Time-resolved Fluorescence and Generalized Polarization: Innovative tools to assess bull sperm membrane dynamics during slow freezing

During slow freezing, spermatozoa undergo membrane alterations that compromise their ability of fertilizing. These alterations are cause either by cold shock or by the use of cryoprotectants known to be cytotoxic. However, little is known about the membrane changes that occurred during freezing. Her...

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Bibliographic Details
Published in:Cryobiology 2019-12, Vol.91, p.69-76
Main Authors: Bechoua, Shaliha, Winckler, Pascale, Jossier, Audrey, Peltier, Caroline, Delize, Frédéric, Devaux, Noémie, Perrier Cornet, Jean Marie, Simonin, Hélène
Format: Article
Language:English
Subjects:
2-Naphthylamine - analogs & derivatives
2-Naphthylamine - chemistry
Animals
Bull sperm
Cattle
Cell Membrane - physiology
Cryopreservation - methods
Cryoprotective Agents - pharmacology
Engineering Sciences
Fluorescence
Freezing
Laurates - chemistry
Male
Membrane
Membrane Fluidity - physiology
Slow freezing
Sperm Motility - physiology
Spermatozoa - physiology
Time-resolved fluorescence
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Summary:During slow freezing, spermatozoa undergo membrane alterations that compromise their ability of fertilizing. These alterations are cause either by cold shock or by the use of cryoprotectants known to be cytotoxic. However, little is known about the membrane changes that occurred during freezing. Here, we combined Generalized Polarization (GP), Time-resolved Fluorescence and laurdan fluorescence properties to investigate the changes in membrane fluidity and dynamics during slow freezing of bull sperm. We successfully demonstrated that laurdan may be distributed in three different local environments that correspond to different membrane lipid composition. These environments wont behave the same way when the cells will be subjected to either a chemical treatment (adding the cryoprotectants) or a physical treatment (freezing).
ISSN:0011-2240
1090-2392
DOI:10.1016/j.cryobiol.2019.10.196