ProAKAP4作为精子质量的蛋白质标记：来自冻融射精和附睾精子的见解

IF 2.5 2区农林科学 Q3 REPRODUCTIVE BIOLOGY

Theriogenology Pub Date : 2025-10-09 DOI:10.1016/j.theriogenology.2025.117698

Myrian Megumy Tsunokawa Hidalgo , Ana Beatriz Marques de Almeida , Leticia Amanda dos Santos Silva , Vinicius Wagner Silva , Julia Rodrigues Greghi , Natália Ribeiro Sambatti , Luiz Guilherme Corsi Trautwein , Rodrigo Yudi Palhaci Marubayashi , João Carlos Pinheiro Ferreira , Maria Isabel Mello Martins

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A total of fifteen adult mixed breed male cats between 1 and 3 years of age, with a mean weight of 4.21 ± 0.55 kg were included in this study. The first step of the study involved semen collection by urethral catheterization, using a sterile probe. In the second step, sperm were recovered from the epididymal tail after elective orchiectomy. The samples were cryopreserved in a TRIS-yolk extender. Thawed sperm straws were analyzed regarding their sperm kinematic, morphology and plasma membrane integrity. Sperm proAKAP4 concentration was determined using a commercial proAKAP4 ELISA kit. The relationship between proAKAP4 concentrations and sperm kinematics were evaluated by linear regressions, considering the proAKAP4 concentrations as independent variable and kinematic parameters as dependent variable. To compare concentration of proAKAP4 protein in both collection methods, paired <em>t</em>-test was used (P < 0.05). 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引用次数: 0

摘要

AKAP4蛋白是精子纤维鞘中含量最多的蛋白。该蛋白被认为有助于调节精子鞭毛运动，其前体proAKAP4被认为是不同物种精子质量的生物标志物。因此，本研究旨在评价proAKAP4浓度与猫精子的运动、形态以及冷冻保存的射精和附睾精子的膜完整性之间的关系。研究对象为15只1 ~ 3岁的成年杂交公猫，平均体重4.21±0.55 kg。研究的第一步是使用无菌探针通过尿道导尿收集精液。第二步，择期睾丸切除术后从附睾尾部提取精子。样品在tris -卵黄扩展器中冷冻保存。对解冻后的精子进行了运动学、形态学和质膜完整性分析。采用商用proAKAP4酶联免疫吸附测定试剂盒测定精子proAKAP4浓度。以proAKAP4浓度为自变量，运动学参数为因变量，采用线性回归方法评价proAKAP4浓度与精子运动学的关系。比较两种采集方法中proAKAP4蛋白的浓度，采用配对t检验（P < 0.05）。结果显示，精子中proAKAP4含量平均值（31.43±3.36 ng/mL, 15.26 ~ 59.18 ng/mL）高于附睾精子（19.58±1.67 ng/mL, 12.15 ~ 32.14 ng/mL）（P = 0.004）。可以观察到proAKAP4浓度与射精中TM （r2 = 0.574, P = 0.001）和dance （r2 = 0.267, P = 0.049）参数的关系。此外，proAKAP4浓度与附睾精子中TM （r2 = 0.670, P < 0.001）和静态精子细胞百分比（r2 = 0.477, P = 0.004）呈正相关。总之，我们的研究结果表明，proAKAP4浓度与解冻后射精和附睾精子的运动学参数相关，表明其在维持鞭毛结构和调节能力相关运动方面的作用。

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ProAKAP4 as a protein marker of sperm quality: Insights from frozen-thawed ejaculated and epididymal sperm

The AKAP4 protein is the most abundant protein of the sperm fibrous sheath. This protein is considered instrumental in regulating sperm flagellum motion and its precursor, proAKAP4, is considered a biomarker for sperm quality in different species. Therefore, this study aimed to evaluate the relationship of proAKAP4 concentration on feline sperm kinematic, morphology, and membrane integrity of cryopreserved ejaculated and epididymal sperm. A total of fifteen adult mixed breed male cats between 1 and 3 years of age, with a mean weight of 4.21 ± 0.55 kg were included in this study. The first step of the study involved semen collection by urethral catheterization, using a sterile probe. In the second step, sperm were recovered from the epididymal tail after elective orchiectomy. The samples were cryopreserved in a TRIS-yolk extender. Thawed sperm straws were analyzed regarding their sperm kinematic, morphology and plasma membrane integrity. Sperm proAKAP4 concentration was determined using a commercial proAKAP4 ELISA kit. The relationship between proAKAP4 concentrations and sperm kinematics were evaluated by linear regressions, considering the proAKAP4 concentrations as independent variable and kinematic parameters as dependent variable. To compare concentration of proAKAP4 protein in both collection methods, paired t-test was used (P < 0.05). When comparing proAKAP4 concentrations in ejaculated and epididymal sperm, it was observed that the mean of proAKAP4 in ejaculated sperm were higher, 31.43 ± 3.36 ng/mL (15.26–59.18 ng/mL) than epididymal sperm 19.58 ± 1.67 ng/mL (12.15–32.14 ng/mL) (P = 0.004). It was possible to observe the relationship between proAKAP4 concentration and the parameters: TM (r2 = 0.574, P = 0.001) and dance (r2 = 0.267, P = 0.049) in ejaculated sperm. Also, a relationship between the proAKAP4 concentration and TM (r2 = 0.670, P < 0.001) and the percentage of static sperm cells (r2 = 0.477, P = 0.004) in epididymal sperm was seen. In conclusion, our findings indicated that proAKAP4 concentrations are correlated with kinematic parameters in both ejaculated and epididymal sperm post-thaw, suggesting its role in sustaining flagellar architecture and modulating capacitation-associated motility of felines.

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来源期刊

Theriogenology 农林科学-生殖生物学

CiteScore

5.50

自引率

14.30%

发文量

387

审稿时长

72 days

期刊介绍： Theriogenology provides an international forum for researchers, clinicians, and industry professionals in animal reproductive biology. This acclaimed journal publishes articles on a wide range of topics in reproductive and developmental biology, of domestic mammal, avian, and aquatic species as well as wild species which are the object of veterinary care in research or conservation programs.