Sydney Centre for Developmental and Regenerative Medicine, Sydney Medical School, University of Sydney, NSW, Australia.
Reprod Biol Endocrinol. 2010 Jun 23;8:71. doi: 10.1186/1477-7827-8-71.
This study investigates the suitability of surface-enhanced laser desorption and ionization time-of-flight (SELDI-TOF) and electrospray ionization (ESI) mass spectrometry for analysis of the proteins released by the mouse preimplantation embryo in vitro. SELDI-TOF analysis with CM10 or IMAC30 (but not Q10) protein chips detected a protein peak at m/z approximately 8570 released by both C57BL6 and hybrid embryos. No other peaks unique to the presence of the embryo were identified with this method. ESI mass spectrometry of tryptic digests of embryo-conditioned media identified a total of 20 proteins released during development from the zygote to blastocyst stage. Four proteins were expressed in at least 7 out of 8 cultures tested, one of these (lactate dehydrogenase B) was in all cultures. A further five proteins were in at least half of the cultures and 11 more proteins were in at least one culture. The expression of two of these proteins is essential for preimplantation embryo development (NLR family, pyrin domain containing 5 and peptidyl arginine deiminase, type VI). A further four proteins detected have roles in redox regulation of cells, and three others are capable of inducing post-translational modifications of proteins. This study shows the feasibility of ESI mass spectrometry for identifying the proteins secreted by the preimplantation embryo in vitro. This analysis identifies a range of targets that now require detailed functional analysis to assess whether their release by the embryo is an important property of early embryo development.
这项研究调查了表面增强激光解吸电离飞行时间（SELDI-TOF）和电喷雾电离（ESI）质谱法用于分析体外培养的小鼠着床前胚胎释放的蛋白质的适用性。CM10 或 IMAC30（但不是 Q10）蛋白芯片的 SELDI-TOF 分析检测到 C57BL6 和杂种胚胎都释放出约 8570m/z 的蛋白质峰。用这种方法没有发现其他与胚胎存在相关的独特峰。胚胎条件培养基的胰蛋白酶消化物的 ESI 质谱鉴定出总共 20 种在从受精卵到囊胚阶段发育过程中释放的蛋白质。在测试的 8 个培养物中至少有 7 个表达了 4 种蛋白质，其中一种（乳酸脱氢酶 B）存在于所有培养物中。另外 5 种蛋白质至少存在于一半的培养物中，还有 11 种蛋白质存在于至少一种培养物中。其中两种蛋白质的表达对于着床前胚胎的发育是必需的（NLR 家族，包含吡喃结构域的 5 型和肽酰精氨酸脱亚氨酶，6 型）。另外 4 种检测到的蛋白质在细胞的氧化还原调节中起作用，还有 3 种其他蛋白质能够诱导蛋白质的翻译后修饰。这项研究表明 ESI 质谱法用于鉴定体外培养的着床前胚胎分泌的蛋白质是可行的。这项分析确定了一系列的靶标，现在需要进行详细的功能分析，以评估胚胎释放这些靶标是否是早期胚胎发育的一个重要特性。
