Identification and Validation of Yak (Bos grunniens) Frozen–Thawed Sperm Proteins Associated with Capacitation and the Acrosome Reaction

To
achieve fertilization, mammalian spermatozoa must undergo capacitation
and the acrosome reaction (AR) within the female reproductive tract.
However, the effects of cryopreservation on sperm maturation and fertilizing
potential have yet to be established. To gain insight into changes
in protein levels within sperm cells prepared for use in the context
of fertilization, a comprehensive quantitative proteomic profiling
approach was used to analyze frozen–thawed Ashidan yak spermatozoa
under three sequential conditions: density gradient centrifugation-based
purification, incubation in a capacitation medium, and treatment with
the calcium ionophore A23187 to facilitate AR induction. In total,
3280 proteins were detected in these yak sperm samples, of which 3074
were quantified, with 68 and 32 being significantly altered following
sperm capacitation and AR induction. Differentially abundant capacitation-related
proteins were enriched in the metabolism and PPAR signaling pathways,
while differentially abundant AR-related proteins were enriched in
the AMPK signaling pathway. These data confirmed a role for superoxide
dismutase 1 (SOD1) as a regulator of sperm capacitation while also
offering indirect evidence that heat shock protein 90 alpha (HSP90AA1)
regulates the AR. Together, these findings offer a means whereby sperm
fertility-related marker proteins can be effectively identified. Data
are available via Proteome Xchange with identifier PXD035038.

为了实现受精，哺乳动物的精子必须在女性生殖道中经历成熟化以及发生顶体反应（AR）。然而，冷冻保存对精子成熟及受精潜能的影响尚未得到明确。为了深入探究用于受精目的的精子细胞内蛋白质水平的变化，本研究采用了一种全面的定量蛋白质组学分析策略，以分析在三个连续条件下冷冻-解冻的阿什坦牦牛精子：基于密度梯度离心纯化、在成熟化培养基中孵育，以及使用钙离子载体A23187处理以促进AR诱导。总计检测到3280种蛋白质，其中3074种得到量化，68种和32种在精子成熟化和AR诱导后发生显著变化。与成熟化相关的差异丰度蛋白质富集于代谢和PPAR信号通路，而与AR相关的差异丰度蛋白质富集于AMPK信号通路。这些数据证实了超氧化物歧化酶1（SOD1）在精子成熟化过程中的调控作用，同时也为热休克蛋白90α（HSP90AA1）调控AR提供了间接证据。综合这些发现，为有效识别与精子生育能力相关的标记蛋白质提供了一种途径。数据可通过Proteome Xchange获取，标识符为PXD035038。