Effect of cryopreservation on the expression of sperm surface protein PH-20 and ratio of apoptosis in normal birth and infertility

doi:10.16098/j.issn.0529-1356.2017.01.015

Abstract

Abstract:

Objective To investigate the effect of cryopreservation on the expression of sperm surface protein PH-20 and the sperm apoptosis. Methods Semen samples were obtained from fertile men (n=14, group A) and infertile men (n=20, group B). Western blotting was used to detect the PH-20 protein expression in human spermatozoa.The localization of this protein on human spermatozoa was determined by indirect immunofluorescent staining using PH-20 antibody. The sperm apoptosis was examined by terminal deoxynucleotidyl transferase(TdT) mediated deoxyuridine triphophate-biotin nick end labeling(TUNEL). Results After cryopreservation, the level of PH-20 protein expression was significantly lower in both group A and B than that of fresh sperm (P<0.05). The percentage of PH-20 positive rate was significantly lower in both group A and B than that of fresh sperm (P<0.05). The ratio of sperm apoptosis was not significantly different in thawed group A than that of fresh sperm (P>0.05). The ratio of sperm apoptosis was significantly lower in thawed group B than that of fresh sperm (P<0.05).
Conclusion Cryopreservation caused significant reduction of PH-20 protein expression, the percentage of PH-20 positive rate in human sperm. But the cryopreservation had no significant influence on normal fertility of sperm apoptosis.

Key words: Sperm, Cryopreservation, PH-20, Western blotting, Human

MA Xiao-ping GAO Xiao-qin ZHOU Hua WANG Jun. Effect of cryopreservation on the expression of sperm surface protein PH-20 and ratio of apoptosis in normal birth and infertility[J]. Acta Anatomica Sinica, 2017, 48(1): 87-91.

References

［1］Taggart DA, Leigh CM, Steele VR, et al. Effect of cooling and cryopreservation on sperm motility and morphology of several species of marsupial ［J］. Reprod Fertil Dev, 1996, 8(4):673-679.

［2］Belker AM, Bergamini DA. The feasibility of cryopreservation of sperm harvested intraoperatively during vasectomy reversals ［J］. J Urol, 1997, 157(4):1292-1294.

［3］Morris GJ, Acton E, Avery S. A novel approach to sperm cryopreservation ［J］. Hum Reprod, 1999, 14(4):1013-1021.

［4］Wassarman PM. Mammalian fertilization: egg and sperm (glyco) proteins that support gamete adhesion［J］. Am J Reprod Immunol, 1995, 33(3):253-258.

［5］Zilli L, Schiavone R, Zonno V, et al. Effect of cryopreservation on sea bass sperm proteins［J］. Biol Reprod, 2009, 72(5):1262-1267.

［6］WHO. Laboratory Manual for the Examination of Human Semen and Sperm- Cervical Mucus Interaction［M］. 4th ed. London: Cambridge University Press,1999: 112-113.

［7］Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding［J］. Anal Biochem, 1976, 72:248-254.［8］Critser JK，Mobraaten LE. Cryopreservation of murine spermatozoa［J］. ILAR J, 2000,41(4):197-206.

［9］Desrosiers P, Légaré C, Leclerc P, et al. Membranous and structural damage that occur during cryopreservation of human sperm may be time-related events［J］. Fertil Steril, 2006, 85(6):1744-1752.

［10］Carl L, Sophie P, Pierre L, et al. Cryopreservation alters the levels of the bull sperm surface protein P25b［J］. J Androl, 2000, 21(5):700-707.

［11］Cherr GN, Yudin AI, Overstreet JW. The dual function of GPI-anchored PH-20: hyaluronidase and intracellular signaling［J］. Matrix Biol, 2001, 20(8): 515-525.

［12］Deng X, He Y, Martin-Deleon PA. Mouse spam1(PH-20): evidencefor its expression in the epididymis and for a new category of spermatogenic expressed gene［ J ］. J Androl, 2000, 21(6):822-832.

［13］Rutllant J, Meyers SA. Posttranslational processing of PH-20 during epididymal sperm maturation in the horse ［J］. Biol Reprod, 2001, 65(5): 1324-1331.

［14］Wang WM, Dong YF, Zhang ChJ. The progress of research on human sperm DNA damage and repair ［J］. Chinese Journal of Healthy Birth and Heredity, 2015, (5) : 12-14.(in Chinese)

王伟明, 董毅飞, 张昌军. 人类精子DNA损伤与修复的研究进展［J］. 中国优生与遗传杂志, 2015, (5):12-14.

［15］Huang DH, Xiong ChL. Effects of cryopreservation on human sperm genetic substance ［J］. National Journal of Andrology, 2002, 8(2): 218-220. (in Chinese)

黄东晖, 熊承良. 冷冻对人精子遗传物质的影响［J］. 中华男科学杂志, 2002, 8(2):218-220.

［16］Lin QL. The influence of cryopreservation on human sperm nuclear DNA integrity and the study of improved human sperm quality using antioxidants supplementation in semen cryopreservation procedure ［D］. Shantou University, 2007. (in Chinese)

林琼林. 冷冻对人精子核DNA完整性的影响及抗氧化剂改善冻融人精子质量的研究［D］. 汕头大学, 2007.

［17］Kwon IK, Park KE, Niwa K.Activation, pronuclear formation, and development in vitro of pig oocytes following intracytoplasmic injection of freeze-dried spermatozoa［J］. Biol Reprod, 2004, 71(5):951-958.

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