Kashir Junaid, Mistry Bhavesh V, BuSaleh Lujain, Nomikos Michail, Almuqayyil Sarah, Abu-Dawud Raed, AlYacoub Nadya, Hamdan Hamdan, AlHassan Saad, Lai F Anthony, Assiri Abdullah M, Coskun Serdar
Department of Biology, College of Arts and Sciences, Khalifa University, Abu Dhabi 127788, United Arab Emirates.
Department of Comparative Medicine, King Faisal Specialist Hospital and Research Centre, Riyadh 11564, Saudi Arabia.
Pharmaceuticals (Basel). 2023 Jan 28;16(2):198. doi: 10.3390/ph16020198.
Mammalian oocyte activation is initiated by intracellular calcium (Ca) oscillations, driven by the testis-specific phospholipase C zeta (PLCζ). Sperm PLCζ analysis represents a diagnostic measure of sperm fertilisation capacity. The application of antigen unmasking/retrieval (AUM) generally enhanced the visualisation efficacy of PLCζ in mammalian sperm, but differentially affected the PLCζ profiles in sperm from different human males. It is unclear whether AUM affects the diagnosis of PLCζ in human sperm. Herein, we examined whether the application of AUM affected the correlation of PLCζ profiles with sperm parameters and fertilisation capacity. PLCζ fluorescence levels and localisation patterns were examined within the sperm of males undergoing fertility treatment (55 patients aged 29-53) using immunofluorescence in the absence/presence of AUM. The changes in PLCζ profiles following AUM were examined in relation to sperm health and fertilisation outcome. AUM enhanced the observable levels and specific localisation patterns of PLCζ in relation to both optimal sperm parameters and fertilisation outcome, without which significant differences were not observed. The extent of the change in levels and localisation ratios of PLCζ was also affected to a larger degree in terms of the optimal parameters of sperm fertility and fertilisation capacity by AUM. Collectively, AUM was essential to accurately assesses PLCζ in human sperm in both scientific and clinical contexts.
哺乳动物卵母细胞激活由睾丸特异性磷脂酶Cζ（PLCζ）驱动的细胞内钙（Ca）振荡启动。精子PLCζ分析是精子受精能力的一种诊断手段。抗原暴露/修复（AUM）的应用通常会增强PLCζ在哺乳动物精子中的可视化效果，但对不同男性精子中的PLCζ图谱有不同影响。目前尚不清楚AUM是否会影响人类精子中PLCζ的诊断。在此，我们研究了AUM的应用是否会影响PLCζ图谱与精子参数和受精能力之间的相关性。在有/无AUM的情况下，使用免疫荧光法检测接受生育治疗的男性（55例，年龄29 - 53岁）精子内的PLCζ荧光水平和定位模式。研究了AUM后PLCζ图谱的变化与精子健康和受精结果的关系。AUM增强了与最佳精子参数和受精结果相关的PLCζ的可观察水平和特定定位模式，否则未观察到显著差异。AUM在精子生育力和受精能力的最佳参数方面，对PLCζ水平和定位比率变化程度的影响也更大。总的来说，AUM对于在科学和临床背景下准确评估人类精子中的PLCζ至关重要。
