Vallet J L, Christenson R K, Klemcke H G
USDA, ARS, Roman L. Hruska U.S. Meat Animal Research Center, Clay Center, NE 68933, USA.
J Anim Sci. 1999 May;77(5):1236-40. doi: 10.2527/1999.7751236x.
A RIA was developed for porcine intrauterine folate binding protein (FBP). Displacement of [125I]FBP caused by increasing dilutions of uterine flushings collected from either d-15 pregnant or nonpregnant gilts or media from culture of endometrial tissue from d-15 pregnant or nonpregnant gilts was parallel to the displacement caused by the standard curve. Addition of known amounts of purified allantoic fluid FBP to dilutions of either intrauterine flushings or endometrial culture medium were measured accurately with the RIA. To test specificity, 2-mL samples of uterine flushings collected from d-15 pregnant and nonpregnant gilts were preincubated with 10 microCi of [3H]folic acid and then chromatographed using Sephadex G-100 (Sigma Chemical Co., St. Louis, MO). The fractions were subsequently assayed for radioactivity by liquid scintillation spectrophotometry and for FBP by RIA. The [3H]folic acid and FBP peaks coincided, indicating that the RIA is specific for FBP. Uterine flushings were collected on d 10, 11, 12, 13, 14, and 15 of the cycle or pregnancy from 1) White crossbred, 2) progesterone-treated White crossbred (200 mg of progesterone at 48 and 72 h after estrus), and 3) Meishan gilts and assayed for FBP. Total FBP increased 140-fold from d 10 to 15, and the pattern of change across day did not differ between pregnant and nonpregnant gilts. Progesterone treatment increased intrauterine FBP content on d 10 and 11. No difference in FBP concentrations was detected between White crossbred and Meishan gilts. These results indicate that the RIA for FBP is valid, allowing measurement of this protein in uterine flushings and endometrial culture medium. The onset of FBP secretion by the uterus between d 10 and 15 of the cycle or pregnancy is influenced by the timing of onset of progesterone influence in a manner similar to the endometrial proteins uteroferrin and retinol binding protein. In contrast to these endometrial proteins, FBP concentrations are similar in Meishan and White crossbred gilts.
已开发出一种用于检测猪子宫内叶酸结合蛋白（FBP）的放射免疫分析方法。从妊娠第15天的妊娠母猪或未妊娠母猪收集的子宫冲洗液稀释液，或从妊娠第15天的妊娠母猪或未妊娠母猪的子宫内膜组织培养物培养基中，[125I]FBP的取代情况与标准曲线引起的取代情况平行。用放射免疫分析法准确测定了向子宫冲洗液或子宫内膜培养基稀释液中添加已知量的纯化尿囊液FBP后的情况。为了测试特异性，将从妊娠第15天的妊娠母猪和未妊娠母猪收集的2 mL子宫冲洗液样品与10微居里的[3H]叶酸预孵育，然后使用Sephadex G - 100（西格玛化学公司，密苏里州圣路易斯）进行色谱分析。随后通过液体闪烁分光光度法测定各馏分的放射性，并通过放射免疫分析法测定FBP。[3H]叶酸峰和FBP峰重合，表明该放射免疫分析法对FBP具有特异性。在发情周期或妊娠期的第10、11、12、13、14和15天收集以下母猪的子宫冲洗液并检测FBP：1）白色杂交母猪；2）用孕酮处理的白色杂交母猪（发情后48小时和72小时各注射200毫克孕酮）；3）梅山母猪。总FBP从第10天到第15天增加了140倍，妊娠母猪和未妊娠母猪之间每天的变化模式没有差异。孕酮处理使第10天和第11天子宫内FBP含量增加。未检测到白色杂交母猪和梅山母猪之间FBP浓度的差异。这些结果表明，用于FBP的放射免疫分析法是有效的，能够测定子宫冲洗液和子宫内膜培养基中的这种蛋白质。发情周期或妊娠期第10天至第15天子宫分泌FBP的起始受到孕酮影响起始时间的影响，其方式类似于子宫内膜蛋白子宫铁蛋白和视黄醇结合蛋白。与这些子宫内膜蛋白不同的是，梅山母猪和白色杂交母猪的FBP浓度相似。
