超低温冷冻过程引起猪精子产生“似凋亡”变化

doi:10.11843/j.issn.0366-6964.2013.11.010

Abstract

Abstract:

The objective of the present work was to analyze whether cryopreservation could induce the “apoptosis-like” alteration of frozen-thawed boar sperm, and study the molecular mechanism of the fertility reduction of frozen-thawed sperm. Annexin-V/PI combined with the JC-1 fluorescence antibody Kit with flow cytometry (FCM) was used to determine the changes of phosphatidylserine (PS) location and mitochondrial membrane potential (Δψm) of boar sperm before and after cryopreservation with different incubation times . Content changes of “apoptosis” factors（Bcl-2，Caspase-9 and Caspase-3）in boar sperm were validated by immunoblotting analysis. The results showed that the motility of the cryopreserved sperm (54.4%) was lower than fresh sperm (87.9%) (P<0.05), in which the percentage of “apoptosis-like” of sperm with PS dislocation (26.8%) was 21.2% higher(P<0.05) than fresh sperm (5.6%). The cryopreservation caused an significant increase in the percentage of low Δψm of equilibrated and cryopreserved sperm (P<0.05).Compared with fresh sperm (19.6%), the percentage of sperm with low Δψm increased 59.8% after cryopreservation (P<0.05). The activating levels of apoptotic factors（Bcl-2，Caspase-9 and Caspase-3）in cryopreserved sperm were remarkably higher than the fresh and equilibrated sperm (P<0.05). Present evidences indicated that cryopreservation could induce spermatozoa into the “apoptosis-like” alteration. This study indicated that cryopreservation could not only lead to the dislocation of PS and reduced the mitochondrial membrane potential, but also stimulate the activities of apoptotic factors（Bcl-2,Caspase-9 and Caspase-3）, induce the frozen-thawed sperm into “apoptosis-like” state in advance, and shorten the survival time of frozen-thawed spermatozoa obviously. This might be another important reason for low pregnancy rate of frozen-thawed sperm.

CLC Number:

S828
S814.8

ZHAO Na, ZHEN Lin-qing, HU Qi-meng, WANG Liang-liang, LI Xin-hong. Cryopreservation Procedure Induces the Alterations of “Apoptosis-Like” of Boar Sperm[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(11): 1766-1774.

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Cryopreservation Procedure Induces the Alterations of “Apoptosis-Like” of Boar Sperm

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