Elevated P75NTR expression causes death of engrailed-deficient midbrain dopaminergic neurons by Erk1/2 suppression

<p>Abstract</p> <p>Background</p> <p>The homeodomain transcription factors <it>Engrailed-1 </it>and <it>Engrailed-2 </it>are required for the survival of mesencephalic dopaminergic (mesDA) neurons in a cell-autonomous and gene-dose-dependent manner. Homozygote mutant mice, deficient of both genes (<it>En1-/-;En2-/-</it>), die at birth and exhibit a loss of all mesDA neurons by mid-gestation. In heterozygote animals (<it>En1+/-;En2-/-</it>), which are viable and fertile, postnatal maintenance of the nigrostriatal dopaminergic system is afflicted, leading to a progressive degeneration specific to this subpopulation and Parkinson's disease-like molecular and behavioral deficits.</p> <p>Results</p> <p>In this work, we show that the dose of <it>Engrailed </it>is inversely correlated to the expression level of the pan-neurotrophin receptor gene <it>P75</it>^{<it>NTR </it>}(<it>Ngfr</it>). Loss of mesDA neurons in the <it>Engrailed</it>-null mutant embryos is caused by elevated expression of this neurotrophin receptor: Unusually, in this case, the cell death signal of P75^NTRis mediated by suppression of Erk1/2 (extracellular-signal-regulated kinase 1/2) activity. The reduction in expression of <it>Engrailed</it>, possibly related to the higher levels of P75^NTR, also decreases mitochondrial stability. In particular, the dose of <it>Engrailed </it>determines the sensitivity to cell death induced by the classic Parkinson-model toxin MPTP and to inhibition of the anti-apoptotic members of the Bcl-2 family of proteins.</p> <p>Conclusion</p> <p>Our study links the survival function of the <it>Engrailed </it>genes in developing mesDA neurons to the regulation of <it>P75</it>^{<it>NTR </it>}and the sensitivity of these neurons to mitochondrial insult. The similarities to the disease etiology in combination with the nigral phenotype of <it>En1+/-;En2-/- </it>mice suggests that haplotype variations in the <it>Engrailed </it>genes and/or <it>P75</it>^{<it>NTR </it>}that alter their expression levels could, in part, determine susceptibility to Parkinson's disease.</p

Embryonic Exposure to Valproic Acid Affects Social Predispositions for Dynamic Cues of Animate Motion in Newly-Hatched Chicks

Early predispositions to preferentially orient towards cues associated with social partners have been documented in several vertebrate species including human neonates and domestic chicks. Human newborns at high familiar risk of Autism Spectrum Disorder (ASD) show differences in their attention toward these predisposed stimuli, suggesting potential impairments in these social-orienting mechanisms in ASD. Using embryonic exposure to valproic acid (VPA) we modelled ASD behavioural deficits in domestic chicks. To investigate social predispositions towards animate motion in domestic chicks, we focused on self-propulsion, using two video-animations representing a simple red circle moving at constant speed (speed-constant) or one that was changing its speed (accelerating and decelerating; speed-change). Using a six minutes spontaneous choice test for the two stimuli, we compared unlearned preferences for stimuli that autonomously change speed between VPA- and vehicle-injected chicks. We found that the preference for speed changes was abolished in VPA-injected chicks compared to vehicle-injected controls. These results add to previous findings indicating similar impairments for static social stimuli and suggest a specific effect of VPA on the development of mechanisms that enhance orienting towards animate stimuli. These findings strengthen the hypothesis of an early impairment of predispositions in the early development of ASD. Hence, early predispositions are a potentially useful tool to detect early ASD symptoms in human neonates and to investigate the molecular and neurobiological mechanisms underlying the onset of this neurodevelopmental disorder

Fetal blockade of nicotinic acetylcholine transmission causes autism-like impairment of biological motion preference in the neonatal chick

Several environmental chemicals are suspected risk factors for autism spectrum disorder (ASD), including valproic acid (VPA) and pesticides acting on nicotinic acetylcholine receptors (nAChRs), if administered during pregnancy. However, their target processes in fetal neuro-development are unknown. We report that the injection of VPA into the fetus impaired imprinting to an artificial object in neonatal chicks, while a predisposed preference for biological motion (BM) remained intact. Blockade of nAChRs acted oppositely, sparing imprinting and impairing BM preference. Beside ketamine and tubocurarine, significant effects of imidacloprid (a neonicotinoid insecticide) appeared at a dose ≤1 ppm. In accord with the behavioral dissociations, VPA enhanced histone acetylation in the primary cell culture of fetal telencephalon, whereas ketamine did not. VPA reduced the brain weight and the ratio of NeuN-positive cells (matured neurons) in the telencephalon of hatchlings, whereas ketamine/tubocurarine did not. Despite the distinct underlying mechanisms, both VPA and nAChR blockade similarly impaired imprinting to biological image composed of point-light animations. Furthermore, both impairments were abolished by postnatal bumetanide treatment, suggesting a common pathology underlying the social attachment malformation. Neurotransmission via nAChR is thus critical for the early social bond formation, which is hindered by ambient neonicotinoids through impaired visual predispositions for animate objects

Physiological and behavioral responses in Drosophila melanogaster to odorants present at different plant maturation stages

The fruit fly Drosophila melanogaster feeds and oviposits on fermented fruit, hence its physiological and behavioral responses are expected to be tuned to odorants abundant during later stages of fruit maturation. We used a population of about two-hundred isogenic lines of D. melanogaster to assay physiological responses (electroantennograms (EAG)) and behavioral correlates (preferences and choice ratio) to odorants found at different stages of fruit maturation. We quantified electrophysiological and behavioral responses of D. melanogaster for the leaf compound β-cyclocitral, as well as responses to odorants mainly associated with later fruit maturation stages. Electrophysiological and behavioral responses were modulated by the odorant dose. For the leaf compound we observed a steep dose-response curve in both EAG and behavioral data and shallower curves for odorants associated with later stages of maturation. Our data show the connection between sensory and behavioral responses and are consistent with the specialization of D. melanogaster on fermented fruit and avoidance of high doses of compounds associated with earlier stages of maturation. Odor preferences were modulated in a non-additive way when flies were presented with two alternative odorants, and combinations of odorants elicited higher responses than single compounds

Valproic acid exposure affects social visual lateralization and asymmetric gene expression in zebrafish larvae

Abstract Cerebral asymmetry is critical for typical brain function and development; at the same time, altered brain lateralization seems to be associated with neuropsychiatric disorders. Zebrafish are increasingly emerging as model species to study brain lateralization, using asymmetric development of the habenula, a phylogenetically old brain structure associated with social and emotional processing, to investigate the relationship between brain asymmetry and social behavior. We exposed 5-h post-fertilization zebrafish embryos to valproic acid (VPA), a compound used to model the core signs of ASD in many vertebrate species, and assessed social interaction, visual lateralization and gene expression in the thalamus and the telencephalon. VPA-exposed zebrafish exhibit social deficits and a deconstruction of social visual laterality to the mirror. We also observe changes in the asymmetric expression of the epithalamic marker leftover and in the size of the dorsolateral part of the habenula in adult zebrafish. Our data indicate that VPA exposure neutralizes the animals’ visual field bias, with a complete loss of the left-eye use bias in front of their own mirror image, and alters brain asymmetric gene expression and morphology, opening new perspectives to investigate brain lateralization and its link to atypical social cognitive development

Altered Expression of GABAergic Markers in the Forebrain of Young and Adult Engrailed-2 Knockout Mice

Impaired function of GABAergic interneurons, and the subsequent alteration of excitation/inhibition balance, is thought to contribute to autism spectrum disorders (ASD). Altered numbers of GABAergic interneurons and reduced expression of GABA receptors has been detected in the brain of ASD subjects and mouse models of ASD. We previously showed a reduced expression of GABAergic interneuron markers parvalbumin (PV) and somatostatin (SST) in the forebrain of adult mice lacking the Engrailed2 gene (En2-/- mice). Here, we extended this analysis to postnatal day (P) 30 by using in situ hybridization, immunohistochemistry, and quantitative RT-PCR to study the expression of GABAergic interneuron markers in the hippocampus and somatosensory cortex of En2-/- and wild type (WT) mice. In addition, GABA receptor subunit mRNA expression was investigated by quantitative RT-PCR in the same brain regions of P30 and adult En2-/- and WT mice. As observed in adult animals, PV and SST expression was decreased in En2-/- forebrain of P30 mice. The expression of GABA receptor subunits (including the ASD-relevant Gabrb3) was also altered in young and adult En2-/- forebrain. Our results suggest that GABAergic neurotransmission deficits are already evident at P30, confirming that neurodevelopmental defects of GABAergic interneurons occur in the En2 mouse model of ASD

Light-induced asymmetries in embryonic retinal gene expression are mediated by the vascular system and extracellular matrix

Funding: This project was supported by BBSRC Grant BB/S003223/1 (EV, JG), Royal Society Grant UF150663 (SP), Novo Nordisk Foundation Grant NNF14CC0001 (LJJ), ERC Horizon 2020 Grant Agreement 833504 SPANUMBRA, PRIN 2017 and ERC-SH4-A (2017PSRHPZ) to GV.Left–right asymmetries in the nervous system (lateralisation) influence a broad range of behaviours, from social responses to navigation and language. The role and pathways of endogenous and environmental mechanisms in the ontogeny of lateralisation remains to be established. The domestic chick is a model of both endogenous and experience-induced lateralisation driven by light exposure. Following the endogenous rightward rotation of the embryo, the asymmetrical position in the egg results in a greater exposure of the right eye to environmental light. To identify the genetic pathways activated by asymmetric light stimulation, and their time course, we exposed embryos to different light regimes: darkness, 6 h of light and 24 h of light. We used RNA-seq to compare gene expression in the right and left retinas and telencephalon. We detected differential gene expression in right vs left retina after 6 h of light exposure. This difference was absent in the darkness condition and had already disappeared by 24 h of light exposure, suggesting that light-induced activation is a self-terminating phenomenon. This transient effect of light exposure was associated with a downregulation of the sensitive-period mediator gene DIO2 (iodothyronine deiodinase 2) in the right retina. No differences between genes expressed in the right vs. left telencephalon were detected. Gene networks associated with lateralisation were connected to vascularisation, cell motility, and the extracellular matrix. Interestingly, we know that the extracellular matrix—including the differentially expressed PDGFRB gene—is involved in morphogenesis, sensitive periods, and in the endogenous chiral mechanism of primary cilia, that drives lateralisation. Our data show a similarity between endogenous and experience-driven lateralisation, identifying functional gene networks that affect lateralisation in a specific time window.Publisher PDFPeer reviewe