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      Possible role of NO/NMDA pathway in the autistic-like behaviors induced by maternal separation stress in mice

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          Abstract

          Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder. Maternal separation (MS) stress is an established model of early-life stress associated with autistic-like behaviors. Altered glutamatergic and nitrergic neurotransmissions may contribute to the pathophysiology of ASD. However, the specific mechanisms underlying these alterations and their relationship to MS-induced autistic-like behaviors remain unclear. Addressing this knowledge gap, this study aims to elucidate the involvement of the nitric oxide (NO)/ N-methyl-D-aspartate (NMDA) pathway in MS-induced autistic-like behaviors in mice. This knowledge has the potential to guide future research, potentially leading to the development of targeted interventions or treatments aimed at modulating the NO/NMDA pathway to ameliorate ASD symptoms. Ninety male Naval Medical Research Institute (NMRI) mice were assigned to six groups (n = 15) comprising a control group (treated with saline) and five groups subjected to MS and treated with saline, ketamine, NMDA, L-NAME, and L-arginine. Behavioral tests were conducted, including the three-chamber test, shuttle box, elevated plus-maze, and marble burying test. Gene expression of iNOS, nNOS, and NMDA-R subunits (NR2A and NR2B), along with nitrite levels, was evaluated in the hippocampus. The findings demonstrated that MS induced autistic-like behaviors, accompanied by increased gene expression of iNOS, nNOS, NR2B, NR2A, and elevated nitrite levels in the hippocampus. Modulation of the NO/NMDA pathway with activators and inhibitors altered the effects of MS. These results suggest that the NO/NMDA pathway plays a role in mediating the negative effects of MS and potentially contributes to the development of autistic-like behaviors in maternally separated mice.

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          NMDA receptor subunit diversity: impact on receptor properties, synaptic plasticity and disease.

          NMDA receptors (NMDARs) are glutamate-gated ion channels and are crucial for neuronal communication. NMDARs form tetrameric complexes that consist of several homologous subunits. The subunit composition of NMDARs is plastic, resulting in a large number of receptor subtypes. As each receptor subtype has distinct biophysical, pharmacological and signalling properties, there is great interest in determining whether individual subtypes carry out specific functions in the CNS in both normal and pathological conditions. Here, we review the effects of subunit composition on NMDAR properties, synaptic plasticity and cellular mechanisms implicated in neuropsychiatric disorders. Understanding the rules and roles of NMDAR diversity could provide new therapeutic strategies against dysfunctions of glutamatergic transmission.
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            How to calculate sample size in animal studies?

            Calculation of sample size is one of the important component of design of any research including animal studies. If a researcher select less number of animals it may lead to missing of any significant difference even if it exist in population and if more number of animals selected then it may lead to unnecessary wastage of resources and may lead to ethical issues. In this article, on the basis of review of literature done by us we suggested few methods of sample size calculations for animal studies.
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              Autistic-like social behaviour in Shank2-mutant mice improved by restoring NMDA receptor function.

              Autism spectrum disorder (ASD) is a group of conditions characterized by impaired social interaction and communication, and restricted and repetitive behaviours. ASD is a highly heritable disorder involving various genetic determinants. Shank2 (also known as ProSAP1) is a multi-domain scaffolding protein and signalling adaptor enriched at excitatory neuronal synapses, and mutations in the human SHANK2 gene have recently been associated with ASD and intellectual disability. Although ASD-associated genes are being increasingly identified and studied using various approaches, including mouse genetics, further efforts are required to delineate important causal mechanisms with the potential for therapeutic application. Here we show that Shank2-mutant (Shank2(-/-)) mice carrying a mutation identical to the ASD-associated microdeletion in the human SHANK2 gene exhibit ASD-like behaviours including reduced social interaction, reduced social communication by ultrasonic vocalizations, and repetitive jumping. These mice show a marked decrease in NMDA (N-methyl-D-aspartate) glutamate receptor (NMDAR) function. Direct stimulation of NMDARs with D-cycloserine, a partial agonist of NMDARs, normalizes NMDAR function and improves social interaction in Shank2(-/-) mice. Furthermore, treatment of Shank2(-/-) mice with a positive allosteric modulator of metabotropic glutamate receptor 5 (mGluR5), which enhances NMDAR function via mGluR5 activation, also normalizes NMDAR function and markedly enhances social interaction. These results suggest that reduced NMDAR function may contribute to the development of ASD-like phenotypes in Shank2(-/-) mice, and mGluR modulation of NMDARs offers a potential strategy to treat ASD.
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                Author and article information

                Contributors
                Role: InvestigationRole: MethodologyRole: Writing – original draftRole: Writing – review & editing
                Role: InvestigationRole: Project administrationRole: SoftwareRole: Writing – original draftRole: Writing – review & editing
                Role: MethodologyRole: SoftwareRole: Visualization
                Role: Formal analysisRole: SoftwareRole: Visualization
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Funding acquisitionRole: MethodologyRole: ResourcesRole: SupervisionRole: ValidationRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS One
                plos
                PLOS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                10 October 2023
                2023
                : 18
                : 10
                : e0292631
                Affiliations
                [001] Medical Plants Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran
                Belgrade University Faculty of Medicine, SERBIA
                Author notes

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

                Author information
                https://orcid.org/0000-0003-4625-6258
                https://orcid.org/0000-0003-3210-2338
                Article
                PONE-D-23-12359
                10.1371/journal.pone.0292631
                10564128
                37815997
                d4b3a07c-e1ae-42cd-9afa-c4796c53d447
                © 2023 Khaledi 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
                : 24 April 2023
                : 25 September 2023
                Page count
                Figures: 6, Tables: 1, Pages: 17
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100005756, Shahrekord University of Medical Sciences;
                Award ID: 3533
                Award Recipient :
                This study was supported by a research grant (3533) from Shahrekord University of Medical Sciences, Shahrekord, Iran. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Psychology
                Developmental Psychology
                Pervasive Developmental Disorders
                Autism Spectrum Disorder
                Autism
                Social Sciences
                Psychology
                Developmental Psychology
                Pervasive Developmental Disorders
                Autism Spectrum Disorder
                Autism
                Medicine and Health Sciences
                Medical Conditions
                Neurodevelopmental Disorders
                Autism
                Biology and Life Sciences
                Neuroscience
                Developmental Neuroscience
                Neurodevelopmental Disorders
                Autism
                Medicine and Health Sciences
                Neurology
                Neurodevelopmental Disorders
                Autism
                Biology and Life Sciences
                Psychology
                Behavior
                Animal Behavior
                Social Sciences
                Psychology
                Behavior
                Animal Behavior
                Biology and Life Sciences
                Zoology
                Animal Behavior
                Biology and Life Sciences
                Psychology
                Developmental Psychology
                Pervasive Developmental Disorders
                Autism Spectrum Disorder
                Social Sciences
                Psychology
                Developmental Psychology
                Pervasive Developmental Disorders
                Autism Spectrum Disorder
                Biology and Life Sciences
                Anatomy
                Brain
                Hippocampus
                Medicine and Health Sciences
                Anatomy
                Brain
                Hippocampus
                Biology and Life Sciences
                Genetics
                Gene Expression
                Research and Analysis Methods
                Animal Studies
                Experimental Organism Systems
                Model Organisms
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                Research and Analysis Methods
                Model Organisms
                Mouse Models
                Research and Analysis Methods
                Animal Studies
                Experimental Organism Systems
                Animal Models
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                Physical Sciences
                Chemistry
                Chemical Compounds
                Nitrites
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Vertebrates
                Amniotes
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                Biology and Life Sciences
                Zoology
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