Vandenberghe L T M, Heindryckx B, Smits K, Szymanska K, Ortiz-Escribano N, Ferrer-Buitrago M, Pavani K, Peelman L, Deforce D, De Sutter P, Van Soom A, De Schauwer C
Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
Ghent-Fertility and Stem Cell Team (G-FaST), Department for Reproductive Medicine, Ghent University Hospital, 9000 Ghent, Belgium.
Reprod Fertil Dev. 2018 Nov;30(12):1739-1750. doi: 10.1071/RD18019.
Platelet-activating factor (PAF) is a well-described autocrine growth factor involved in several reproductive processes and is tightly regulated by its hydrolysing enzyme, PAF acetylhydrolase 1B (PAFAH1B). This intracellular enzyme consists of three subunits: one regulatory, 1B1, and two catalytic, 1B2 and 1B3. PAFAH1B3 has remained uncharacterised until now. Here, we report that PAFAH1B3 is present during the different stages of the first meiotic division in bovine, murine and human oocytes. In these species, the PAFAH1B3 subunit was clearly present in the germinal vesicle, while at metaphase I and II, it localised primarily at the meiotic spindle structure. In cattle, manipulation of the microtubules of the spindle by nocodazole, taxol or cryopreservation revealed a close association with PAFAH1B3. On the other hand, disruption of the enzyme activity either by P11, a selective inhibitor of PAFAH1B3, or by PAFAH1B3 antibody microinjection, caused arrest at the MI stage with defective spindle morphology and consequent failure of first polar body extrusion. In conclusion, our results show that one of the catalytic subunits of PAFAH1B, namely PAFAH1B3, is present in bovine, murine and human oocytes and that it plays a functional role in spindle formation and meiotic progression during bovine oocyte maturation.
血小板活化因子(PAF)是一种已被充分描述的自分泌生长因子,参与多种生殖过程,并受到其水解酶——血小板活化因子乙酰水解酶1B(PAFAH1B)的严格调控。这种细胞内酶由三个亚基组成:一个调节亚基1B1和两个催化亚基1B2和1B3。直到现在,PAFAH1B3仍未被充分了解。在此,我们报告PAFAH1B3存在于牛、鼠和人类卵母细胞第一次减数分裂的不同阶段。在这些物种中,PAFAH1B3亚基在生发泡中清晰可见,而在减数分裂中期I和II时,它主要定位于减数分裂纺锤体结构上。在牛中,用诺考达唑、紫杉醇或冷冻保存处理纺锤体微管,发现其与PAFAH1B3密切相关。另一方面,通过PAFAH1B3的选择性抑制剂P11或显微注射PAFAH1B3抗体破坏酶活性,会导致减数分裂中期I停滞,纺锤体形态异常,进而导致第一极体排出失败。总之,我们 的结果表明,PAFAH1B的一个催化亚基,即PAFAH1B3,存在于牛、鼠和人类卵母细胞中,并且在牛卵母细胞成熟过程中的纺锤体形成和减数分裂进程中发挥功能作用。