RESUMO
Alcohol affects multiple neurotransmitter systems, notably the GABAergic system and has been recognised for a long time as particularly damaging during critical stages of brain development. Nevertheless, data from the literature are most often derived from animal or in vitro models. In order to study the production, migration and cortical density disturbances of GABAergic interneurons upon prenatal alcohol exposure, we performed immunohistochemical studies by means of the proliferation marker Ki67, GABA and calretinin antibodies in the frontal cortical plate of 17 foetal and infant brains antenatally exposed to alcohol, aged 15 weeks' gestation to 22 postnatal months and in the ganglionic eminences and the subventricular zone of the dorsal telencephalon until their regression, i.e., 34 weeks' gestation. Results were compared with those obtained in 17 control brains aged 14 weeks of gestation to 35 postnatal months. We also focused on interneuron vascular migration along the cortical microvessels by confocal microscopy with double immunolabellings using Glut1, GABA and calretinin. Semi-quantitative and quantitative analyses of GABAergic and calretininergic interneuron density allowed us to identify an insufficient and delayed production of GABAergic interneurons in the ganglionic eminences during the two first trimesters of the pregnancy and a delayed incorporation into the laminar structures of the frontal cortex. Moreover, a mispositioning of GABAergic and calretininergic interneurons persisted throughout the foetal life, these cells being located in the deep layers instead of the superficial layers II and III. Moreover, vascular migration of calretininergic interneurons within the cortical plate was impaired, as reflected by low numbers of interneurons observed close to the cortical perforating vessel walls that may in part explain their abnormal intracortical distribution. Our results are globally concordant with those previously obtained in mouse models, in which alcohol has been shown to induce an interneuronopathy by affecting interneuron density and positioning within the cortical plate, and which could account for the neurological disabilities observed in children with foetal alcohol disorder spectrum.
Assuntos
Consumo de Bebidas Alcoólicas , Encéfalo/embriologia , Calbindina 2/metabolismo , Transtornos do Espectro Alcoólico Fetal/metabolismo , Feto/embriologia , Interneurônios/metabolismo , Antígeno Ki-67/metabolismo , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Ácido gama-Aminobutírico/metabolismo , Alcoolismo , Consumo Excessivo de Bebidas Alcoólicas , Encéfalo/metabolismo , Encéfalo/patologia , Estudos de Casos e Controles , Movimento Celular , Feminino , Transtornos do Espectro Alcoólico Fetal/patologia , Feto/metabolismo , Feto/patologia , Lobo Frontal/embriologia , Lobo Frontal/metabolismo , Lobo Frontal/patologia , Neurônios GABAérgicos/metabolismo , Neurônios GABAérgicos/patologia , Humanos , Lactente , Recém-Nascido , Interneurônios/patologia , Masculino , Gravidez , Complicações na Gravidez , Segundo Trimestre da Gravidez , Efeitos Tardios da Exposição Pré-Natal/patologia , Telencéfalo/embriologia , Telencéfalo/metabolismo , Telencéfalo/patologiaRESUMO
The contamination of foods with mineral oil hydrocarbons (MOH) is a serious concern, requiring in most cases tedious mitigation measures that span across the whole food supply chain. A major issue today is the significant variability of the results generated by laboratories. This study was therefore designed to achieve a deeper insight into the analytical procedures used by commercial laboratories, identifying possible gaps and suggesting improvements that will enhance the reliability of the MOH data, an important prerequisite for risk assessment. In total six different food matrices, i.e. infant formula (IF), cocoa butter, cocoa powder, biscuits, fruit-based baby food containing biscuit and roast and ground coffee were subjected to comparative inter-laboratory studies, as well as one vegetable oil analysed within the frame of a professionally conducted proficiency test. The results indicate that on some matrices with possibly low amounts of MOH contamination, the current methodologies cannot reliably conclude whether or not a food sample is indeed contaminated with mineral oils (<10 mg/kg food). Urgently needed are: (i) an aligned and fully validated sample preparation strategy tested on a range of different food matrices; (ii) a confirmation of positive flame ionisation detection (FID) results by confirmatory methods such as mass spectrometry - in line with the CEN Standard and the Joint Research Centre (JRC) Guidance Document, (iii) a more detailed root-cause analysis in the reports of laboratories through the use of mineral oil markers, and (iv) a fully validated official method for the concerned foods with a limit of application <10 mg/kg food.