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ROLE OF IONS IN THE SPERM ACTIVATION after activation in eel sperm, as is the case with pufferfish. Also, our results agree with those of Boitano & Omoto (1991), regarding rainbow trout sperm, where an intracellular acidification upon sperm hypoosmotic activation was observed. These authors considered that changes in intracellular pH do not regulate trout sperm motility, contrarily to what happens in other species. The slow decrease in pH over time observed in their study suggests that this change could be a consequence of motility activation, rather than a trigger for sperm motility initiation. The observed pH i decrease might simply be the result of the mitochondrial sperm respiration during the active movement. Finally, regarding the ion distribution in quiescent European eel spermatozoa, fluorescence images showed that Ca 2+ and K + were located mainly in the mitochondrion, which in this species is a single, small and round organelle, located in the apex of the spermatozoa head, opposite the axoneme (Marco-Jiménez et al., 2006). The accumulation of calcium stores in the mitochondrion has previously been observed in human sperm (Costello et al., 2009), but potassium accumulation has not been reported in any other species studied so far. However, the existence of specific potassium channels in the mitochondria of several animal tissues (liver, heart, brain, kidney, skeletal muscle, human T lymphocytes and amoeba; review in Szewczyk et al., 2009) indicates that a differential concentration of K + between this organelle and the cytoplasm is possible in these tissues. It is not known whether sperm mitochondria possess such K + channels, but the accumulation of the ion in this organelle therefore suggests an important role of the eel sperm mitochondria in the function of the sperm. In conclusion, intracellular concentrations of Ca 2+ and K + ions in sperm increase after hyperosmotic sperm activation in European eel, with a progressive decrease in intracellular pH. Fluorescence images suggest an accumulation of both ions in the mitochondrion, which could represent ion stores. However, further studies including the use of ion channels blockers, activation media with/without specific ions, as well as changes in cell volume, may be necessary to determine the fluxes of these ions and their role in motility initiation on the European eel sperm. 81
CHAPTER 3 Acknowledgements Funded from the European Community's 7th Framework Programme under the Theme 2 "Food, Agriculture and Fisheries, and Biotechnology", grant agreement n°245257 (Pro-Eel) and the Spanish Ministry of Science and Innovation (MICINN; AGL2010-16009). Victor Gallego has a predoctoral grant (MICINN; BES-2009-020310) and has been granted a fellowship (EEBB- I-12-05858) of the Spanish Personnel Research Training Programme to carry out this study in the Universidad de León (León, Spain). Ilaria Mazzeo had a predoctoral grant from GVA. David S. Peñaranda has a contract co-financed by MICINN and UPV (PTA2011-4948-I). F. Martínez-Pastor was supported by the Ramón y Cajal program (RYC-2008-02560, MICINN). 82
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SPERM PHYSIOLOGY AND QUALITY IN TWO
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ACKNOWLEDGEMENTS Muchos años han p
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aunque tiene una alcachofa en la ca
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TABLE OF CONTENTS SUMMARY 1 RESUMEN
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SUMMARY SUMMARY The conservation st
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RESUMEN RESUMEN Las especies objeto
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RESUM RESUM Les espècies objecte d
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GENERAL INTRODUCTION 7
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GENERAL INTRODUCTION who postulated
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GENERAL INTRODUCTION 2. Sperm physi
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GENERAL INTRODUCTION Finally, the a
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GENERAL INTRODUCTION 3. Sperm quali
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GENERAL INTRODUCTION in fish (Kime
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GENERAL INTRODUCTION However, for s
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GENERAL INTRODUCTION future applica
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GENERAL INTRODUCTION With regards t
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OBJECTIVES The chapters presented b
Page 40 and 41: STANDARDIZATION OF SPERM MOTILITY E
Page 56: CHAPTER 2 Study of the effects of t
Page 59 and 60: CHAPTER 2 reducing the pressure on
Page 61 and 62: CHAPTER 2 ongoing task in order to
Page 63 and 64: CHAPTER 2 To measure sperm density,
Page 65 and 66: CHAPTER 2 Spermiating males (%) 100
Page 67 and 68: CHAPTER 2 100 80 60 I II III IV A 4
Page 69 and 70: CHAPTER 2 3.2 Hormonal treatments P
Page 71 and 72: CHAPTER 2 With regard to the sperm
Page 73 and 74: CHAPTER 2 Relative volume (%) 80 I
Page 75 and 76: CHAPTER 2 In this respect, it is we
Page 77 and 78: CHAPTER 2 treatment to that of the
Page 79 and 80: CHAPTER 2 Our results demonstrate t
Page 82 and 83: ROLE OF IONS IN THE SPERM ACTIVATIO
Page 92: CHAPTER 4 Study of pufferfish (Taki
Page 95 and 96: CHAPTER 4 2002) so Takifugu niphobl
Page 97 and 98: CHAPTER 4 carrying out sperm collec
Page 99 and 100: CHAPTER 4 Table 1. Activation media
Page 101 and 102: CHAPTER 4 TM (%) 100 A a a Und-PD a
Page 103 and 104: CHAPTER 4 TM (%) 80 60 40 20 0 A a
Page 105 and 106: CHAPTER 4 TM (%) 80 A ASW100 GLU 60
Page 107 and 108: CHAPTER 4 although the activation m
Page 109 and 110: CHAPTER 4 dilution ratio and an opt
Page 111 and 112: CHAPTER 4 2006). Several studies in
Page 113 and 114: CHAPTER 4 5. Conclusions Some concl
Page 116 and 117: RELATIONSHIP BETWEEN SPERM MOTILIY
Page 130: RELATIONSHIP BETWEEN SPERM MOTILIY
Page 134 and 135: GENERAL DISCUSSION 1. Improvements
Page 136 and 137: GENERAL DISCUSSION profiles induced
Page 138 and 139: GENERAL DISCUSSION more studies on
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GENERAL DISCUSSION sperm motility p
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CONCLUSIONS 133
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CONCLUSIONS vii. The coefficients o
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REFERENCES Aarestrup K, Økland F,
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REFERENCES Beirão J, Cabrita E, P
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REFERENCES Cabrita E, Robles V, Cu
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REFERENCES De Vos A, Van De Velde H
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REFERENCES Gallego V, Pérez L, Ast
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REFERENCES sperm competition and fe
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REFERENCES Liu QH, Li J, Xiao ZZ, D
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REFERENCES Mortimer ST. Effect of i
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REFERENCES Peñaranda DS, Marco-Jim
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REFERENCES Rurangwa E, Volckaert FA
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REFERENCES Tesch F. Telemetric obse
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REFERENCES Wong PYD, Lee WM, Tsang
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