Plasma Membrane Permeability of Carp (Cyprinus carpio, L., 1758) Spermatozoa for Water and Cryoprotectants Molecules at Different Stages of Cryopreservation
Keywords:carp spermatozoa, membrane permeability, activation energy, cryopreservation, ethylene glycol, glycerol, 1, 2-propanediol, dimethyl sulfoxide
Cell membrane permeability for water and cryoprotectant molecules is an important parameter to determine cooling rates during fish genetic material cryopreservation. Cell volume kinetics was assessed spectrofotometrically, the resulted experimental dependences of relative cell volume vs. time were fitted with solutions of theoretical model equations that allowed us to calculate the membrane permeability coefficients of carp spermatozoa for molecules of water and several cryoprotectants. The plasma membrane permeability coefficient of carp spermatozoa at 20°C was established to make (3.05 ± 0.40)×10<sup>–14</sup> m<sup>3</sup>/ N·s, but when it decreased within 35...15°C temperature range the activation energy equaled to (53.9 ± 3.8) kJ/mol. A decreased membrane permeability of carp spermatozoa for dimethyl sulfoxide, ethylene glycol and 1,2-propanediol molecules within the mentioned range was characterized by the activation energy of 70–80 kJ/mol. This fact indicated that molecules of the studied substances penetrated into spermatozoon via a passive diffusion through a lipid bilayer. Our findings could be used to determine the optimal cryopreservation regimen for carp spermatozoa.
Probl Cryobiol Cryomed 2016; 26(4): 340–348
Boryshpolets S., Dzyuba B., Rodina M. et al. Freeze-thawing as the factor of spontaneous activation of spermatozoa motility in common carp (Cyprinus carpio L.). Cryobiology 2009; 59: 291–296. CrossRef PubMed
Cabrita E., Alvarez R., Anel E., Herraez M.P. The hypoosmotic swelling test performed with coulter counter: a method to assay functional integrity of sperm membrane in rainbow trout. Anim Reprod Sci 1999; 55: 279–287. CrossRef
Chernobay N.A., Kovalenko I.F., Koschiy S.V. et al. Temperature dependence of SPEV cell membranes permeability for molecules of cryoprotectants. Problems of Cryobiology 2011; 21(1): 46–51.
Fauvel C., Suquet M., Cosson J. Evaluation of fish sperm quality. Appl Ichthyol 2010; 26: 636–643. CrossRef
Gordiyenko O.I., Kovalenko S.Ye., Kovalenko I.F. Mechanisms of glycerol permeability through the membrane of human erythrocytes. Probl Cryobiol 2012; 22(4): 389–397.
Gordiyenko Ye.A., Pushkar N.S. Physics basis of low temperature preservation of cell suspensions. Kyiv: Naukova dumka; 1994.
Gordiyenko Ye.A., Tovstyak V.V. Physics of biomembranes. Kyiv: Naukova dumka; 2009.
Kopeika E.F. Ecological niche as the factor determining cryoresistance in fish spermatozoa. Probl Cryobiol Cryomed 2014; 24(4): 302–311. CrossRef
Kopeika E.F. Manual on carp sperm low temperature preservation. Moscow: VNIIPRH; 1986.
Kopeika E.F., Kopeika J.E. Variability of sperm quality after cryopreservation in fish. In: Alavi S.H.M., Cosson J., Coward K., Rafiee G., editors. Fish spermatology. Oxford: Alpha Science International Ltd; 2007. p. 347–396.
Kopeika E., Kopeika J., Zhang T. Cryopreservation of fish sperm In: Day J.G. Stacey G.N., editors. Cryopreservation and freezedrying protocols. Totowa, New Jersey: Humana Press; 2007. p. 203–217.
Meryman H.T. Osmotic stress as a mechanism of freezing injury. Cryobiology 1971; 8(5): 489–500. CrossRef
Oldenhof H., Friedel K., Sieme H. et al. Membrane permeability parameters for freezing of stallion sperm as determined by Fourier transform infrared spectroscopy. Cryobiology 2010; 61: 115–122. CrossRef PubMed
Petrunkina A. M. Fundamental aspects of gamete cryobiology. J Reprod Med Endocrinol 2007; 4: 78–91.
Pinisetty D., Huang C., Dong Q. et al. Subzero water permeability parameters and optimal freezing rates for sperm cells of the southern platyfish, Xiphophorus maculates. Cryobiology 2005; 50: 250–263. CrossRef PubMed
Puhovkin A.Yu., Kononenko I.S., Cherepnin V.O. et al. Permeability of sterlet sperm membranes (Acipenser ruthenus L., 1758) for water molecules. Fisheries Science of Ukraine 2016; 35(1): 70–77.
Puhovkin A.Yu., Kopeika E.F., Gordiyenko E.A., Nardid O.A., inventors. The method for determining the permeability of membranes of common carp sperm to the water molecules. Patent of Ukraine № 104809 IPC G01N 33/48, G01N 15/00. 2014 March 11.
Puhovkin A.Yu., Kopeika E.F., Nardid O.A., Cherkashina Ya.O. Investigation of membrane permeability of carp spermatozoa for water molecules. Biophysics 2014; 59(3): 481–487.
Puhovkin A.Yu., Kopeika E.F. Study of water molecules transfer through pike (Esox lucius L) spermatozoa membranes. Probl Cryobiol Cryomed 2015; 25(2): 165. CrossRef
Rana K.J., Gilmour A. Cryopreservation of fish spermatozoa: effect of cooling methods on the reproducibility of cooling rates and viability. In: Refrigeration and Aquaculture Conference, Bordeaux, 20–22 March 1996.
Suquet M., Dreanno C., Fauvel C. et al. Cryopreservation for sperm in marine fish. In: Refrigeration and Aquaculture Conference, Bordeaux, 2000; 31: 231–243. CrossRef
Verma D.K., Routray P., Dash C. et al. Physical and biochemical characteristics of semen and ultrastructure of spermatozoa in six carp species. Turkish J Fish Aquat Sci 2009; 9: 67–76.
Zhang T.T. Cryopreservation of gametes and embryos of aquatic species. In: Fuller B.J., Lane N., Benson E.E., editors. Life in the Frozen State. Florida: CRC Press LLC; 2004. p. 415–436.
How to Cite
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).