Selenoneine is a major selenium species in beluga skin and red blood cells of Inuit from Nunavik

Nunavimmiut (Inuit of Nunavik, Northern Quebec, Canada) exhibit a high selenium (Se) status because oftheir frequent consumption of marine mammal foods. Indirect evidence from our previous studies hadsuggested that selenoneineea novel selenocompoundemay be accumulating in the blood of Nuna-vimmiut. We used a liquid-chromatography/inductively coupled tandem mass spectrometry (LC-ICP-MS/MS) method to measure concentrations of selenoneine and its methylated metabolite Se-methylselenoneine in archived red blood cells (RBC) obtained from 210 Nunavimmiut living in com-munities along the Hudson Strait, where marine mammal hunting and consumption are most frequent inNunavik. This method was adapted to quantify selenoneine and its methylated metabolite in belugamattaaq, an Inuit delicacy consisting of the skin with the underlying layer of fat and the major dietarysource of Se for Nunavimmiut. Total selenium concentration was also measured in RBC and belugamattaaq samples by isotope dilution ICP-MS/MS. The median selenoneine concentration in RBC was413mg Se/L (range¼3.20e3230mg Se/L), representing 54% (median) of total Se content (range¼1.6e91%). Quantification of selenoneine infive beluga mattaaq samples (skin layer) from Nunavik revealed amedian concentration of 1.8mg Se/g wet wt (range¼1.2e7.4mg Se/g), constituting 54% (median) of thetotal Se content (range¼44e74%). Se-methylselenoneine was also detected in Inuit RBC but not inbeluga mattaaq, suggesting that selenoneine undergoes methylation in humans. Selenoneine may protect Nunavimmiut from methylmecury toxicity by increasing its demethylation in RBC and in turndecreasing its distribution to target organs

Stimulation optique localisée assistée par les nanoparticules d’or : un nouvel outil pour étudier la communication synaptique et la plasticité

Pour mieux comprendre les processus moléculaires à la base de la communication synaptique, il est nécessaire de pouvoir observer et contrôler l’activité synaptique avec une grande précision temporelle (ms) et spatiale (échelle nanométrique). Mon projet propose d’utiliser une technique de stimulation optique localisée basée sur l’excitation plasmonique de nanoparticules d’or (Nanoparticle Assisted Localized Optical Stimulation – NALOS) afin de produire une stimulation synaptique localisée sur une seule synapse. NALOS peut être réalisé en utilisant un microscope confocal équipé d’un laser infrarouge femtoseconde pour stimuler des nanoparticules d’or déposées sur les neurones en culture. Il a été démontré qu’il est possible d’induire avec NALOS une augmentation transitoire de Ca2+ intracellulaire sur une dendrite, mesurée avec l’aide de GCaMP6s, un indicateur de Ca2+ génétiquement encodé1 . Afin de mieux comprendre les effets physiologiques de NALOS sur les neurones, la première partie de mon projet vise à caractériser le mécanisme sous-jacent à la technique. Pour ce faire, nous avons varié la puissance de la stimulation laser sur les nanoparticules d’or pour caractériser les différentes réponses calciques transitoires intracellulaires obtenues ainsi que la possible formation de trous sur la membrane. Nous avons ensuite identifié les principaux récepteurs et canaux ioniques pouvant être stimulés avec NALOS en utilisant différents antagonistes. Nous avons ensuite appliqué NALOS pour générer une stimulation synaptique. Pour y parvenir, nous avons stimulé localement un axone et observé la réponse calcique reliée à une stimulation synaptique par relâchement naturel de glutamate via des vésicules de la zone active présynaptique. Cet outil permettra donc de stimuler et d’observer, une synapse à la fois, des changements structurels et moléculaires reliés à la communication synaptique.To better understand the molecular processes underlying synaptic communication, it is necessary to be able to observe and control synaptic activity with high temporal (ms) and spatial (nanoscale) precision. My project proposes using a localized optical stimulation technique based on the plasmonic excitation of gold nanoparticles (Nanoparticle Assisted Localized Optical Stimulation – NALOS) to produce synaptic stimulation localized at a single synapse. NALOS can be performed using a confocal microscope equipped with an infrared femtosecond laser to stimulate gold nanoparticles deposited on cultured neurons. It has been shown that NALOS can induce a transient stimulation of intracellular Ca2+ on a dendrite, measured with the help of GCaMP6s, a genetically encoded Ca2+ indicator1 . For a better understanding the physiological effects of NALOS on neurons, the first part of my project aims to characterize the mechanism underlying this technique. To do this, we varied the power of laser stimulation on gold nanoparticles to decipher the different intracellular transient Ca2+ responses obtained as well as to investigate the possible formation of holes on the membrane. We then determined the main receptors and ion channels that can be stimulated with NALOS. We then applied NALOS to generate synaptic stimulation. To do this, we locally stimulated an axon and look at the Ca2+ response related to synaptic stimulation by natural release of glutamate via vesicles of the presynaptic active zone. This tool will thus make it possible to stimulate and observe, one synapse at a time, structural and molecular changes related to synaptic communicatio

Determinants of selenoneine concentration in red blood cells of Inuit from Nunavik (Northern Québec, Canada)

Selenium (Se) is a trace mineral essential to human health, and is especially abundant in marine foods consumed by Inuit populations in Nunavik (northern Quebec, Canada), leading to exceptionally high whole blood Se levels. While most epidemiological studies to date examine plasma or whole blood Se, little is known about the health implications of specific Se biomarkers (e.g. selenoproteins and small Se compounds). Selenoneine, a novel Se compound, is found in high concentrations in marine foods (and particularly beluga mattaaq) and the red blood cells (RBCs) of populations that consume them. We report here RBC selenoneine concentrations in a population of Inuit adults (n = 885) who participated in the Qanuippitaa? 2004 survey. Simple associations between RBC selenoneine and other Se and mercury (Hg) biomarkers were assessed using Spearman correlations and linear regressions. Wilcoxon ranksum tests were used to examine differences in biomarkers and characteristics between tertiles of RBC selenoneine concentration. A multiple linear regression analysis was used to determine factors (sociodemographic, lifestyle, and dietary) associated with RBC selenoneine concentrations. Selenoneine comprised a large proportion of whole blood Se and RBC Se in this population. Age and sex-adjusted geometric mean RBC selenoneine concentration was 118 μg/L (range: 1-3226 μg/L) and was much higher (p = 0.001) among women (150.3 μg/L) than men (87.6 μg/L) across all regions of Nunavik after controlling for age, region, and diet. RBC selenoneine was highly correlated with RBC Se (rs = 0.96, p < 0.001) and whole blood Se (rs = 0.89, p < 0.001), but only weakly correlated with plasma Se (rs = 0.13, p < 0.001). Overall, increasing age (standardized β = 0.24), higher body-mass index (BMI; β = 0.08), female sex (β = 0.10), living in a Hudson Strait community (compared to Hudson Bay and Ungava Bay; β = 0.38), and consuming beluga mattaaq (g/day; β = 0.19) were positively associated with RBC selenoneine. Meanwhile, consumption of market meats (g/day; β = -0.07) was negatively associated with RBC selenoneine. RBC selenoneine is an important biomarker of Se dietary intake from local marine foods in Inuit populations. Further studies are needed to examine the health effects of selenoneine intake and the underlying mechanisms for sex differences among Inuit populations