Inhibition of PDE3A sustains meiotic arrest and gap junction of bovine growing oocytes in in vitro growth culture
Inhibition of PDE3A sustains meiotic arrest and gap junction of bovine growing oocytes in in vitro growth culture
复制标题
DOI：
10.1016/j.theriogenology.2018.05.028
复制
发表时间：
2018-09-15
期刊：
影响因子：
2.8
通讯作者：
Miyano, Takashi
中科院分区：
文献类型：
--
作者：
Alam, Md Hasanur;Lee, Jibak;Miyano, Takashi
牛早期有腔卵泡（1.2- 1.8mm）中直径为105-115 μ m的卵母细胞能够恢复减数分裂，但缺乏成熟到中期II的能力。为了赋予卵母细胞完全成熟能力，需要能够支持其生长并防止其在培养期间减数分裂恢复的培养系统。在这项研究中，我们培养生长的卵母细胞5天，研究不同的磷酸二酯酶（PDE）抑制剂对减数分裂阻滞和收购的完全成熟能力的生长的卵母细胞，和他们的间隙连接通信与卵丘细胞的影响。用3-异丁基-1-甲基黄嘌呤（IBMX;广谱PDE抑制剂）、咯利普兰（PDE 4抑制剂）、西洛酰胺和米力农（PDE 3A抑制剂）培养生长的卵母细胞-卵丘复合体（OCC）。各组卵母细胞平均直径增加相似。IBMX，西洛酰胺和米力农诱导胃窦形成OCC和维持减数分裂停滞的卵母细胞在培养过程中，而咯利普兰既不促进胃窦形成，也不维持卵母细胞减数分裂停滞。卵母细胞和卵丘细胞之间的缝隙连接通信由IBMX和西洛酰胺维持，但不是由咯利普兰作为判断从卵母细胞到卵丘细胞的注射荧光黄染料的转移。在随后的体外成熟，卵母细胞生长与IBMX，西洛酰胺和米力农表现出完全的成熟能力。这些结果表明，PDE 3A抑制维持牛生长的卵母细胞的减数分裂停滞，并维持其与卵丘细胞的间隙连接通讯5天，从而有助于其获得完全成熟能力。(C)2018爱思唯尔公司All rights reserved.
Bovine growing oocytes with a diameter of 105-115 mu m from early antral follicles (1.2-1.8 mm) are able to resume meiosis, but lack the competence to mature to metaphase II. To confer full maturation competence onto the oocytes, culture systems which can support their growth and prevent their meiotic resumption during culture are needed. In this study, we cultured growing oocytes for 5 days to examine the effects of different phosphodiesterase (PDE) inhibitors on meiotic arrest and acquisition of full maturation competence of growing oocytes, and their gap junctional communication with cumulus cells. Growing oocyte-cumulus complexes (OCCs) were cultured with 3-isobutyl-1-methylxanthine (IBMX; broad-spectrum PDE inhibitor), rolipram (PDE4 inhibitor), cilostamide and milrinone (PDE3A inhibitors). The mean diameters of oocytes increased similarly in all groups. IBMX, cilostamide and milrinone induced antrum formation by OCCs and maintained meiotic arrest of oocytes during culture, whereas rolipram neither promoted antrum formation nor maintained oocyte meiotic arrest. Gap junctional communication between oocytes and cumulus cells was maintained by IBMX and cilostamide, but not by rolipram as judged by the transfer of injected lucifer yellow dye from oocytes to cumulus cells. In subsequent in vitro maturation, oocytes grown with IBMX, cilostamide and milrinone showed full maturation competence. These results suggest that PDE3A inhibition maintains the meiotic arrest of bovine growing oocytes and sustains their gap junctional communication with cumulus cells for 5 days, thereby contributing to their acquisition of full maturation competence. (C) 2018 Elsevier Inc. All rights reserved.