Department of Animal Science, Division of Applied Life Science (BK21), Graduate School of Gyeongsang National University, Jinju, Republic of Korea.
Theriogenology. 2013 Jan 15;79(2):358-66.e1. doi: 10.1016/j.theriogenology.2012.10.008. Epub 2012 Nov 10.
It is unknown whether gene expression in cloned placenta during pre- and postimplantation is associated with early pregnancy failure in the cat. In this study, protein expression patterns were examined in early-stage (21-day-old) domestic cat placentas of fetuses derived from AI (CP; N = 4) and cloned embryo transfer (CEP; N = 2). Differentially expressed proteins were analyzed by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight (TOF) mass spectrometry (MS). A total of 21 proteins were aberrantly expressed (P < 0.05) by >1.5-fold in CEP compared with CP. Compared with CP, 12 proteins were upregulated in CEP (peptidyl-prolyl cis-trans isomerase A, annexin A2, protein DJ-1, adenylate kinase isoenzyme 1, protein disulfide-isomerase A3, actin cytoplasmic 1, serum albumin, protein disulfide-isomerase A6, and triosephosphate isomerase), and nine proteins were downregulated (triosephosphate isomerase; heterogeneous nuclear ribonucleoprotein H; tropomyosin alpha-4; triosephosphate isomerase 1; 60 kDa heat shock protein, mitochondrial; serum albumin; calumenin; keratin type 1; and prohibitin). The identities of the differentially expressed proteins were validated by peptide mass fingerprinting using matrix-assisted laser desorption/ionization-TOF/TOF MS/MS. The abnormally expressed proteins identified in this study might be associated with impaired development and dysfunction of CEP during early pregnancy. Abnormal protein expression might also induce fetal loss and contribute to failure to maintain pregnancy to term.
目前尚不清楚克隆胎盘在着床前和着床后的基因表达是否与猫的早期妊娠失败有关。在这项研究中,检查了来自人工授精(CP;N=4)和克隆胚胎转移(CEP;N=2)的胎儿的早期(21 天大)家猫胎盘的蛋白质表达模式。通过二维凝胶电泳和基质辅助激光解吸/电离飞行时间(TOF)质谱(MS)分析差异表达的蛋白质。与 CP 相比,CEP 中共有 21 种蛋白质的表达异常(P<0.05),差异超过 1.5 倍。与 CP 相比,CEP 中有 12 种蛋白质上调(肽基脯氨酰顺反异构酶 A、膜联蛋白 A2、DJ-1 蛋白、腺苷酸激酶同工酶 1、蛋白二硫键异构酶 A3、肌动蛋白细胞质 1、血清白蛋白、蛋白二硫键异构酶 A6 和磷酸丙糖异构酶),而 9 种蛋白质下调(磷酸丙糖异构酶;异质核核糖核蛋白 H;α-4 原肌球蛋白;磷酸丙糖异构酶 1;60kDa 热休克蛋白,线粒体;血清白蛋白;钙网蛋白;角蛋白 1;抑制素)。使用基质辅助激光解吸/电离-TOF/TOF MS/MS 通过肽质量指纹图谱验证了差异表达蛋白的身份。本研究中鉴定的异常表达蛋白可能与早期妊娠期间 CEP 的发育受损和功能障碍有关。异常的蛋白质表达也可能导致胎儿丢失,并导致妊娠无法维持到足月。