ABSTRACT
Under physiological conditions, the membranes and lipid droplets of germ cells are in a conformationally disordered phase. Typically, during cooling, lipids undergo the transition to ordered phases and, upon heating, melt into a disordered phase. In this communication, we report the lipid phase transition in lipid droplets observed in porcine oocytes. Upon cooling, a sharp lipid phase transition from conformationally disordered to ordered state was detected within the temperature range between 20 and 15 °C. Subsequent heating to 45 °C does not return lipids to their original phase state. To the best of our knowledge, this is the first observation of an irreversible phase transition in lipid droplets of biological cells with native lipid composition.
Subject(s)
Cryopreservation , Oocytes , Animals , Swine , Cryopreservation/methods , Phase Transition , Freezing , LipidsABSTRACT
Lipids significantly affect embryo cryopreservation in some mammalian species depending on the cell lipidome quantity and composition. One of the ways to study the relationship between lipid content and cryotolerance of cells is to study the effect of lipidome modification on laboratory mice. The objective of this research was to study how in vitro culture of mouse embryos with linoleic acid (LA) will affect lipid phase transition (LPT) during cooling and subsequent embryo development after cryopreservation. Embryos obtained in vivo at the 2-cell stage were cultured with 200 µM LA for 46 h up to the morula/blastocyst stage. Thereafter, one portion of embryos was slowly frozen to reveal the effect of LA on survival after cryopreservation, another portion was used to characterize the lipid composition and to determine the temperature of the LPT onset. Confocal fluorescence microscopy of Nile Red stained embryos showed a significant increase in lipid content of the LA treated group compared to the controls. Raman measurements showed that the onset of LPT in LA treated embryos is lower than in untreated ones: -5 °C vs +2 °C. However, these changes in the LPT onset did not affect the survival rates of embryos after cryopreservation. In summary, in vitro culture with LA changes the biophysical characteristics of embryos' lipidome and is realized in lower LPT onset, but this does not affect embryo survival after cryopreservation.
Subject(s)
Cryopreservation , Linoleic Acid , Animals , Blastocyst , Cryopreservation/methods , Freezing , Linoleic Acid/pharmacology , Lipids , MiceABSTRACT
There are evidences that obese women exhibit a detrimental oocyte quality. However, it remains unclear how this change is associated with obesity, indirectly - or directly through a change in the content and/or composition of lipids in oocytes. The aim of this work was to study effects of a high-fat diet applied to female donor mice on the amount and qualitative composition of lipids of immature and in vivo matured oocytes. A high-fat diet caused larger body weight in female mice compared with the control ( p < 0.001; 44.77 ± 1.46 and 35.22 ± 1.57, respectively), and increased the blood levels of cholesterol ( p < 0.05; 2.06 ± 0.10 and 1.78 ± 0.10, respectively) and triglycerides ( p < 0.05; 2.13 ± 0.23 and 1.49 ± 0.21, respectively). At the same time, this diet does not affect the level of unsaturation of lipids in immature (0.207 ± 0.004 in the experiment and 0.206 ± 0.002 in the control) and matured oocytes (0.212 ± 0.005 in the experiment and 0.211 ± 0.003 in the control). Total lipid content increased during in vivo maturation of mouse oocytes. The amount of lipids was greater in mature oocytes in the experimental group compared to the control ( p < 0.01; 8.15 ± 0.37 and 5.83 ± 0.14, respectively). An increase in intracellular lipid amount during oocyte maturation was revealed both after a standard diet ( p < 0.05; 4.72 ± 0.48 and 5.83 ± 0.14, respectively) and after a fat-rich diet ( p < 0.001; 3.45 ± 0.62 and 8.15 ± 0.37, respectively). Thus, during in vivo oocyte maturation in mice the content of intracellular lipids enhanced, the high-fat diet aggravated this dynamics of lipid increase during in vivo maturation of oocytes.
