When it comes to cell types, the cerebral cortex is one of the most diverse regions in the mammalian brain. Mouse cortical neurons are generated during development from radial glial cells (RGCs). But how these stem cells generate the different neuronal subtypes is still an open question. In the adult, transcription factors, specially homeobox genes, have been identified as determinants of neuronal types throughout the animal kingdom. Thus, in this study, we hypothesise that different subpopulations of neuronal progenitors (RGCs) give rise to subsequent subtypes of neurons in the cortex, and these populations can be defined by homeobox gene expression. Starting from a scRNA- seq analysis, we identified differentially expressed genes across different progenitor populations in the developing cortex: Adnp2, Homez and Hmbox1. We characterised their mRNA and protein expression across cortical layers in postnatal mice and found that these genes are also differentially expressed among layers. We also find discordances between scRNA-seq data, mRNA expression, and protein expression data that could indicate specific post-transcriptional regulation of these genes. Altogether, these results point to a role of homeobox genes in neuronal identity.

No / Cortical spreading depression (CSD) is a temporary disruption of local ionic homeostasis that propagates slowly across the cerebral cortex, and may contribute to the pathophysiology of stroke and migraine. Previous studies demonstrated that nitric oxide (NO) formation promotes the repolarisation phase of CSD, and this effect may be cyclic GMP (cGMP)-mediated. Here, we have examined how phosphodiesterase (PDE) inhibition, either alone or superimposed on NO synthase (NOS) inhibition, alters CSD and the associated changes in extracellular cGMP. Microdialysis probes incorporating an electrode were implanted into the frontoparietal cortex of anaesthetised rats for quantitative recording of CSD, pharmacological manipulations, and dialysate sampling for cGMP measurements. CSD was induced by cathodal electrical stimulation in the region under study by microdialysis. Extracellular cGMP increased, but only slightly, during CSD. Perfusion of either zaprinast or sildenafil through the microdialysis probe, at concentrations that inhibited both PDE5 and PDE9 (and possibly other PDE), increased significantly extracellular cGMP. Unexpectedly, these levels remained high when NOS was subsequently inhibited with N¿-nitro- -arginine methyl ester hydrochloride ( -NAME, 1 mM). The most interesting pharmacological effect on CSD was obtained with sildenafil. This drug altered neither CSD nor the subsequent characteristic effect of NOS inhibition, i.e. a marked widening of CSD. The fact that NOS inhibition still widened CSD in the presence of the high extracellular levels of cGMP associated with PDE inhibition, suggests that NO may promote CSD recovery, independently of cGMP formation.

No / Preconditioning of the cerebral cortex was induced in mice by repeated cortical spreading depression (CSD), and the major ionotropic glutamate (GluRs) and nicotinic acetylcholine receptor (nAChRs) subunits were compared by quantitative immunoblotting between sham- and preconditioned cortex, 24 h after treatment. A 30% reduction in ¿-amino-3-hydroxy-5-methyl-4-iso- xazolepropionate (AMPA) GluR1 and 2 subunit immunoreactivities was observed in the preconditioned cortex (p < 0.03), but there was no significant change in the NMDA receptor subunits, NR1, NR2A and NR2B. A 12¿15-fold increase in ¿7 nAChR subunit expression following in vivo CSD (p < 0.001) was by far the most remarkable change associated with preconditioning. In contrast, the ¿4 nAChR subunit was not altered. These data point to the ¿7 nAChR as a potential new target for neuroprotection because preconditioning increases consistently the tolerance of the brain to acute insults such as ischaemia. These data complement recent studies implicating ¿7 nAChR overexpression in the amelioration of chronic neuropathologies, notably Alzheimer's disease (AD).

No / Human migration is a hallmark of the species and there is significant interest in methods that can determine the past migrations of humans and associated fauna. We present a new method that utilizes collagen oxygen, carbon and nitrogen isotopes from histologically informed samples of cortical bone. The utility of this multi-isotopic, life history approach is demonstrated in migrating Dutch whalers, and both the possibilities and limitations of the method are described.

Yes / Despite intense attention from biomedical and chemical researchers, there are few approved treatments for amyotrophic lateral sclerosis (ALS), with only riluzole (Rilutek) and edaravone (Radicava) currently available to patients. Moreover, the mechanistic basis of the activity of these drugs is currently not well-defined, limiting the ability to design new medicines for ALS. This Letter describes the synthesis of triazole-containing riluzole analogues, and their testing in a novel neuroprotective assay. Seven compounds were identified as having neuroprotective activity, with two compounds having similar activity to riluzole.

Yes / Many visual effects are believed to be processed at several functional and anatomical levels of cortical processing. Determining if and how the levels contribute differentially to these effects is a leading problem in visual perception and visual neuroscience. We review and analyze a combination of extant psychophysical findings in the context of neurophysiological and brain-imaging results. Specifically using findings relating to visual illusions, crowding, and masking as exemplary cases, we develop a theoretical rationale for showing how relative levels of cortical processing contributing to these effects can already be deduced from the psychophysically determined functions relating respectively the illusory, crowding and masking strengths to the contrast of the illusion inducers, of the flankers producing the crowding, and of the mask. The wider implications of this rationale show how it can help to settle or clarify theoretical and interpretive inconsistencies and how it can further psychophysical, brain-recording and brain-imaging research geared to explore the relative functional and cortical levels at which conscious and unconscious processing of visual information occur. Our approach also allows us to make some specific predictions for future studies, whose results will provide empirical tests of its validity.

No description available.

La division cellulaire asymétrique (DCA) consiste en une division pendant laquelle des déterminants cellulaires sont distribués préférentiellement dans une des deux cellules filles. Par l’action de ces déterminants, la DCA générera donc deux cellules filles différentes. Ainsi, la DCA est importante pour générer la diversité cellulaire et pour maintenir l’homéostasie de certaines cellules souches. Pour induire une répartition asymétrique des déterminants cellulaires, le positionnement du fuseau mitotique doit être très bien contrôlé. Fréquemment ceci génère deux cellules filles de tailles différentes, car le fuseau mitotique n’est pas centré pendant la mitose, ce qui induit un positionnement asymétrique du sillon de clivage. Bien qu’un complexe impliquant des GTPases hétérotrimériques et des protéines liant les microtubules au cortex ait été impliqué directement dans le positionnement du fuseau mitotique, le mécanisme exact induisant le positionnement asymétrique du fuseau durant la DCA n'est pas encore compris. Des études récentes suggèrent qu’une régulation asymétrique du cytosquelette d’actine pourrait être responsable de ce positionnement asymétrique du faisceau mitotique. Donc, nous émettons l'hypothèse que des contractions asymétriques d’actine pendant la division cellulaire pourraient déplacer le fuseau mitotique et le sillon de clivage pour créer une asymétrie cellulaire. Nos résultats préliminaires ont démontré que le blebbing cortical, qui est une indication de tension corticale et de contraction, se produit préférentiellement dans la moitié antérieure de cellule précurseur d’organes sensoriels (SOP) pendant le stage de télophase. Nos données soutiennent l'idée que les petites GTPases de la famille Rho pourraient être impliqués dans la régulation du fuseau mitotique et ainsi contrôler la DCA des SOP. Les paramètres expérimentaux développés pour cette thèse, pour étudier la régulation de l’orientation et le positionnement du fuseau mitotique, ouvrirons de nouvelles avenues pour contrôler ce processus, ce qui pourrait être utile pour freiner la progression de cellules cancéreuses. Les résultats préliminaires de ce projet proposeront une manière dont les petites GTPases de la famille Rho peuvent être impliqués dans le contrôle de la division cellulaire asymétrique in vivo dans les SOP. Les modèles théoriques qui sont expliqués dans cette étude pourront servir à améliorer les méthodes quantitatives de biologie cellulaire de la DCA. / Asymmetric cell division (ACD) consists in a cellular division during which specific cell fate determinants are distributed preferentially in one daughter cell, which then differentiate from its sibling. Hence, ACD is important to generate cell diversity and is used to regulate stem cells homeostasis. For proper asymmetric distribution of cell fate determinants, the positioning of the mitotic spindle has to be tightly controlled. Frequently, this induces a cell size asymmetry, since the spindle is then not centered during mitosis, leading to an asymmetric positioning of the cleavage furrow. Although small small GTPases have been shown to act directly on the spindle, the exact mechanism controlling spindle positioning during ACD is not understood. Recent studies suggest that an independent, yet uncharacterized pathway is involved in spindle positioning, which is likely to involve an asymmetric regulation of the actin cytoskeleton. Indeed, actin enables spindle anchoring to the cortex. Hence we hypothesize that asymmetric actin contractions during cytokinesis might displace the mitotic spindle and the cleavage furrow, leading to cell size asymmetry. Interestingly, from our preliminary results we observed that cortical blebbing, which is a read-out of cortical tension/contraction, preferentially occurs on the anterior side of the dividing sensory organ precursor (SOP) cells at telophase. Our preliminary data support the idea that Rho small GTPases might be implicated in regulation of the mitotic spindle hence controlling asymmetric cell division of SOP cells. The experimental settings developed for this thesis, for studying regulation of the mitotic spindle orientation and positioning will serve as proof of concept of how geneticist and biochemist experts could design ways to control such process by different means in cancerous cells. The preliminary results from this project open novel insights on how the Rho small GTPases might be implicated in controlling asymmetric cell division hence their dynamics in vivo of such process during SOP development. Furthermore, the assays and the theoretical model developed in this study can be used as background that could serve to design improved quantitative experimental methods for cell biology synchronizing sub-networks of ACD mechanism.

The subplate layer of the cerebral cortex is comprised of a heterogeneous population of cells and contains some of the earliest-generated neurons. Subplate plays a fundamental role in cortical development. In the embryonic brain, subplate cells contribute to the guidance and areal targeting of corticofugal and thalamic axons. At later stages, these cells are involved in the maturation and plasticity of the cortical circuitry and the establishment of functional modules. In my thesis, I aimed to further characterize the embryonic murine subplate by establishing a gene expression profile of this population at embryonic day 15.5 (E15.5) using laser capture microdissection combined with microarrays. I found over 250 transcripts with presumed higher expression in the subplate at E15.5. Using quantitative RT-PCR, in situ hybridization and immunohistochemistry, I have confirmed specific expression in the E15.5 subplate for 13 selected genes which have not been previously associated with this compartment. In the reeler mutant, the expression pattern of a majority of these genes was shifted in accordance with the altered position of subplate cells. These genes belong to several functional groups and likely contribute to the maturation and electrophysiological properties of subplate cells and to axonal growth and guidance. The roles of two selected genes - cadherin 10 (Cdh10) and Unc5 homologue c (Unc5c) - were explored in more detail. Preliminary results suggest an involvement of Cdh10 in subplate layer organization while Unc5c could mediate the waiting period of subplate corticothalamic axons in the internal capsule. Finally, I compared the expression of a selection of subplate-specific genes (subplate markers) between mouse and rat and found some surprising species differences. Confirmed subplate markers were used to monitor subplate injury in a rat model of preterm hypoxiaischemia and it appeared that deep cortical layers including subplate showed an increased vulnerability over upper layers. Further characterization of subplate-specific genes will allow us to broaden our understanding of molecular mechanisms underlying subplate properties and functions in normal and pathological development.

Orientador: Guilherme de Paula Nogueira / Banca: Yuri Tani Utsunomiya / Banca: Marco Antonio Rodrigues Fernandes / Resumo: O objetivo desse estudo foi inferir a Densidade Mineral óssea (DMO) a partir da imagem radiográfica (Raios-X) usando como referência a DMO de uma escada de alumínio mensurada por Absorciometria por de Duas-Energias (DXA). Para isso foram utilizadas 30 amostras de tecido ósseo cortical e 30 amostras de tecido ósseo trabecular de osso bovino "in vitro". Foram mensurados neste estudo a Densidade Mineral Óssea (DMO), o Conteúdo Mineral Ósseo (CMO) ambos obtidos pelo densitômetro LUNAR®-DPX ALPHA; a Quantidade Mineral Óssea (QMO) massa das cinzas após a calcinação das amostras; a densidade real das amostras (dReal=massa/volume). Foram feitas 10 tomadas de RX com as 60 amostras, entremeadas pela escada de alumínio (referencial densitométrico). A correlação entre as técnicas (DR-tons de cinza e DXA g/cm2) gerou equações de regressão para cada uma das dez radiografias e permitiu inferir a densidade mineral óssea (DMODR), obtida através da conversão dos tons de cinza em densidade radiográfica pelo software ImageJ® para cada uma das 10 tomadas radiográficas; calculou-se então a média da densidade mineral óssea calculada pela densitometria radiográfica (XDMODR) de todas as dez radiografias. Foi observado que a média da densidade real das amostras foi de 2,2±0,23g/cm2 e 1,05±0,09g/cm2 enquanto que a média da densidade mensurada pelo DXA foi de 0,73±0,11g/cm2 e 0,22±0,11g/cm2, observou que a media da DMODR ficou em 0,93±0,11g/cm2 e 0,34±0,14 g/cm2 para o osso cortical e trabecular respect... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The aim of this study was to infer the bone mineral density (BMD) from the radiographic image (X-X) with reference to the BMD of an aluminum step wedge measured by absorptiometry Dual-energy (DXA). For this we used 30 samples of cortical bone 30 and cancellous bone tissue samples of bovine bone "in vitro". Were measured in this study Bone Mineral Density (BMD), bone mineral content (BMC) both obtained by densitometer LUNAR® DPX-ALPHA; Volume Bone Mineral (QMO) mass of ash after calcination of the samples; the real density of the samples (dReal = mass / volume). 10 taken RX were made with the samples 60, interspersed by aluminum step wedge (densitometric reference). The correlation between the techniques (gray DR-tones and DXA g / cm 2) generated regression equations for each of the ten X-rays and allowed to infer bone mineral density (DMODR) obtained by converting grayscale radiographic density by ImageJ® for each of the 10 radiographic taken. It was then calculated average bone mineral density calculated by X-ray densitometry (XDMODR) radiographs of all ten. Was observed that the average true density of the samples was 2.2±0.23g/cm2 and 1.05±0.09g/cm2 while the average density measured by DXA was 0.73±0.11g/cm2 and 0.22±0.11g/cm2 interesting that the average DMODR was 0.93±0.11g/cm2 and 0.34±0.14g/cm2 for cortical and cancellous bone respectively. The average of the BMC was 0.19±0.08g and 0.9±0.02g, and the average QMO 0.14±0.04g and 0.05±0 03g for cortical and cancellous bo... (Complete abstract click electronic access below) / Mestre