Harney J P, Ali M, Vedeckis W V, Bazer F W
Animal Science Department, University of Florida, Gainesville 32611-0691.
Reprod Fertil Dev. 1994;6(2):211-9. doi: 10.1071/rd9940211.
Porcine conceptus secretory proteins were obtained from medium in which pig conceptuses, collected on Day 15 of pregnancy, were cultured for 30 h. Culture medium was pooled, dialysed and concentrated by Amicon ultrafiltration for retinol and retinoic acid (RA) binding studies. Proteins in the 20-kDa range, conceptus-secreted retinol-binding protein (RBP), bound both [3H]retinol and [3H]RA specifically. Cross-competition experiments indicate that [3H]RA was completely displaced with excess cold retinol; however, excess cold RA did not completely displace [3H]retinol, suggesting that conceptus RBP has greater affinity for retinol than RA. Cellular RBP and retinoic acid receptor (RAR)-alpha and RAR-gamma mRNA transcripts (0.7 kb; 3.8 and 2.8 kb; 3.4 kb respectively) were detected in poly (A)+ RNA isolated from Day-15 conceptus, Day-15 pregnant endometrium, late pregnant myometrium and late pregnant fetal tissues of pigs by Northern blot analysis. RAR-alpha and RAR-gamma immunoreactive proteins were detected in extracts of Day-15 conceptus, Day-15 pregnant endometrium and late pregnant fetal tissues by Western blot analysis. Collectively, results indicate that biochemical molecules required for retinoid transport, metabolism and regulatory effects are present in porcine conceptus and endometrial tissues during early pregnancy in swine.
猪胚胎分泌蛋白是从培养基中获得的，该培养基是将妊娠第15天收集的猪胚胎培养30小时后得到的。将培养基汇集、透析并通过Amicon超滤浓缩，用于视黄醇和视黄酸（RA）结合研究。20 kDa范围内的蛋白质，即胚胎分泌的视黄醇结合蛋白（RBP），能特异性结合[3H]视黄醇和[3H]RA。交叉竞争实验表明，过量的冷视黄醇能完全取代[3H]RA；然而，过量的冷RA并不能完全取代[3H]视黄醇，这表明胚胎RBP对视黄醇的亲和力比对RA的亲和力更强。通过Northern印迹分析，在从猪妊娠第15天的胚胎、妊娠第15天的子宫内膜、妊娠晚期的子宫肌层和妊娠晚期的胎儿组织中分离的聚腺苷酸（A）+RNA中检测到细胞RBP和视黄酸受体（RAR）-α和RAR-γmRNA转录本（分别为0.7 kb、3.8和2.8 kb、3.4 kb）。通过Western印迹分析，在妊娠第15天的胚胎、妊娠第15天的子宫内膜和妊娠晚期的胎儿组织提取物中检测到RAR-α和RAR-γ免疫反应蛋白。总体而言，结果表明，在猪早期妊娠期间，类视黄醇运输、代谢和调节作用所需的生化分子存在于猪胚胎和子宫内膜组织中。
