Boelhauve Marc, Sinowatz Fred, Wolf Eckhard, Paula-Lopes Fabíola F
Department of Molecular Animal Breeding and Biotechnology, Ludwig-Maximilian University, Oberschleissheim, Germany.
Biol Reprod. 2005 Oct;73(4):737-44. doi: 10.1095/biolreprod.105.041103. Epub 2005 Jun 15.
The series of events associated with oocyte growth and maturation determines the oocyte's ability to undergo successful fertilization, cleavage and embryonic development. Among the molecules involved in these events, leptin has been identified as a modulator of oocyte function. Experiments were conducted to determine whether leptin treatment of oocytes during maturation affects their developmental capacity after fertilization and whether it has long-lasting effects on apoptosis and gene expression in the resulting blastocysts. Cumulus-oocyte complexes (COCs) were matured in serum-free medium containing 0 (control), 1, 10, or 100 ng/ml leptin or in medium supplemented with 10% (v/v) estrous cow serum (ECS). Addition of leptin during oocyte maturation had no effect on cleavage rate after fertilization. However, an increased proportion of oocytes that matured in the presence of 1 or 10 ng/ml leptin developed to blastocysts, which exhibited increased cell numbers. The proportion of apoptotic cells was reduced in blastocysts originating from leptin- or ECS-treated oocytes. Transcript levels of the genes encoding leptin receptor (LEPR), signal transducer and activator of transcription (STAT3), BCL2 associated X-protein (BAX), and baculoviral inhibitor of apoptosis protein repeat-containing 4 (BIRC4, also known as XIAP), were determined by reverse transcriptase-quantitative polymerase chain reaction analysis of expanded and hatched blastocysts. Depending on the dose used, leptin treatment of oocytes resulted in increased LEPR, STAT3, and BIRC4 mRNA levels and reduced BAX mRNA levels in blastocysts. In conclusion, leptin improved the ability of the oocyte to sustain embryonic development and had long-term effects on blastocyst apoptosis and transcript abundance of LEPR, STAT3, and apoptosis-associated genes.
与卵母细胞生长和成熟相关的一系列事件决定了卵母细胞成功受精、分裂和胚胎发育的能力。在参与这些事件的分子中,瘦素已被确定为卵母细胞功能的调节因子。开展实验以确定在成熟过程中对卵母细胞进行瘦素处理是否会影响其受精后的发育能力,以及它是否对所得囊胚的凋亡和基因表达有长期影响。卵丘 - 卵母细胞复合体(COC)在含有0(对照)、1、10或100 ng/ml瘦素的无血清培养基中或在添加10%(v/v)发情母牛血清(ECS)的培养基中成熟。在卵母细胞成熟期间添加瘦素对受精后的分裂率没有影响。然而,在1或10 ng/ml瘦素存在下成熟的卵母细胞发育成囊胚的比例增加,且这些囊胚的细胞数量增多。来自经瘦素或ECS处理的卵母细胞的囊胚中凋亡细胞的比例降低。通过对扩张和孵化的囊胚进行逆转录 - 定量聚合酶链反应分析,测定了编码瘦素受体(LEPR)、信号转导和转录激活因子(STAT3)、BCL2相关X蛋白(BAX)以及含杆状病毒凋亡蛋白重复序列4(BIRC4,也称为XIAP)的基因的转录水平。根据所用剂量,对卵母细胞进行瘦素处理会导致囊胚中LEPR、STAT3和BIRC4的mRNA水平升高,而BAX的mRNA水平降低。总之,瘦素提高了卵母细胞维持胚胎发育的能力,并对囊胚凋亡以及LEPR、STAT3和凋亡相关基因的转录丰度有长期影响。