Goodwin Megan R, Rispoli Louisa A, Payton Rebecca R, Saxton Arnold M, Edwards J Lannett
University of Tennessee, Institute of Agriculture, UT AgResearch, Department of Animal Science, Knoxville, TN 37996-4574, USA.
J Reprod Dev. 2016 Dec 20;62(6):553-560. doi: 10.1262/jrd.2015-177. Epub 2016 Jul 21.
Because latent form of matrix metallopeptidase-9 (proMMP9) levels are positively related to blastocyst development, it was hypothesized that addition during maturation may improve development of heat-stressed oocytes. To test hypothesis, 0, 30 or 300 ng/ml human proMMP9 (hMMP9) was added at 18 h of in vitro maturation (hIVM) to cumulus-oocyte complexes matured at 38.5 or 41.0ºC (first 12 h only). Heat stress decreased 24 hIVM proMMP9 levels only in 0 and 30 ng/ml groups and increased progesterone in 0 and 300 ng/ml hMMP9 groups. Heat stress decreased cleavage and blastocyst development. Independent of maturation temperature, hMMP9 at 18 hIVM decreased blastocyst development. In a second study, cumulus-oocyte complexes were matured for 24 h at 38.5 or 41.0ºC (HS first 12 h only) with 0 or 300 ng/ml hMMP9 added at 12 hIVM. Without hMMP9, heat stress decreased 24 hIVM proMMP9 levels and increased progesterone production. Addition of 300 ng/ml of hMMP9 produced equivalent levels of proMMP9 at 24 hIVM (271 vs. 279 ± 77 for 38.5ºC and 41.0ºC treated oocytes, respectively). Heat stress did not affect ability of oocytes to cleave but reduced blastocyst development. Independent of temperature, hMMP9 decreased cleavage and blastocyst development. In summary, hMMP9 supplementation during IVM did not improve development of heat-stressed oocytes even when it was added for the entire maturation period. At doses tested, hMMP9 appeared detrimental to development when supplemented during the last 12 or 6 h of oocyte maturation.
由于基质金属蛋白酶-9（proMMP9）的潜伏形式水平与囊胚发育呈正相关，因此推测在成熟过程中添加可能会改善热应激卵母细胞的发育。为了验证这一假设，在体外成熟（hIVM）18小时时，向在38.5或41.0ºC（仅前12小时）成熟的卵丘-卵母细胞复合体中添加0、30或300 ng/ml的人proMMP9（hMMP9）。热应激仅在0和30 ng/ml组中降低了24小时hIVM proMMP9水平，并在0和300 ng/ml hMMP9组中增加了孕酮水平。热应激降低了卵裂率和囊胚发育率。与成熟温度无关，18小时hIVM时的hMMP9降低了囊胚发育率。在第二项研究中，卵丘-卵母细胞复合体在38.5或41.0ºC（仅热应激前12小时）成熟24小时，在hIVM 12小时时添加0或300 ng/ml hMMP9。在没有hMMP9的情况下，热应激降低了24小时hIVM proMMP9水平并增加了孕酮产量。添加300 ng/ml的hMMP9在24小时hIVM时产生了相当水平的proMMP9（38.5ºC和41.0ºC处理的卵母细胞分别为271 vs. 279±77）。热应激不影响卵母细胞的卵裂能力，但降低了囊胚发育率。与温度无关，hMMP9降低了卵裂率和囊胚发育率。总之，即使在整个成熟期间添加hMMP9，在IVM期间补充hMMP9也不能改善热应激卵母细胞的发育。在所测试的剂量下，在卵母细胞成熟的最后12或6小时补充hMMP9似乎对发育有害。
