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      Ghrelin misbalance affects mice embryo implantation and pregnancy success by uterine immune dysregulation and nitrosative stress

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          Abstract

          Introduction

          In a previous study we found that ghrelin (Ghrl) misbalance during the peri-implantation period significantly impaired fetus development. In this study we aimed to evaluate the putative mechanisms underlying these effects, including embryo implantation success, uterine nitric oxide synthase (NOS) activity, nitric oxide synthesis and the inflammatory/immune uterine profile.

          Methods

          Ghrelin misbalance was induced by injecting 4nmol/animal/day of Ghrl (hyperghrelinemia) or 6nmol/animal/day of a Ghrl antagonist (Ant: (D-Lys3)GHRP-6) from day 3 to 8 of pregnancy. Control animals (C) were injected with de vehicle. Females were euthanized at pregnancy day 8 and their uteri excised in order to evaluate: the percentage of reabsorbed embryos (microscopically), eNOS, iNOS and nytrotirosine expression (by immunohistochemistry), nitrite synthesis (by Griess technique), VEGF, IL-10, IL-17, IL-6, MMP9 and GM-CSF expression (by qPCR) and leukocyte infiltration by flow cytometry (evaluating T cells, NK cells, granulocytes, dendritic cells and macrophages).

          Results

          Ant-treatment significantly increased the percentage of reabsorbed embryos and the uterine expression of eNOS, iNOS and nytrotirosine. (D-Lys3)GHRP-6-treatment increased also the expression of the inflammatory cytokines IL-6, IL-17 and MMP9, and decreased that of IL-10 (anti-inflammatory). Moreover, Ant-treatment increased also the NK cells population and that of CD11b + dendritic cells; and decreased T cells percentages. Similarly, hyperghrelinemia showed a significant increase vs. C on eNOS, iNOS and nytrotirosineuterine expression and a decrease in T cells percentages.

          Conclusion

          Ghrl misbalance during the peri-implantation period induces pro-inflammatory changes and nitrosative stress in the gravid uterus, impairing significantly embryo implantation and/or development.

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          Most cited references77

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          Fiji: an open-source platform for biological-image analysis.

          Fiji is a distribution of the popular open-source software ImageJ focused on biological-image analysis. Fiji uses modern software engineering practices to combine powerful software libraries with a broad range of scripting languages to enable rapid prototyping of image-processing algorithms. Fiji facilitates the transformation of new algorithms into ImageJ plugins that can be shared with end users through an integrated update system. We propose Fiji as a platform for productive collaboration between computer science and biology research communities.
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            Ghrelin is a growth-hormone-releasing acylated peptide from stomach.

            Small synthetic molecules called growth-hormone secretagogues (GHSs) stimulate the release of growth hormone (GH) from the pituitary. They act through GHS-R, a G-protein-coupled receptor for which the ligand is unknown. Recent cloning of GHS-R strongly suggests that an endogenous ligand for the receptor does exist and that there is a mechanism for regulating GH release that is distinct from its regulation by hypothalamic growth-hormone-releasing hormone (GHRH). We now report the purification and identification in rat stomach of an endogenous ligand specific for GHS-R. The purified ligand is a peptide of 28 amino acids, in which the serine 3 residue is n-octanoylated. The acylated peptide specifically releases GH both in vivo and in vitro, and O-n-octanoylation at serine 3 is essential for the activity. We designate the GH-releasing peptide 'ghrelin' (ghre is the Proto-Indo-European root of the word 'grow'). Human ghrelin is homologous to rat ghrelin apart from two amino acids. The occurrence of ghrelin in both rat and human indicates that GH release from the pituitary may be regulated not only by hypothalamic GHRH, but also by ghrelin.
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              The dendritic cell lineage: ontogeny and function of dendritic cells and their subsets in the steady state and the inflamed setting.

              Dendritic cells (DCs) form a remarkable cellular network that shapes adaptive immune responses according to peripheral cues. After four decades of research, we now know that DCs arise from a hematopoietic lineage distinct from other leukocytes, establishing the DC system as a unique hematopoietic branch. Recent work has also established that tissue DCs consist of developmentally and functionally distinct subsets that differentially regulate T lymphocyte function. This review discusses major advances in our understanding of the regulation of DC lineage commitment, differentiation, diversification, and function in situ.
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                Author and article information

                Contributors
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                URI : https://loop.frontiersin.org/people/532157Role: Role: Role: Role: Role:
                URI : https://loop.frontiersin.org/people/765219Role: Role: Role: Role:
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                URI : https://loop.frontiersin.org/people/83446Role: Role: Role: Role: Role: Role: Role:
                URI : https://loop.frontiersin.org/people/1646525Role: Role: Role: Role: Role: Role: Role: Role: Role:
                Journal
                Front Endocrinol (Lausanne)
                Front Endocrinol (Lausanne)
                Front. Endocrinol.
                Frontiers in Endocrinology
                Frontiers Media S.A.
                1664-2392
                01 December 2023
                2023
                : 14
                : 1288779
                Affiliations
                [1] 1 Instituto de Fisiología, Facultad de Ciencias Médicas, Universidad Nacional de Córdoba , Córdoba, Argentina
                [2] 2 Instituto de Investigaciones en Ciencias de la Salud (INICSA), Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET)/Universidad Nacional de Córdoba (UNC) , Córdoba, Argentina
                [3] 3 Instituto de Biología Celular, Facultad de Ciencias Médicas, Universidad Nacional de Córdoba , Córdoba, Argentina
                [4] 4 Departamento de Bioquímica Clínica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba , Córdoba, Argentina
                [5] 5 Centro de Investigaciones en Bioquímica Clínica e Inmunología (CIBICI), Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET)/Universidad Nacional de Córdoba (UNC) , Córdoba, Argentina
                Author notes

                Edited by: Maria Garces, National University of Colombia, Colombia

                Reviewed by: Anne Schumacher, Helmholtz Association of German Research Centres (HZ), Germany; Estela Bevilacqua, University of São Paulo, Brazil; Hannah Ee Juen Yong, Singapore Institute for Clinical Sciences (A*STAR), Singapore

                *Correspondence: Ana Carolina Martini, acmartini2000@ 123456fcm.unc.edu.ar

                †These authors have contributed equally to this work and share last authorship

                Article
                10.3389/fendo.2023.1288779
                10722256
                38107518
                3a4341a8-76be-48e4-8ce7-8ed49ccc9d0c
                Copyright © 2023 Luque, Díaz-Luján, Paira, de Loredo, Torres, Cantarelli, Fretes, Motrich and Martini

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 04 September 2023
                : 09 November 2023
                Page count
                Figures: 4, Tables: 1, Equations: 0, References: 78, Pages: 12, Words: 6568
                Funding
                The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This study was supported by grants from the Secretaría de Ciencia y Tecnología, Universidad Nacional de Córdoba (SECyT-UNC, 2018-2022), Fondo para la Investigación Científica y Tecnológica (PICT2015-1847, PICT2019-2451), and Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET; PIP 2014-2016).
                Categories
                Endocrinology
                Original Research
                Custom metadata
                Reproduction

                Endocrinology & Diabetes
                (d-lys3)ghrp-6,nos,nitrosative stress,ils,mmp9,nk cells,dendritic cells,t cells
                Endocrinology & Diabetes
                (d-lys3)ghrp-6, nos, nitrosative stress, ils, mmp9, nk cells, dendritic cells, t cells

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