Effects of cryoprotectant treatments on bovine sperm function and osmolyte content  

冷冻保护剂对牛精子功能和渗透压的影响

作　　者：Erif E. M. Setyawan Trevor G. Cooper Dyah A. Widiasih Aris Junaidi Ching-Hei Yeung 

机构地区：[1]Centre of Reproductive Medicine and Andrology of the University Clinic, Miinster D-48149, Germany [2]Department of Obstetrics and Gynecology, Faculty of Veterinary Medicine, Gadjah Mada University, Yogyakarta 55281,Indonesia [3]Department of Public Health, Faculty of Veterinary Medicine, Gadjah Mada University, Yogyakarta 55281, Indonesia

出　　处：《Asian Journal of Andrology》2009年第5期571-582,I0002,I0003,共14页亚洲男性学杂志（英文版）

摘　　要：The hypothesis that addition and removal of cryoprotectants to and from spermatozoa would initiate regulatory volume decrease, and lead to osmolyte loss and reduced sperm function, was tested. Common cryoprotectants, in the absence of freezing and thawing, affected bovine ejaculated spermatozoa by lowering their total and progressive motility in medium, reducing their migration through surrogate cervical mucus, damaging sperm head membranes and inducing sperm tail coiling. Sperm function was slightly better maintained after cryoprotectants were added and removed in multiple small steps rather than in a single step. The intracellular content of the polyol osmolytes, D-sorbitol and myo-inositol, exceeded that of the zwitterion osmolytes, L-carnitine and L-glutamate. Certain cryoprotectants reduced intracellular L-camitine and L-glutamate concentration but not that of myo-inositol or D-sorbitol. Multistep treatments with some cryoprotectants had advantages over one-step treatments in mucus penetration depending on the original amount of intracellular camitine and glutamate in the spermatozoa. Overall, sperm quality was best maintained by multistep treatment with glycerol and propanediols that were associated with decreased intracellular glutamate concentration. Bovine spermatozoa seem to use glutamate to regulate cryoprotectant-induced cell swelling.The hypothesis that addition and removal of cryoprotectants to and from spermatozoa would initiate regulatory volume decrease, and lead to osmolyte loss and reduced sperm function, was tested. Common cryoprotectants, in the absence of freezing and thawing, affected bovine ejaculated spermatozoa by lowering their total and progressive motility in medium, reducing their migration through surrogate cervical mucus, damaging sperm head membranes and inducing sperm tail coiling. Sperm function was slightly better maintained after cryoprotectants were added and removed in multiple small steps rather than in a single step. The intracellular content of the polyol osmolytes, D-sorbitol and myo-inositol, exceeded that of the zwitterion osmolytes, L-carnitine and L-glutamate. Certain cryoprotectants reduced intracellular L-camitine and L-glutamate concentration but not that of myo-inositol or D-sorbitol. Multistep treatments with some cryoprotectants had advantages over one-step treatments in mucus penetration depending on the original amount of intracellular camitine and glutamate in the spermatozoa. Overall, sperm quality was best maintained by multistep treatment with glycerol and propanediols that were associated with decreased intracellular glutamate concentration. Bovine spermatozoa seem to use glutamate to regulate cryoprotectant-induced cell swelling.

关 键 词：cryoconservation CRYOPROTECTANTS osmotic swelling regulatory volume decrease 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]