Membrane permeability parameters for freezing of stallion sperm as determined by Fourier transform infrared spectroscopy.
Abstract: Cellular membranes are one of the primary sites of injury during freezing and thawing for cryopreservation of cells. Fourier transform infrared spectroscopy (FTIR) was used to monitor membrane phase behavior and ice formation during freezing of stallion sperm. At high subzero ice nucleation temperatures which result in cellular dehydration, membranes undergo a profound transition to a highly ordered gel phase. By contrast, low subzero nucleation temperatures, that are likely to result in intracellular ice formation, leave membrane lipids in a relatively hydrated fluid state. The extent of freezing-induced membrane dehydration was found to be dependent on the ice nucleation temperature, and showed Arrhenius behavior. The presence of glycerol did not prevent the freezing-induced membrane phase transition, but membrane dehydration occurred more gradual and over a wider temperature range. We describe a method to determine membrane hydraulic permeability parameters (E(Lp), Lpg) at subzero temperatures from membrane phase behavior data. In order to do this, it was assumed that the measured freezing-induced shift in wavenumber position of the symmetric CH(2) stretching band arising from the lipid acyl chains is proportional to cellular dehydration. Membrane permeability parameters were also determined by analyzing the H(2)O-bending and -libration combination band, which yielded higher values for both E(Lp) and Lpg as compared to lipid band analysis. These differences likely reflect differences between transport of free and membrane-bound water. FTIR allows for direct assessment of membrane properties at subzero temperatures in intact cells. The derived biophysical membrane parameters are dependent on intrinsic cell properties as well as freezing extender composition.
(c) 2010 Elsevier Inc. All rights reserved.
Publication Date: 2010-06-08 PubMed ID: 20553897DOI: 10.1016/j.cryobiol.2010.06.002Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article discusses how Fourier transform infrared spectroscopy (FTIR) was used to study membrane behavior and ice formation in freezing stallion sperm. The findings reveal that freezing-induced membrane dehydration depends on the ice nucleation temperature. The presence of glycerol failed to prevent this phase transition, although it made the dehydration process more gradual. The researchers also developed a method to calculate membrane permeability parameters at subzero temperatures.
Objectives of the Study
- The main aim of the study was to understand how cellular membranes, a significant site of injury in the cryopreservation of cells, react when stallion sperm is frozen.
- The authors sought to monitor the behaviour of membranes and ice formation during this freezing process by using Fourier transform infrared spectroscopy (FTIR).
- The study also aimed to develop a method for determining membrane permeability parameters at subzero temperatures.
Methodology
- To observe the phase behaviour of membranes and ice formation during the freezing of stallion sperm, the researchers employed the use of FTIR.
- Freezing-induced membrane dehydration, a significant consequence of the freezing process in cellular preservation, was studied in relation to ice nucleation temperature.
- The presence of glycerol during the freezing process was also factored in, to determine its effect on freezing-induced membrane phase transition.
- To extract membrane hydraulic permeability parameters (E(Lp), Lpg) at subzero temperatures, a new method reliant on data concerning membrane phase behavior was established.
Findings and Conclusions
- During the freezing process, high subzero ice nucleation temperatures resulted in cellular dehydration and caused the membranes to transition to a highly ordered gel phase.
- Lower subzero nucleation temperatures left the membrane lipids in a relatively hydrated fluid state, likely triggering intracellular ice formation.
- The extent of freezing-induced membrane dehydration depended largely on the ice nucleation temperature and followed Arrhenius behavior.
- Though the presence of glycerol did not halt the freezing-induced membrane phase transition, it caused membrane dehydration to occur slower and over a broader temperature range.
- The devised methodology enabled the determination of membrane permeability parameters, shedding light on the transport differences between free and membrane-bound water.
Cite This Article
APA
Oldenhof H, Friedel K, Sieme H, Glasmacher B, Wolkers WF.
(2010).
Membrane permeability parameters for freezing of stallion sperm as determined by Fourier transform infrared spectroscopy.
Cryobiology, 61(1), 115-122.
https://doi.org/10.1016/j.cryobiol.2010.06.002 Publication
Researcher Affiliations
- Clinic for Horses - Unit for Reproductive Medicine, University of Veterinary Medicine Hannover, Germany.
MeSH Terms
- Animals
- Cell Membrane / chemistry
- Cell Membrane / drug effects
- Cell Membrane Permeability / drug effects
- Cell Membrane Permeability / physiology
- Cryopreservation / methods
- Cryoprotective Agents / pharmacology
- Dehydration / metabolism
- Freezing
- Glycerol / pharmacology
- Horses
- Male
- Phase Transition / drug effects
- Semen Preservation / methods
- Spectroscopy, Fourier Transform Infrared
Citations
This article has been cited 10 times.- Kalwar Q, Chu M, Korejo RA, Soomro H, Yan P. Cryopreservation of Yak Semen: A Comprehensive Review.. Animals (Basel) 2022 Dec 7;12(24).
- Wolkers WF, Oldenhof H. Principles Underlying Cryopreservation and Freeze-Drying of Cells and Tissues.. Methods Mol Biol 2021;2180:3-25.
- Daily MI, Whale TF, Partanen R, Harrison AD, Kilbride P, Lamb S, Morris GJ, Picton HM, Murray BJ. Cryopreservation of primary cultures of mammalian somatic cells in 96-well plates benefits from control of ice nucleation.. Cryobiology 2020 Apr;93:62-69.
- Ugur MR, Saber Abdelrahman A, Evans HC, Gilmore AA, Hitit M, Arifiantini RI, Purwantara B, Kaya A, Memili E. Advances in Cryopreservation of Bull Sperm.. Front Vet Sci 2019;6:268.
- Okotrub KA, Mokrousova VI, Amstislavsky SY, Surovtsev NV. Lipid Droplet Phase Transition in Freezing Cat Embryos and Oocytes Probed by Raman Spectroscopy.. Biophys J 2018 Aug 7;115(3):577-587.
- Mazzeo DE, Marin-Morales MA. Genotoxicity evaluation of environmental pollutants using analysis of nucleolar alterations.. Environ Sci Pollut Res Int 2015 Jul;22(13):9796-806.
- Massie I, Selden C, Hodgson H, Fuller B, Gibbons S, Morris GJ. GMP cryopreservation of large volumes of cells for regenerative medicine: active control of the freezing process.. Tissue Eng Part C Methods 2014 Sep;20(9):693-702.
- Gray JE, Starmer J, Lin VS, Dickinson BC, Magnuson T. Mitochondrial hydrogen peroxide and defective cholesterol efflux prevent in vitro fertilization by cryopreserved inbred mouse sperm.. Biol Reprod 2013 Jul;89(1):17.
- Lin CY, Wei PL, Chang WJ, Huang YK, Feng SW, Lin CT, Lee SY, Huang HM. Slow freezing coupled static magnetic field exposure enhances cryopreservative efficiency--a study on human erythrocytes.. PLoS One 2013;8(3):e58988.
- Stoll C, Wolkers WF. Membrane Stability during Biopreservation of Blood Cells.. Transfus Med Hemother 2011;38(2):89-97.
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