猪冷冻精子的细胞凋亡及凋亡途径初步研究

doi:10.11843/j.issn.0366-6964.2015.08.009

Abstract

Abstract:

In order to investigate the effects of cryopreservation on boar spermatozoa apoptosis and apoptotic pathway，the flow cytometry was used to detect the apoptosis state and ROS level of boar frozen spermatozoa.ATP content was measured by luminometer，and the mRNA expression level of genes involved in different apoptotic pathways were measured by real-time quantitative RT-PCR method.The results showed that the motility，normal acrosome rate and plasma membrane integrity rate of sperm after freezing were decreased greatly (P<0.05).The TUNEL-positive spermatozoa rate (80.4%) from freezing group were much higher than that of fresh group (9.7%) (P<0.05).The percentage of ROS-positive spermatozoa in freezing group (58.2%) was also obviously higher than that of fresh group (P<0.05).The cryopreservation caused a significant decrease of ATP concentration in freezing group (0.247 μmol•L^-1) comparing with fresh group (0.926 μmol•L^-1) (P<0.05).qRT-PCR results showed that the relative expression level from promoting apoptosis genes (TNF-α，Fas，Caspase，P53 and Bax) were up-regulated，and in which the expression of Fas，P53，TNF-α，Caspase-8 and Caspase-9 increased greatly after cryopreservation (P<0.05).The relative expression level from inhibiting apoptosis genes (BCL-2，BIRC-5，MnSOD，CuZnSOD and SURVIVN) were down-regulated，and in which the expression of BIRC-5，MnSOD and SURVIVN decreased greatly (P<0.05).The results indicated that cryopreservation could lead to boar spermatozoa apoptosis.Not only extrinsic death receptor but also intrinsic mitochondrial signaling pathway might play key roles in mediating spermatozoa apoptosis after cryopreservation.

CLC Number:

S828.2

WANG Xin，DAI Jian-jun，WU Cai-feng,ZHANG Shu-shan，WU Yun-long，ZHANG De-fu. Preliminary Studies on Apoptosis and Apoptotic Pathways of Frozen Boar Spermatozoa[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(8): 1341-1347.

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Preliminary Studies on Apoptosis and Apoptotic Pathways of Frozen Boar Spermatozoa

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