Role of P2Y12 receptor-mediated ADP signalling at microglia in the phagocytosis of neurons

Despite the prevalence of neurodegenerative disorders such as Parkinson’s and Alzheimer’s diseases, surprisingly little is known about the molecular mechanisms governing widespread neuronal loss observed in these conditions. Recent studies, however, have linked heightened microglial phagocytosis of live neurons under inflammatory stimuli such as amyloid-β and lipopolysaccharide (LPS) to this loss. The ADP-specific microglial receptor P2Y12 has been implicated in long-range communication between neurons and microglia, as well as in microglia chemotaxis preceding phagocytosis, but the direct role of this receptor in phagocytosis of live neurons has not been addressed. In addition, P2Y12 antagonists have already been established in the clinic as treatments for thrombosis, and whether these inhibitors could further serve a neuroprotective purpose by preventing phagocytosis was speculated. Upon examining the effect of P2Y12 inhibition using the competitive antagonist PSB0739, and P2Y12 genetic knockdown on phagocytosis of neuroblast-like, naïve PC12 targets by LPS-activated BV-2 microglia through a combination of flow cytometry and microscopy analyses, I show that P2Y12 perturbation does not affect microglial phagocytosis of neuronal targets under LPS stress. However, P2Y12 disruption led to significant decreases in phagocytosis of bead and neuronal targets under unstimulated conditions, which suggested that P2Y12 plays an important role in the phagocytic mechanisms of resting microglia. Additionally, fluorescent calcium response assays yielded significant decreases in acute calcium response to ADP in P2Y12-inhibited and knockdown cells, confirming that P2Y12 plays a major role in calcium mobilization due to ADP treatment in BV-2 microglia. Furthermore, examination of the role of ADP on the regulation of P2Y12 mRNA expression using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) led to the finding that P2Y12 expression is reduced in response to high concentrations of ADP. These findings highlight the novel ideas that 1) the P2Y12 receptor has functional roles in phagocytosis of basal microglia besides cell migration, and 2) persistent imbalance of ADP in the extracellular environment affects P2Y12 expression, which may underlie inflammatory stimuli-induced P2Y12 downregulation in the transition from resting to highly phagocytic, disease-associated microglia

Updating existing emotional memories involves the frontopolar/orbitofrontal cortex in ways that acquiring new emotional memories does not

In life, we must often learn new associations to people, places, or things we already know. The current fMRI study investigated the neural mechanisms underlying emotional memory updating. Nineteen participants first viewed negative and neutral pictures and learned associations between those pictures and other neutral stimuli, such as neutral objects and encoding tasks. This initial learning phase was followed by a memory updating phase, during which participants learned picture-location associations for old pictures (i.e., pictures previously associated with other neutral stimuli) and new pictures (i.e., pictures not seen in the first phase). There was greater frontopolar/orbito-frontal (OFC) activity when people learned picture–location associations for old negative pictures than for new negative pictures, but frontopolar OFC activity did not significantly differ during learning locations of old versus new neutral pictures. In addition, frontopolar activity was more negatively correlated with the amygdala when participants learned picture–location associations for old negative pictures than for new negative or old neutral pictures. Past studies revealed that the frontopolar OFC allows for updating the affective values of stimuli in reversal learning or extinction of conditioning [e.g., Izquierdo, A., & Murray, E. A. Opposing effects of amygdala and orbital PFC lesions on the extinction of instrumental responding in macaque monkeys. European Journal of Neuroscience, 22, 2341–2346, 2005]; our findings suggest that it plays a more general role in updating associations to emotional stimuli

First Observations of a "Fast Beam-Ion Instability"

We report the results of experiments on a "fast beam-ion instability" at the Advanced Light Source (ALS). This ion instability, which can arise even where the ions are not trapped over multiple beam passages, will likely be important for many future accelerators. In our experiments, we filled the ALS storage ring with helium gas, raising the pressure approximately two orders of magnitude above the nominal value. With gaps in the bunch train large enough to avoid conventional (multi-turn) ion trapping, we observed a factor of 2-3 increase in the vertical beam size along with coherent beam oscillations. The single-pass nature of this instability was demonstrated

Identification of Potential Calorie Restriction-Mimicking Yeast Mutants with Increased Mitochondrial Respiratory Chain and Nitric Oxide Levels

Calorie restriction (CR) induces a metabolic shift towards mitochondrial respiration; however, molecular mechanisms underlying CR remain unclear. Recent studies suggest that CR-induced mitochondrial activity is associated with nitric oxide (NO) production. To understand the role of mitochondria in CR, we identify and study Saccharomyces cerevisiae mutants with increased NO levels as potential CR mimics. Analysis of the top 17 mutants demonstrates a correlation between increased NO, mitochondrial respiration, and longevity. Interestingly, treating yeast with NO donors such as GSNO (S-nitrosoglutathione) is sufficient to partially mimic CR to extend lifespan. CR-increased NO is largely dependent on mitochondrial electron transport and cytochrome c oxidase (COX). Although COX normally produces NO under hypoxic conditions, CR-treated yeast cells are able to produce NO under normoxic conditions. Our results suggest that CR may derepress some hypoxic genes for mitochondrial proteins that function to promote the production of NO and the extension of lifespan

Measurement with beam of the deflecting higher order modes in the TTF superconducting cavities

This paper reports on recent beam measurements of higher order modes in the TESLA Test Facility (TTF) accelerating modules. Using bunch trains of about 0.5 ms with 54MHz bunch repetition and up to 90% modulated intensity, transverse higher order modes are resonantly excited when the beam is offset and their frequency on resonance with the modulation frequency. With this method, the trapped modes can be excited and their counteraction on the beam observed on a wide-band BPM downstream of the module. Scanning the modulation frequency from 0 to 27MHz allows a systematic investigation of all possible dangerous modes in the modules

Identification of the first ATRIP-deficient patient and novel mutations in ATR define a clinical spectrum for ATR-ATRIP Seckel Syndrome

A homozygous mutational change in the Ataxia-Telangiectasia and RAD3 related (ATR) gene was previously reported in two related families displaying Seckel Syndrome (SS). Here, we provide the first identification of a Seckel Syndrome patient with mutations in ATRIP, the gene encoding ATR-Interacting Protein (ATRIP), the partner protein of ATR required for ATR stability and recruitment to the site of DNA damage. The patient has compound heterozygous mutations in ATRIP resulting in reduced ATRIP and ATR expression. A nonsense mutational change in one ATRIP allele results in a C-terminal truncated protein, which impairs ATR-ATRIP interaction; the other allele is abnormally spliced. We additionally describe two further unrelated patients native to the UK with the same novel, heterozygous mutations in ATR, which cause dramatically reduced ATR expression. All patient-derived cells showed defective DNA damage responses that can be attributed to impaired ATR-ATRIP function. Seckel Syndrome is characterised by microcephaly and growth delay, features also displayed by several related disorders including Majewski (microcephalic) osteodysplastic primordial dwarfism (MOPD) type II and Meier-Gorlin Syndrome (MGS). The identification of an ATRIP-deficient patient provides a novel genetic defect for Seckel Syndrome. Coupled with the identification of further ATR-deficient patients, our findings allow a spectrum of clinical features that can be ascribed to the ATR-ATRIP deficient sub-class of Seckel Syndrome. ATR-ATRIP patients are characterised by extremely severe microcephaly and growth delay, microtia (small ears), micrognathia (small and receding chin), and dental crowding. While aberrant bone development was mild in the original ATR-SS patient, some of the patients described here display skeletal abnormalities including, in one patient, small patellae, a feature characteristically observed in Meier-Gorlin Syndrome. Collectively, our analysis exposes an overlapping clinical manifestation between the disorders but allows an expanded spectrum of clinical features for ATR-ATRIP Seckel Syndrome to be define

Elicitin-responsive lectin-like receptor kinase genes in BY-2 cells

The inhibition of elicitor-induced plant defense responses by the protein kinase inhibitors K252a and staurosporine indicates that defense responses require protein phosphorylation. We isolated a cDNA clone encoding Nicotiana tabacum lectin-like receptor protein kinase 1 ( NtlecRK1), an elicitor-responsive gene; in tobacco bright yellow ( BY-2) cells by a differential display method. NtlecRK forms a gene family with at least three members in tobacco. All three NtlecRK genes potentially encode the N-terminal legume lectin domain, transmembrane domain and C-terminal Ser/Thr-type protein kinase domain. Green fluorescent protein ( GFP) fusion showed that the NtlecRK1 protein was located on the plasma membrane. In addition, NtlecRK1 and 3 were responsive to INF1 elicitin and the bacterial elicitor harpin. These results indicate that NtlecRKs are membrane-located protein kinases that are induced during defense responses in BY-2 cells.</p

Beam and SKS spectrometers at the K1.8 beam line

High-resolution spectrometers for both incident beams and scattered particles have been constructed at the K1.8 beam line of the Hadron Experimental Facility at J-PARC. A point-to-point optics is realized between the entrance and exit of QQDQQ magnets for the beam spectrometer. Fine-pitch wire chamber trackers and hodoscope counters are installed in the beam spectrometer to accept a high rate beam up to 107 Hz. The superconducting kaon spectrometer for scattered particles was transferred from KEK with modifications to the cryogenic system and detectors. A missing-mass resolution of 1.9 ± 0.1 MeV/c2 (FWHM) was achieved for the ∑ peaks of (π±, K+) reactions on a proton target in the first physics run of E19 in 2010

Innovations for Sustainability: Pathways to an efficient and post-pandemic future

3rd Report prepared by The World in 2050 initiativ