The potential role of gap junction communication between cumulus cells and bovine oocytes during in vitro maturation
The potential role of gap junction communication between cumulus cells and bovine oocytes during in vitro maturation
复制标题
DOI：
10.1002/mrd.20281
复制
发表时间：
2005-07-01
影响因子：
2.5
通讯作者：
Marc-André, S
中科院分区：
文献类型：
--
作者：
Atef, A;François, P;Marc-André, S
我们实验室的初步研究表明，在培养过程中调节卵丘的早期或晚期的扩张对卵丘包裹的卵母细胞的后续发育有着深远的影响。本研究旨在探讨牛卵母细胞体外成熟过程中短期暴露重组人卵泡刺激素(r-hFSH)对卵丘扩张和卵母细胞发育能力的影响。体外卵母细胞体外成熟后的囊胚发育率在培养的前6小时内有极显著提高(P<；0.0001)。为了证明卵母细胞和卵丘之间的功能性偶联在6小时内的重要性，卵丘-卵母细胞复合体(COCs)在r-hFSH的作用下成熟6小时，然后在1-庚醇或1-辛醇(缝隙连接抑制剂)的存在下成熟18小时，以阻断两者之间的通讯。加入偶联抑制剂后，囊胚率显著降低(P<；0.05)。用弱碱性甲胺短暂处理(30分钟)，可显著逆转缝隙连接抑制剂的作用(P<；0.05)，显著逆转这些药物对囊胚率的抑制作用。通过显微注射荧光染料荧光黄，进一步研究了卵母细胞与周围卵丘细胞之间的缝隙连接通讯。获得的形态证据(染料转移)支持在FSH短时间暴露下较长时间内存在功能偶联。总之，短期暴露于r-hFSH可获得较高的牛卵母细胞发育率。这种影响被认为是通过缝隙连接介导的，因为卵母细胞的发育能力因其功能的抑制而受到损害。(C)2005年Wiley-Liss，Inc.
Preliminary studies in our laboratory have indicated that modulating cumulus expansion early or late during culture has a profound influence on the subsequent development of cumulus-enclosed oocytes. Our objectives were to evaluate the effect of short term exposure to recombinant human follicle-stimulating hormone (r-hFSH) during in vitro maturation (IVM) on cumulus expansion and developmental competence of bovine oocytes. A highly significant (P < 0.0001) improvement in blastocyst development rate as a proportion of cleaved oocytes after IVM of oocytes was observed in the presence of r-hFSH for the first 6 hr of culture. To demonstrate the importance of the functional coupling between the oocyte and the cumulus compartment during that period of 6 hr, cumulus-oocyte complexes (COCs) were matured with r-hFSH for the first 6 hr followed by 18 hr in presence of 1-heptanol or 1-octanol (gap junction inhibitors) to block the communication between the two. With the coupling inhibitors, the blastocyst yield was significantly decreased (P < 0.05). A brief treatment (30 min) with the weak base methylamine, known to reverse the gap junction inhibitors effect, significantly (P < 0.05) reversed the inhibitory action of these agents on the blastocyst rate. Gap junction communication between the oocyte and surrounding cumulus cells was further studied using microinjection of the fluorescent dye Lucifer Yellow. Morphological evidences (dye transfer) were obtained that support the presence of functional coupling for a longer period with the FSH short exposure. In conclusion, high developmental rates of bovine oocytes can be achieved with a short exposure to r-hFSH. This effect is believed to be mediated through gap junctions as developmental competence of oocytes is compromised by the inhibition of their function. (c) 2005 Wiley-Liss, Inc.