptic cleft. Notably, several mutations as well as auto-antibodies against components of these signaling complexes have been related to neuromuscular disorders. The recent years have witnessed strong progress in the understanding of molecular identities, architectures, and functions of NMJ macromolecules. Among these, prominent roles have been proposed for neural variants of the proteoglycan agrin, its receptor at NMJs composed of the lipoprotein receptor-related protein 4 (LRP4) and the muscle-specific kinase (MuSK), as well as the regulatory soluble synapse-specific protease Neurotrypsin. In this review we summarize the current state of the art regarding molecular structures and (agrin-dependent) canonical, as well as (agrin-independent) non-canonical, MuSK signaling mechanisms that underscore the formation of neuromuscular junctions, with the aim of providing a broad perspective to further stimulate molecular, cellular and tissue biology investigations on this fundamental intercellular contact

Direct experimental observation of nonclassicality in ensembles of single photon emitters

In this work we experimentally demonstrate for the first time a recently proposed criterion adressed to detect nonclassical behavior in the fluorescence emission of ensembles of single-photon emitters. In particular, we apply the method to study clusters of NV centres in diamond observed via single-photon-sensitive confocal microscopy. Theoretical considerations on the behavior of the parameter at any arbitrary order in presence of poissonian noise are presented and, finally, the opportunity of detecting manifold coincidences is discussed

Native NIR-emitting single colour centres in CVD diamond

Single-photon sources are a fundamental element for developing quantum technologies, and sources based on colour centres in diamonds are among the most promising candidates. The well-known NV centres are characterized by several limitations, thus few other defects have recently been considered. In the present work, we characterize in detail native efficient single colour centres emitting in the near infra-red in both standard IIa single-crystal and electronic-grade polycrystalline commercial CVD diamond samples. In the former case, a high-temperature annealing process in vacuum is necessary to induce the formation/activation of luminescent centres with good emission properties, while in the latter case the annealing process has marginal beneficial effects on the number and performances of native centres in commercially available samples. Although displaying significant variability in several photo physical properties (emission wavelength, emission rate instabilities, saturation behaviours), these centres generally display appealing photophysical properties for applications as single photon sources: short lifetimes, high emission rates and strongly polarized light. The native centres are tentatively attributed to impurities incorporated in the diamond crystal during the CVD growth of high-quality type IIa samples, and offer promising perspectives in diamond-based photonics.Comment: 27 pages, 10 figures. Submitted to "New Journal of Phsyics", NJP-100003.R

Ефективність симбіотичної азотфіксації в агроценозах України

Проведено порівняльну оцінку ефективності виробничих і перспективних штамів бульбочкових бактерій колекції Південної дослідної станції ІСГМ УААН у симбіозі з сучасними сортами бобових культур. Визначені високоефективні комбінації “сорти – штами”. Показано доцільність передпосівної інокуляції насіння бобових культур на фоні ґрунтової популяції ризобій.Проведена сравнительная оценка эффективности производственных и перспективных штаммов клубеньковых бактерий коллекции Южной опытной станции ИСХМ УААН в симбиозе с современными сортами бобовых культур. Определены высокоэффективные комбинации “сорта – штаммы”. Показана целесообразность предпосевной инокуляции семян бобовых культур на фоне почвенной популяции ризобий.The comparative estimation of rhisobia strains (from the collection of the Southern Experimental Station of Institute of Agricultural Microbiology UAAS) efficiency in symbiosis with modern legume cultivars was done. Highly effective complemented „cultivars – strains” combinations have been determined. It was shown the expediency of legume seeds pre-sowing treatment on background of soil rhisobia population

Nanodiamonds-induced effects on neuronal firing of mouse hippocampal microcircuits

Fluorescent nanodiamonds (FND) are carbon-based nanomaterials that can efficiently incorporate optically active photoluminescent centers such as the nitrogen-vacancy complex, thus making them promising candidates as optical biolabels and drug-delivery agents. FNDs exhibit bright fluorescence without photobleaching combined with high uptake rate and low cytotoxicity. Focusing on FNDs interference with neuronal function, here we examined their effect on cultured hippocampal neurons, monitoring the whole network development as well as the electrophysiological properties of single neurons. We observed that FNDs drastically decreased the frequency of inhibitory (from 1.81 Hz to 0.86 Hz) and excitatory (from 1.61 Hz to 0.68 Hz) miniature postsynaptic currents, and consistently reduced action potential (AP) firing frequency (by 36%), as measured by microelectrode arrays. On the contrary, bursts synchronization was preserved, as well as the amplitude of spontaneous inhibitory and excitatory events. Current-clamp recordings revealed that the ratio of neurons responding with AP trains of high-frequency (fast-spiking) versus neurons responding with trains of low-frequency (slow-spiking) was unaltered, suggesting that FNDs exerted a comparable action on neuronal subpopulations. At the single cell level, rapid onset of the somatic AP ("kink") was drastically reduced in FND-treated neurons, suggesting a reduced contribution of axonal and dendritic components while preserving neuronal excitability.Comment: 34 pages, 9 figure

Photo-physical properties of He-related color centers in diamond

Diamond is a promising platform for the development of technological applications in quantum optics and photonics. The quest for color centers with optimal photo-physical properties has led in recent years to the search for novel impurity-related defects in this material. Here, we report on a systematic investigation of the photo-physical properties of two He-related (HR) emission lines at 535 nm and 560 nm created in three different diamond substrates upon implantation with 1.3 MeV He+ ions and subsequent annealing. The spectral features of the HR centers were studied in an "optical grade" diamond substrate as a function of several physical parameters, namely the measurement temperature, the excitation wavelength and the intensity of external electric fields. The emission lifetimes of the 535 nm and 560 nm lines were also measured by means of time-gated photoluminescence measurements, yielding characteristic decay times of (29 +- 5) ns and (106 +- 10) ns, respectively. The Stark shifting of the HR centers under the application of an external electrical field was observed in a CVD diamond film equipped with buried graphitic electrodes, suggesting a lack of inversion symmetry in the defects' structure. Furthermore, the photoluminescence mapping under 405 nm excitation of a "detector grade" diamond sample implanted at a 1x1010 cm-2 He+ ion fluence enabled to identify the spectral features of both the HR emission lines from the same localized optical spots. The reported results provide a first insight towards the understanding of the structure of He-related defects in diamond and their possible utilization in practical applicationsComment: 9 pages, 3 figure

A 3-dimensional interdigitated electrode geometry for the enhancement of charge collection efficiency in diamond detectors

In this work, a single crystal CVD diamond film with a novel three-dimensional (3D) interdigitated electrode geometry has been fabricated with the reactive ion etching (RIE) technique in order to increase the charge collection efficiency (CCE) with respect to that obtained by standard superficial electrodes. The geometrical arrangement of the electric field lines due to the 3D patterning of the electrodes results in a shorter travel path for the excess charge carriers, thus contributing to a more efficient charge collection mechanism. The CCE of the device was mapped by means of the ion beam induced charge (IBIC) technique. A 1MeV proton microbeam was raster-scanned over the active area of the diamond detector under different bias voltage conditions, enabling to probe the charge transport properties of the detector up to a depth of 8 ìm below the sample surface. The experimental results, supported by the numerical simulations, show a significant improvement in the 3D detector performance (i.e. CCE, energy resolution, extension of the active area) if compared with the results obtained by standard surface metallic electrodes