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S'acclimater À Des Salinités Et Températures Changeantes: Un Focus Sur La Branchie Chez Le Loup Européen Dicentrarchus L

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S'acclimater À Des Salinités Et Températures Changeantes: Un Focus Sur La Branchie Chez Le Loup Européen Dicentrarchus L S’acclimater à des salinités et températures changeantes : un focus sur la branchie chez le loup européen Dicentrarchus labrax Waliullah Masroor To cite this version: Waliullah Masroor. S’acclimater à des salinités et températures changeantes : un focus sur la branchie chez le loup européen Dicentrarchus labrax. Agricultural sciences. Université Montpellier, 2019. English. NNT : 2019MONTG008. tel-02142198 HAL Id: tel-02142198 https://tel.archives-ouvertes.fr/tel-02142198 Submitted on 28 May 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. THÈSE POUR OBTENIR LE GRADE DE DOCTEUR DE L’UNIVERSITÉ DE MONTPELLIER En Ecophysiologie Adaptative École doctorale GAIA Biodiversité, Agriculture, Alimentation, Environnement, Terre, Eau Unité de recherche MARBEC Marine Biodiversity, Exploitation and Conservation COPING WITH SALINITY AND TEMPERATURE CHANGES: A FOCUS ON THE GILL RESPONSE IN EUROPEAN SEA BASS Dicentrarchus labrax Présentée par Waliullah MASROOR Le 26 Avril 2019 Sous la direction Mme Catherine LORIN-NEBEL et Mme Emilie FARCY Devant le jury composé de Mme Delphine DESTOUMIEUX-GARZON, Directrice de Recherche, CNRS, Université Montpellier Présidente M. Gert FLIK, Professeur, Radboud University, Pays Bas Rapporteur Mme Anne-Sophie MARTINEZ, Maître de Conférences, Université de Caen Normandie Rapporteur M. Jack FALCON, Directeur de Recherche Emerite, CNRS, Université Pierre et Marie Curie Examinateur Mme Emilie FARCY, Maître de Conférences, Université de Montpellier Co-encadrante Mme Catherine LORIN-NEBEL, Maître de Conférences, Université de Montpellier Directrice THÈSE POUR OBTENIR LE GRADE DE DOCTEUR DE L’UNIVERSITÉ DE MONTPELLIER En Ecophysiologie Adaptative École doctorale GAIA Biodiversité, Agriculture, Alimentation, Environnement, Terre, Eau Unité de recherche MARBEC Marine Biodiversity, Exploitation and Conservation COPING WITH SALINITY AND TEMPERATURE CHANGES: A FOCUS ON THE GILL RESPONSE IN EUROPEAN SEA BASS Dicentrarchus labrax Présentée par Waliullah MASROOR Le 26 Avril 2019 Sous la direction Mme Catherine LORIN-NEBEL et Mme Emilie FARCY Devant le jury composé de Mme Delphine DESTOUMIEUX-GARZON, Directrice de Recherche, CNRS, Université Montpellier Présidente M. Gert FLIK, Professeur, Radboud University, Pays Bas Rapporteur Mme Anne-Sophie MARTINEZ, Maître de Conférences, Université de Caen Normandie Rapporteur M. Jack FALCON, Directeur de Recherche Emerite, CNRS, Université Pierre et Marie Curie Examinateur Mme Emilie FARCY, Maître de Conférences, Université de Montpellier Co-encadrante Mme Catherine LORIN-NEBEL, Maître de Conférences, Université de Montpellier Directrice Dedication Dedication I dedicate this thesis to my beloved younger brother Abdul Muqeet Umair (late) (2007-2018) not to see him in my family any more. I never thought about this day that upon completion of my thesis I won’t be able to see him celebrating my PhD. i Acknowledgement Acknowledgement Many people have inspired and supported me for completion of this thesis, but to the greatest extent, I wish to express my heartfelt gratitude to my supervisors Dr. Catherine Lorin-Nebel and Dr. Emilie Farcy. I will be indebted to them that they reshaped my academic career by accepting me as a PhD student and this means a lot to me. It is merely possible because of their encouragement and guidance this thesis never would have eventuated. Thank you for giving me this opportunity, never letting me give up, and always finding time for me. Also special thanks to them they teach me many more things that could have been learnt before starting my PhD. I remember when I started my PhD, it was difficult for me to find myself and I was always lost then it were my supervisors their feedback and troubleshooting made my way to move forward. They deserved more than words and I truly appreciate for their corrections and presentation skills they taught me. I wish to thank the jury members of my thesis: Gert Flik, Anne-Sophie Martinez, Jack Falcon and Delphine Destoumieux for generously offer their time and review of this document. I admire magnificent suggestions of thesis committee members: Helena D’Cotta, Delphine Destoumieux, Benjamin Geffroy and Julien Issartel that improved my research work and they endorsed my efforts of each year. I submit my heartiest gratitude to Prof. Guy Charmantier for his guidance in my experiments. I am deeply indebted to my other lab members: Jehan-Herve Lignot, Patricia Cucchi, Eva Blondeau-Bidet, Sophie Hermet, Romain Gros and colleagues: Thibaut, Quanquan, Khalid, Elsa, Laura, Thomas, Dimitri for their invaluable help in the lab and sometime outside the lab when it comes to speak French. My country fellows were nice and supportive at all stages I humbly extend my thanks to all. To my loving parents, sisters and brothers, I offer my sincerest thanks for their unswerving support for my stay in France and of my work and for the patience and understanding that they have shown despite me having far too much time away from them. With the thesis now finished, it’s finally time for us to meet in home country. I look forward to all the exciting times that lie ahead. My joy knows no bounds in expressing my cordial gratitude to my loving wife and daughter. They were always by my side and served my inspiration to complete my studies. I would like to express my gratitude for being with me in France and my wife’s interest in baking bakery items were the great part I enjoyed most. iii Table of contents Table of Contents 1 Chapter I – Introduction…………………………………………………………….....3 1.1 Environmental variability in Mediterranean coastal areas: present and future..3 1.1.1 Characterization of West Mediterranean lagoons and estuaries…….....3 1.2 Future: what modifications are expected in the context of climate change? ….4 1.2.1 Climate change – global context……………………..……………...…4 1.2.2 Climate change impacts on Mediterranean Sea…………………..……6 1.2.3 Climate change impacts on coastal estuaries and lagoons habitats……6 1.3 Climate change impacts on aquatic animals………………………………...…9 1.3.1 Biodiversity and species extinction………………………..………..…9 1.3.2 Habitat distribution………………………..………………………...…9 1.3.3 Thermal performance …………………………………..…..………..10 1.3.4 Acidification and hypoxia………………………………..…………..10 1.4 Environmental stress and fish acclimation…………………………..…...…..11 1.4.1 Temperature acclimation…………………………………..………....11 1.4.2 Salinity acclimation…………………………………..………..……..15 1.4.3 Acid base regulation…………………………………………...……..19 1.4.4 Combined effects of temperature and salinity………………………..20 1.4.5 Objectives………………………………………..……………….…..23 2 Chapter II - Materials and Methods……………………………………………….....27 2.1 European sea bass Dicentrarchus labrax………………………………….…27 2.2 Life cycle of sea bass ……………………………………………..……...….27 2.3 Sea bass as an experimental fish………………………………………….….28 2.4 Experimental conditions………………………………..…………………….28 2.5 Blood osmolality………………………………..….………………….……...30 2.6 Sodium ion levels…………………………………………..………….……...30 2.7 Chloride ion levels………………………………………………….….……..30 2.8 Light microscopy and immunofluorescence analysis……….……………......30 2.8.1 Tissue sectioning………………………..…………………..….……..30 2.8.2 Gill staining……………………………………...………….….……..31 2.8.3 Morphometric measurement………………...………….……………..31 2.8.4 Immunofluorescence observation………………...…….……………..31 2.8.5 Counting………………………...…………………….………………32 2.8.6 Ionocytes area measurements…………………...….………………....32 2.9 Scanning electron microscopy….…………………………….……………….32 2.10 Gill Na+/K+-ATPase activity measurement………………………..……..…..33 2.11 RNA extraction purification and reverse transcription………………….…....33 2.12 Quantification at the transcript level………………………………….……....34 3 Chapter III - Coping with environmental changes: from the cell to the whole organism.. ………………………...…………………….………………….…39 3.1 Preamble…………………………………………………………………........39 3.2 Main findings……………………………………………………………...….40 3.3 Conclusion………………………………………………………………….....42 Research article 1 - Effect of salinity and temperature in sea bass osmoregulatory processes…………………………………...……………………………………….....45 v Table of contents 4 Chapter IV - Coping with environmental changes: gene expression……..…………..57 4.1 Preamble……………………………………………………………...……....57 4.2 Main findings…………………………………………………………...…....58 4.3 Conclusion…..………………………………………………………………..60 Research article 2 - Effects on branchial ion transports: gene expression………..…..63 5 Chapter V - Temperature and salinity stress affect gill mucus and heat stress-related proteins………………………………………………………………………………..91 5.1 Preamble……………………………………………………………...……….91 5.2 Major highlights……………………………………………………………....92 Research article 3 - Temperature and salinity stress affect gill mucus and heat stress-related proteinsfects on mucus and heat stress-related proteins………………..95 6 Chapter VI - General discussion………………………………………………….....113 6.1 Effects of combined stress on physiological, biochemical and morphological parameters……………...................................................................................117 6.2 Temperature stress modulates branchial mRNA levels of
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