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      Transient Receptor Potential-Vanilloid (TRPV1-TRPV4) Channels in the Atlantic Salmon, Salmo salar. A Focus on the Pineal Gland and Melatonin Production

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          Abstract

          Fish are ectotherm, which rely on the external temperature to regulate their internal body temperature, although some may perform partial endothermy. Together with photoperiod, temperature oscillations, contribute to synchronizing the daily and seasonal variations of fish metabolism, physiology and behavior. Recent studies are shedding light on the mechanisms of temperature sensing and behavioral thermoregulation in fish. In particular, the role of some members of the transient receptor potential channels (TRP) is being gradually unraveled. The present study in the migratory Atlantic salmon, Salmo salar, aims at identifying the tissue distribution and abundance in mRNA corresponding to the TRP of the vanilloid subfamilies, TRPV1 and TRPV4, and at characterizing their putative role in the control of the temperature-dependent modulation of melatonin production—the time-keeping hormone—by the pineal gland. In Salmo salar, TRPV1 and TRPV4 mRNA tissue distribution appeared ubiquitous; mRNA abundance varied as a function of the month investigated. In situ hybridization and immunohistochemistry indicated specific labeling located in the photoreceptor cells of the pineal gland and the retina. Additionally, TRPV analogs modulated the production of melatonin by isolated pineal glands in culture. The TRPV1 agonist induced an inhibitory response at high concentrations, while evoking a bell-shaped response (stimulatory at low, and inhibitory at high, concentrations) when added with an antagonist. The TRPV4 agonist was stimulatory at the highest concentration used. Altogether, the present results agree with the known widespread distribution and role of TRPV1 and TRPV4 channels, and with published data on trout ( Oncorhynchus mykiss), leading to suggest these channels mediate the effects of temperature on S. salar pineal melatonin production. We discuss their involvement in controlling the timing of daily and seasonal events in this migratory species, in the context of an increasing warming of water temperatures.

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
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            A new mathematical model for relative quantification in real-time RT-PCR.

            M. Pfaffl (2001)
            Use of the real-time polymerase chain reaction (PCR) to amplify cDNA products reverse transcribed from mRNA is on the way to becoming a routine tool in molecular biology to study low abundance gene expression. Real-time PCR is easy to perform, provides the necessary accuracy and produces reliable as well as rapid quantification results. But accurate quantification of nucleic acids requires a reproducible methodology and an adequate mathematical model for data analysis. This study enters into the particular topics of the relative quantification in real-time RT-PCR of a target gene transcript in comparison to a reference gene transcript. Therefore, a new mathematical model is presented. The relative expression ratio is calculated only from the real-time PCR efficiencies and the crossing point deviation of an unknown sample versus a control. This model needs no calibration curve. Control levels were included in the model to standardise each reaction run with respect to RNA integrity, sample loading and inter-PCR variations. High accuracy and reproducibility (<2.5% variation) were reached in LightCycler PCR using the established mathematical model.
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              ThermoTRP channels and beyond: mechanisms of temperature sensation.

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                Author and article information

                Contributors
                Journal
                Front Physiol
                Front Physiol
                Front. Physiol.
                Frontiers in Physiology
                Frontiers Media S.A.
                1664-042X
                07 January 2022
                2021
                : 12
                : 784416
                Affiliations
                [1] 1Sorbonne Université (SU), CNRS, Biologie Intégrative des Organismes Marins (BIOM) , Banyuls-sur-Mer, France
                [2] 2Conservatoire National du Saumon Sauvage , Chanteuges, France
                [3] 3SU, CNRS Fédération 3724, Observatoire Océanologique , Banyuls-sur-Mer, France
                [4] 4Departamento de Genética, Fisiología y Microbiologia, Facultad de Biología, Universidad Complutense de Madrid , Madrid, Spain
                Author notes

                Edited by: José A. Muñoz-Cueto, University of Cádiz, Spain

                Reviewed by: Sebastian Boltana, University of Concepcion, Chile; Francesc Piferrer, Institute of Marine Sciences, Spanish National Research Council (CSIC), Spain

                *Correspondence: Laura Gabriela Nisembaum, laugabri10@ 123456gmail.com

                Present address: Guillaume Loentgen, Marine Biology Department, Centre Scientifique de Monaco, Monaco, Monaco; Thibaut L’Honoré, Centre Universitaire de Formation et de Recherche de Mayotte (CUFR), Mayotte, France; Charles-Hubert Paulin, SU, CNRS Fédération 3724, Observatoire Océanologique, Banyuls-sur-Mer, France; Jack Falcón, Muséum National d’Histoire Naturelle (MNHN), Biologie des Organismes et Ecosystèmes Aquatiques (BOREA) CNRS UMR 8067, SU, IRD 207, UCN, UA, Paris, France

                This article was submitted to Aquatic Physiology, a section of the journal Frontiers in Physiology

                Article
                10.3389/fphys.2021.784416
                8782258
                35069244
                bdcd75bb-cc4d-4596-b203-b252cc9545af
                Copyright © 2022 Nisembaum, Loentgen, L’Honoré, Martin, Paulin, Fuentès, Escoubeyrou, Delgado, Besseau and Falcón.

                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
                : 27 September 2021
                : 07 December 2021
                Page count
                Figures: 6, Tables: 1, Equations: 0, References: 94, Pages: 15, Words: 10650
                Funding
                Funded by: Agence Nationale de la Recherche , doi 10.13039/501100001665;
                Funded by: Conseil National de la Recherche Scientifique , doi 10.13039/501100007175;
                Funded by: Sorbonne Université , doi 10.13039/501100019125;
                Categories
                Physiology
                Original Research

                Anatomy & Physiology
                atlantic salmon,temperature,pineal organ,melatonin,transient receptor potential vanilloid (trpv),trpv1,trpv4

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