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      Mood-related behavioral and neurochemical alterations in mice exposed to low chlorpyrifos levels during the brain growth spurt

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          Abstract

          Organophosphates are among the most used pesticides. Particularly, chlorpyrifos (CPF) is responsible for a number of deleterious effects on brain development, which may program behavioral changes later in life. Here, we investigated whether a regimen of early low level CPF exposure that did not result in a significant inhibition of acetylcholinesterase (AChE) had deleterious effects on mood-related behaviors, as well as on cholinergic and serotonergic biomarkers in the mice brain. From the 3 rd to 9 th postnatal day (PN), male and female Swiss mice were subcutaneously injected with CPF. Mice were submitted to a battery of behavioral tests from PN60 to PN63: open field, elevated plus maze and forced swimming tests. The cholinergic and serotonergic biomarkers were assessed at PN10 and PN63. Our data indicated that early CPF exposure increased anxiety-like behavior in females and altered decision-making behavior in both sexes. Most biochemical alterations were sex-dependent and restricted to females. At PN10, CPF female mice showed increased serotonin and choline transporter binding in cerebral cortex. Distinctively, in adult females, the effects indicated a hypoactive state: CPF exposure reduced 5-HT 1a receptor binding in cerebral cortex, as well as serotonin transporter binding and choline acetyltransferase activity in brainstem. Our results indicate that CPF exposure during the brain growth spurt deregulates serotonergic and cholinergic biomarkers. The effects are consistent with impaired synaptic function, may be related to long-term mood disorders and point out to higher female susceptibility.

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          Changing numbers of neuronal and non-neuronal cells underlie postnatal brain growth in the rat.

          The rat brain increases >6x in mass from birth to adulthood, presumably through the addition of glial cells and increasing neuronal size, without the addition of neurons. To test this hypothesis, here we investigate quantitatively the postnatal changes in the total number of neuronal and non-neuronal cells in the developing rat brain, and examine how these changes correlate with brain growth. Total numbers of cells were determined with the isotropic fractionator in the brains of 53 Wistar rats, from birth to young adulthood. We find that at birth, >90% of the cells in the rat brain are neurons. Following a dormant period of approximately 3 days after birth, the net number of neurons in the cerebral cortex, hippocampus, and remaining tissue (excluding cerebellum and olfactory bulb) doubles during the first week, then is reduced by 70% during the second postnatal week, concurrently with net gliogenesis. A second round of net addition of 6 million neurons is observed in the cerebral cortex over the following 2 weeks. During the first postnatal week, brain growth relates mainly to increased numbers of neurons of larger average size. In the second and third weeks, it correlates with increased numbers of non-neuronal cells that are smaller in size than the preexisting neurons. Postnatal rat brain development is thus characterized by dramatic changes in the cellular composition of the brain, whose growth is governed by different combinations of cell addition and loss, and changes in average cell size during the first months after birth.
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            Anxiety, defence and the elevated plus-maze.

            The elevated plus-maze test has been in use as a rodent model of anxiety for a decade, and is representative of those tests that are based upon the study of spontaneous behaviour patterns and which have high ecological validity. The origins of the test in studies of the relationship between exploration and fear are reviewed, and attention is drawn to the distinct possibility that variation in the pharmacosensitivity of the procedure may be attributable to often extreme methodological variation between laboratories. In considering further this issue, attention is also drawn to the need to collect data under constant test conditions and to provide the minimum database necessary to reach conclusions regarding the behavioural specificity of drug action. Recent research, which has extended the conventional plus-maze scoring technique to include specific behavioural acts and postures (in particular, those relating to defensive behaviour), is described. The value of such an ethological approach to the plus-maze is then exemplified with original data that demonstrate behaviourally selective, anti-anxiety effects of the GABAA receptor agonist, muscimol (0.125-1.0 mg/kg). It is concluded that, when used appropriately, the elevated plus-maze test can be a very valuable tool in drug screening and in the study of the neurobiology of anxiety and defence. More attention to behaviour and somewhat less emphasis on test simplicity and convenience would seem to be warranted.
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              Elevated anxiety and antidepressant-like responses in serotonin 5-HT1A receptor mutant mice

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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: InvestigationRole: MethodologyRole: Project administrationRole: ValidationRole: VisualizationRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: InvestigationRole: MethodologyRole: Project administrationRole: ValidationRole: VisualizationRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: InvestigationRole: MethodologyRole: Project administrationRole: ValidationRole: VisualizationRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: InvestigationRole: MethodologyRole: Project administrationRole: ValidationRole: VisualizationRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: VisualizationRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: VisualizationRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: VisualizationRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                2 October 2020
                2020
                : 15
                : 10
                : e0239017
                Affiliations
                [1 ] Departamento de Ciências, Faculdade de Formação de Professores da Universidade do Estado do Rio de Janeiro, São Gonçalo, RJ, Brazil
                [2 ] Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
                [3 ] Laboratório de Neurofisiologia, Departamento de Ciências Fisiológicas, Instituto de Biologia Roberto Alcantara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
                [4 ] Instituto de Estudos em Saúde Coletiva e Faculdade de Medicina, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
                Weizmann Institute of Science, ISRAEL
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0003-4324-1413
                Article
                PONE-D-20-18748
                10.1371/journal.pone.0239017
                7531821
                33007016
                13052165-9cc0-4e57-8a2f-1eab5bbf21be
                © 2020 Ribeiro-Carvalho et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 18 June 2020
                : 27 August 2020
                Page count
                Figures: 5, Tables: 2, Pages: 18
                Funding
                This work was supported by grants from Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro - FAPERJ (E26/111.160/2011; E26/103.029/2008 to YA-V, CCF, and ACM), and fellowships from Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq and FAPERJ (CSL); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES (FN, ALN-F). YA-V and AM were Irving J. Selikoff International Scholars of the Mount Sinai School of Medicine. Their work was supported in part by an Award Number D43TW000640 from the Fogarty International Center. The content is solely the responsibility of the authors and does not necessarily represent the official views of the Fogarty International Center or the National Institutes of Health. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
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                Biology and Life Sciences
                Biochemistry
                Neurochemistry
                Neurotransmitters
                Cholinergics
                Biology and Life Sciences
                Neuroscience
                Neurochemistry
                Neurotransmitters
                Cholinergics
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                Biochemistry
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                Neurotransmitters
                Biogenic Amines
                Serotonin
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                All data files are available in the Mendeley database. http://dx.doi.org/10.17632/9bf53dvvp7.1.

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