Sex-specific transcriptional and proteomic signatures in schizophrenia

It has remained unclear why schizophrenia typically manifests after adolescence and which neurobiological mechanisms are underlying the cascade leading to the actual onset of the illness. Here we show that the use of induced pluripotent stem cell-derived neurons of monozygotic twins from pairs discordant for schizophrenia enhances disease-specific signal by minimizing genetic heterogeneity. In proteomic and pathway analyses, clinical illness is associated especially with altered glycosaminoglycan, GABAergic synapse, sialylation, and purine metabolism pathways. Although only 12% of all 19,462 genes are expressed differentially between healthy males and females, up to 61% of the illness-related genes are sex specific. These results on sex-specific genes are replicated in another dataset. This implies that the pathophysiology differs between males and females, and may explain why symptoms appear after adolescence when the expression of many sex-specific genes change, and suggests the need for sex-specific treatments.Peer reviewe

European contribution to the study of ROS: A summary of the findings and prospects for the future from the COST action BM1203 (EU-ROS).

The European Cooperation in Science and Technology (COST) provides an ideal framework to establish multi-disciplinary research networks. COST Action BM1203 (EU-ROS) represents a consortium of researchers from different disciplines who are dedicated to providing new insights and tools for better understanding redox biology and medicine and, in the long run, to finding new therapeutic strategies to target dysregulated redox processes in various diseases. This report highlights the major achievements of EU-ROS as well as research updates and new perspectives arising from its members. The EU-ROS consortium comprised more than 140 active members who worked together for four years on the topics briefly described below. The formation of reactive oxygen and nitrogen species (RONS) is an established hallmark of our aerobic environment and metabolism but RONS also act as messengers via redox regulation of essential cellular processes. The fact that many diseases have been found to be associated with oxidative stress established the theory of oxidative stress as a trigger of diseases that can be corrected by antioxidant therapy. However, while experimental studies support this thesis, clinical studies still generate controversial results, due to complex pathophysiology of oxidative stress in humans. For future improvement of antioxidant therapy and better understanding of redox-associated disease progression detailed knowledge on the sources and targets of RONS formation and discrimination of their detrimental or beneficial roles is required. In order to advance this important area of biology and medicine, highly synergistic approaches combining a variety of diverse and contrasting disciplines are needed.The EU-ROS consortium (COST Action BM1203) was supported by the European Cooperation in Science and Technology (COST). The present overview represents the final Action dissemination summarizing the major achievements of COST Action BM1203 (EU-ROS) as well as research news and personal views of its members. Some authors were also supported by COST Actions BM1005 (ENOG) and BM1307 (PROTEOSTASIS), as well as funding from the European Commission FP7 and H2020 programmes, and several national funding agencies

GluN2A subunit-containing NMDA receptors are the preferential neuronal targets of homocysteine

Homocysteine (HCY) is an endogenous redox active amino acid, best known as contributor to various neurodegenerative disorders. Although it is known that HCY can activate NMDA receptors (NMDARs), the mechanisms of its action on receptors composed of different NMDA receptor subunits remains almost unknown. In this study, using imaging and patch clamp technique in cultured cortical neurons and heterologous expression in HEK293T cells we tested the agonist activity of HCY on NMDARs composed of GluN1 and GluN2A subunits (GluN1/2A receptors) and GluN1 and GluN2B subunits (GluN1/2B receptors). We demonstrate that the time courses of Ca2+ transients and membrane currents activated by HCY and NMDA in cortical neurons are drastically different. Application of HCY to cortical neurons induced responses, which in contrast to currents induced by NMDA (both in the presence of glycine) considerably decreased to steady state of small amplitude. In contrast to NMDA, HCY-activated currents at steady state were resistant to the selective GluN2B subunit inhibitor ifenprodil. In calcium-free external solution the decrease of NMDA evoked currents was abolished, suggesting the Ca2+-dependent NMDAR desensitization. Under these conditions HCY evoked currents still declined almost to the baseline suggesting Ca2+-independent desensitization. In HEK293T cells HCY activated NMDARs of GluN1/2A and GluN1/2B subunit compositions with EC50s of 9.7 ± 1.8 μM and 61.8 ± 8.9 μM, respectively. Recombinant GluN1/2A receptors, however, did not desensitize by HCY, whereas GluN1/2B receptors were almost fully desensitized by HCY. Thus, HCY is a high affinity agonist of NMDARs preferring the GluN1/2A subunit composition. Our data suggest that HCY induced native NMDAR currents in neurons are mainly mediated by the synaptic type GluN1/2A NMDARs. This implies that in hyperhomocysteinemia, a disorder with enlarged level of HCY in plasma, HCY may persistently contribute to postsynaptic responses mediated by GluN2A-containing NMDA receptors. On the other hand, HCY toxicity may be limited by desensitization typical for HCY-induced activation of GluN2B-containing extrasynaptic receptors. Our findings, therefore, provide an evidence for the physiological relevance of endogenous HCY, which may represent an effective endogenous modulator of the central excitatory neurotransmission

Functional Properties of Human NMDA Receptors Associated with Epilepsy-Related Mutations of GluN2A Subunit

International audienceGenetic variants of the glutamate activated N-methyl-D-aspartate (NMDA) receptor (NMDAR) subunit GluN2A are associated with the hyperexcitable states manifested by epileptic seizures and interictal discharges in patients with disorders of the epilepsy-aphasia spectrum (EAS). The variants found in sporadic cases and families are of different types and include microdeletions encompassing the corresponding GRIN2A gene as well as nonsense, splice-site and missenseGRIN2A defects. They are located at different functional domains of GluN2A and no clear genotype-phenotype correlation has emerged yet. Moreover, GluN2A variants may be associated with phenotypic pleiotropy. Deciphering the consequences of pathogenic GRIN2A variants would surely help in better understanding of the underlying mechanisms. This emphasizes the need for functional studies to unravel the basic functional properties of each specific NMDAR variant. In the present study, we have used patch-clamp recordings to evaluate kinetic changes of mutant NMDARs reconstituted after co-transfection of cultured cells with the appropriate expression vectors. Three previously identified missense variants found in patients or families with disorders of the EAS and situated in the N-terminal domain (p.Ile184Ser) or in the ligand-binding domain (p.Arg518His and p.Ala716Thr) of GluN2A were studied in both the homozygous and heterozygous conditions. Relative surface expression and current amplitude were significantly reduced for NMDARs composed of mutant p.Ile184Ser and p.Arg518His, but not p.Ala716His, as compared with wild-type (WT) NMDARs. Amplitude of whole-cell currents was still drastically decreased when WT and mutant p.Arg518His-GluN2A subunits were co-expressed, suggesting a dominant-negative mechanism. Activation times were significantly decreased in both homozygous and heterozygous conditions for the two p.Ile184Ser and p.Arg518His variants, but not for p.Ala716His. Deactivation also significantly increased for p.Ile184Ser variant in the homozygous but not the heterozygousstate while it was increased for p.Arg518His in both states. Our data indicate that p.Ile184Ser and p.Arg518His GluN2A variants both impacted on NMDAR function, albeit differently, whereas p.Ala716His did not significantly influence NMDAR kinetics, hence partly questioning its direct and strong pathogenic role. This study brings new insights into the functional impact that GRIN2A variants might have on NMDAR kinetics, and provides a mechanistic explanation for the neurological manifestations seen in the corresponding human spectrum of disorders

Variability of microcirculation detected by blood pulsation imaging.

The non-invasive assessment of blood flow is invaluable for the diagnostic and monitoring treatment of numerous vascular and neurological diseases. We developed a non-invasive and non-contact method of blood pulsation imaging capable of visualizing and monitoring of the two-dimensional distribution of two key parameters of peripheral blood flow: the blood pulsation amplitude and blood pulsation phase. The method is based on the photoplethysmographic imaging in the reflection mode. In contrast with previous imaging systems we use new algorithm for data processing which allows two dimensional mapping of blood pulsations in large object's areas after every cardiac cycle. In our study we carried out the occlusion test of the arm and found (i) the extensive variability of 2D-distribution of blood pulsation amplitude from one cardiac cycle to another, and (ii) existence of the adjacent spots to which the blood is asynchronously supplied. These observations show that the method can be used for studying of the multicomponent regulation of peripheral blood circulation. The proposed technique is technologically simple and cost-effective, which makes it applicable for monitoring the peripheral microcirculation in clinical settings for example, in diagnostics or testing the efficiency of new medicines

Origin of Infrared Light Modulation in Reflectance-Mode Photoplethysmography.

We recently pointed out the important role of dermis deformation by pulsating arterial pressure in the formation of a photoplethysmographic signal at green light. The aim of this study was to explore the role of this novel finding in near-infrared (NIR) light. A light-emitting diode (LED)-based imaging photoplethysmography (IPPG) system was used to detect spatial distribution of blood pulsations under frame-to-frame switching green and NIR illumination in the palms of 34 healthy individuals. We observed a significant increase of light-intensity modulation at the heartbeat frequency for both illuminating wavelengths after a palm was contacted with a glass plate. Strong positive correlation between data measured at green and NIR light was found, suggesting that the same signal was read independently from the depth of penetration. Analysis of the data shows that an essential part of remitted NIR light is modulated in time as a result of elastic deformations of dermis caused by variable blood pressure in the arteries. Our observations suggest that in contrast with the classical model, photoplethysmographic waveform originates from the modulation of the density of capillaries caused by the variable pressure applied to the skin from large blood vessels. Particularly, beat-to-beat transmural pressure in arteries compresses/decompresses the dermis and deforms its connective-tissue components, thus affecting the distance between the capillaries, which results in the modulation of absorption and scattering coefficients of both green and NIR light. These findings are important for the correct interpretation of this widely used medical technique, which may have novel applications in diagnosis and treatment monitoring of aging and skin diseases

Spontaneous BOLD waves – A novel hemodynamic activity in Sprague-Dawley rat brain detected by functional magnetic resonance imaging

We report spontaneous hemodynamic activity termed “Spontaneous BOLD Waves” (SBWs) detected by BOLD fMRI in Sprague-Dawley rats under medetomidine anesthesia. These SBWs, which lasted several minutes, were observed in cortex, thalamus and hippocampus. The SBWs’ correlates were undetectable in electrophysiological recordings, suggesting an exclusive gliovascular phenomenon dissociated from neuronal activity. SBWs were insensitive to the NMDA receptors antagonist MK-801 but were inhibited by the α1-adrenoceptor blocker prazosin. Since medetomidine is a potent agonist of α2 adrenoceptors, we suggested that imbalance in α1/α2 receptor-mediated signalling pathways alter the vascular reactivity leading to SBWs. The frequency of SBWs increased with intensity of mechanical lung ventilation despite the stable pH levels. In summary, we present a novel type of propagating vascular brain activity without easily detectable underlying neuronal activity, which can be utilized to study the mechanisms of vascular reactivity in functional and pharmacological MRI and has practical implications for designing fMRI experiments in anesthetized animals. </jats:p

Time traces of raw PPG signal within the palm area.

<p>(A) An example of the signal during the complete occlusion test (about 12 min). (B) Zoomed part of the signal (blue line) with the temporal boundaries of each cardiac cycle (shown by red circles), which has been approximated to the harmonic reference signal (red dotted line). The black line in the part (B) is the raw PPG signal filtered by continuous averaging of the data over 30 frames.</p

Estimation of the signal-to-noise ratio.

<p>(A) An example of 2D map of the blood pulsation amplitude for subject's arm and the cardboard; (B) Time-traces of the BPA averaged within 15×15 pixels regions shown by black rectangles in (A): the blue line for the arm and the green line for the cardboard; (C – F) Time-averaged BPA from randomly chosen regions within white rectangles sizing 30×30 (C), 15×15 (D), 8×8 (E) pixels, and for single pixel (F).</p