Dataset Information

Mechanisms of ROS-induced mitochondria-dependent apoptosis underlying liquid storage of goat spermatozoa

ABSTRACT: Liquid storage of spermatozoa is important for artificial insemination and herd genetic breeding. However, the extending time of storage inducing rapid decline of spermatozoa quality limits the development of this technology. The molecular mechanisms underlying liquid storage of spermatozoa remain largely unexplored. In this study, the effects of liquid storage on functional quality of spermatozoa were assessed in goat (Capra hircus). The time-dependent decline of spermatozoa motility showed a strong correlation with the significant increase of apoptosis. Moreover, apoptosis-related ultrastructural changes were observed, especially the defect in mitochondria. Meanwhile, significant decrease of mitochondrial membrane potential and expression changes of mitochondrial apoptosis-related proteins indicated mitochondrial dysfunction and activation of mitochondrial apoptotic pathway. Notably, the abnormal high level of reactive oxygen species (ROS) caused by liquid storage resulted in the oxidative damage to mitochondria and accelerated mitochondria-dependent apoptosis, which was demonstrated by addition of ROS scavenger N-acetylcysteine. Furthermore, critical differentially expressed proteins involved in mitochondria-dependent apoptosis and antioxidant defense were identified and profiled by quantitative proteomic analysis, facilitating the understanding of molecular regulation of ROS-induced mitochondria-dependent apoptosis. These outcomes provide insights into the mechanisms underlying liquid storage of goat spermatozoa and enhance the progress of semen storage technology.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Capra Hircus (goat)

TISSUE(S): Spermatozoon

SUBMITTER: Tengfei Liu

LAB HEAD: Tengfei Liu

PROVIDER: PXD014609 | Pride | 2019-11-12

REPOSITORIES: Pride

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Mechanisms of ROS-induced mitochondria-dependent apoptosis underlying liquid storage of goat spermatozoa.

Liu Tengfei T Han Yawen Y Zhou Ting T Zhang Ruihang R Chen Hong H Chen Shulin S Zhao Huiying H

Aging 20190923 18

Liquid storage of spermatozoa is important for artificial insemination and herd genetic breeding. However, the extended time of storage inducing the rapid decline in spermatozoa quality limits the development of this technology. The molecular mechanisms underlying liquid storage of spermatozoa remain largely unexplored. In this study, the effects of liquid storage on functional quality of spermatozoa were assessed in goat (<i>Capra hircus</i>). The time-dependent decline in spermatozoa motility ...[more]

PMID: 31548434

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Project description:Background: The cryopreservation of semen is crucial for conserving genetic diversity and reproductive success in endangered species like the Siberian sturgeon. However, this process can cause cryodamage, affecting quality and protein profile of spermatozoa. While cryoprotectants like dimethyl sulfoxide (DMSO) and methanol (MeOH) facilitate post-thaw motility recovery, DMSO-preserved spermatozoa exhibit reduced fertilizing ability. This study investigates how DMSO and MeOH impact the proteome of Siberian sturgeon spermatozoa and examines semen quality parameters. Two complementary approaches of quantitative proteomics, liquid chromatography-mass spectrometry (LC-MS) and two-dimensional difference in gel electrophoresis (2D-DIGE), were used to analyze the proteomic profiles of fresh and cryopreserved spermatozoa, as well as the extracellular medium (EM; n=7 for each group). Results: Cryopreservation led to a decline in motility parameters (MOT, VCL, PROG) and viability, along with an increase in ROS levels, membrane fluidity, and acrosome damage. Despite similar quality parameters between DMSO and MeOH-preserved sperm, DMSO-preserved sperm showed dramatically lower fertilization success (6.2% vs 51.2%). A total of 224 and 118 differentially abundant proteins in spermatozoa cryopreserved with MeOH and DMSO, respectively, were identified compared to fresh samples, with 342 and 363 proteins released into the EM. The most affected proteins by cryopreservation included H2A, CABYR, PYGM, ENO3, DBI and LTA4H. Additionally, 36 and 39 uniquely altered sperm-leakage proteins were identified for MeOH and DMSO cryopreserved samples, respectively. Bioinformatic analysis showed that MeOH-specific proteins were related to chromosomal structure and mitochondrial functionality, while DMSO-specific proteins were mainly involved in acrosome reaction, zona pellucida binding, flagella structure, and nuclear pore organization. These proteins are potentially involved in sturgeon sperm fertilizing ability. The expression of six proteins was verified by western blot analysis. Conclusions: This study provides the first comprehensive proteomic characterization of Siberian sturgeon spermatozoa after cryopreservation with DMSO and MeOH, revealing insights into proteomic changes that affect fertilizing ability and aiding conservation efforts for this endangered species.

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