Optimizing thawing conditions are required to maximize recovery of semen capacities after cryopreservation. This study aimed to assess the effect of thawing procedures on motility recovery and maintenance of semen capacities over time. A fractional factorial design approach was used to reduce the number of repetitions and simultaneously analyze the different interactions. Thirty canine frozen semen samples were thawed under different thawing conditions. Motility and velocity parameters were recorded using a computer-assisted semen analyzer up to 6 hours after thawing. Ten quadratic models were found to be significant, with the most significant effects observed on the velocity parameters. Second, this study allowed us to evaluate the effect of the proAKAP4 marker on frozen/thawed semen, with regard to motility recovery. ProAKAP4 is the precursor of A-kinase anchor protein 4. It has been studied in several species as a marker to assess sperm quality. In our study, the expression of proAKAP4 was determined using flow cytometry. No correlation was found between post-thaw motility parameters and proAKAP4 levels. However, the effects of thawing temperature, incubation time, and straw size were significant and similar to those observed for velocity parameters.

中文翻译：

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使用新的 A-激酶锚定蛋白 4 前体生物标志物评估优化的犬类冷冻/解冻精液样本。

需要优化解冻条件以最大限度地恢复冷冻保存后的精液容量。本研究旨在评估解冻程序随时间推移对精子活力恢复和维持能力的影响。部分因子设计方法用于减少重复次数并同时分析不同的相互作用。三十个犬冷冻精液样本在不同解冻条件下解冻。使用计算机辅助精液分析仪在解冻后 6 小时内记录运动和速度参数。发现十个二次模型是显着的，对速度参数的影响最为显着。其次，这项研究使我们能够评估 proAKAP4 标记对冷冻/解冻精液的影响，关于运动恢复。ProAKAP4 是 A 激酶锚定蛋白 4 的前体。它已在多个物种中作为评估精子质量的标记物进行了研究。在我们的研究中，使用流式细胞术测定 proAKAP4 的表达。在解冻后运动参数和 proAKAP4 水平之间没有发现相关性。然而，解冻温度、孵化时间和秸秆大小的影响是显着的，与观察到的速度参数相似。