Buhi W C, Ashworth C J, Bazer F W, Alvarez I M
Department of Obstetrics and Gynecology, University of Florida, Gainesville 32610.
J Exp Zool. 1992 Jul 1;262(4):426-35. doi: 10.1002/jez.1402620409.
The objective of this study was to identify, characterize, and examine oviductal secretory proteins (OSP) synthesized de novo by whole oviduct (WO), ampulla (A), and isthmic (I) tissue from ovariectomized (OVX), corn oil (CO)-, estrogen (E)-, progesterone (P)-, and E + P-treated gilts. Oviducts were collected from OVX gilts after CO, E, P, or E + P treatment for 11 consecutive days and tissue was incubated with 3H-leucine (3H-leu). Rates of 3H-leu incorporation into nondialyzable macromolecules by WO explants were greater (P less than 0.01) with E- compared to CO-, P-, or E + P-treated gilts and greater (P less than 0.05) by A explants with E- compared to CO-, P-, or E + P-treated gilts. An effect of location was noted, with A having a greater (P less than 0.01) rate of incorporation than WO or I. Conditioned culture medium was analyzed by one (1D)- and two-dimensional (2D) sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and fluorography. Analyses by 1D-SDS-PAGE revealed three major E-dependent bands (335,000, 100,000, and 80,000 M(r)) in WO and A, and one (335,000 M(r)) in the I. A 20,000 M(r) band found in A was inhibited by E, while a 60,000 M(r) band found in the A was induced by P. Analyses by 2D-SDS-PAGE resolved major E-dependent bands 2 (100,000 M(r)) and 3 (80,000 M(r)) into basic and acidic 100,000 M(r) proteins and a 75,000-85,000 M(r) protein (pI less than 4), respectively, found in WO and A, but not in I. A basic 20,000 M(r) protein and an acidic 45,000 M(r) complex, both found in A, were inhibited by E. Gel filtration of culture medium revealed a high M(r) fraction (greater than 2 x 10(6)) that was induced by E and was 6.8-fold greater in medium from A than from I. This study clearly demonstrates that 1) WO and A tissue from E-treated gilts de novo synthesize and secrete three major proteins (basic 100,000, acidic 100,000, and 75,000-85,000 M(r)); 2) these E-dependent proteins are not found in I or with other treatment; 3) several protein complexes synthesized by A are inhibited by E treatment; and 4) a high M(r) fraction, produced primarily in the A, is induced or amplified by E.
本研究的目的是鉴定、表征并检测去卵巢（OVX）、经玉米油（CO）、雌激素（E）、孕酮（P）以及E+P处理的后备母猪的全输卵管（WO）、壶腹部（A）和峡部（I）组织新合成的输卵管分泌蛋白（OSP）。在CO、E、P或E+P连续处理11天后，从OVX后备母猪收集输卵管，组织与³H-亮氨酸（³H-leu）一起孵育。与CO、P或E+P处理的后备母猪相比，E处理的WO外植体将³H-leu掺入不可透析大分子的速率更高（P<0.01）；与CO、P或E+P处理的后备母猪相比，E处理的A外植体的掺入速率更高（P<0.05）。观察到位置的影响，A的掺入速率比WO或I更高（P<0.01）。通过一维（1D）和二维（2D）十二烷基硫酸钠（SDS）聚丙烯酰胺凝胶电泳（PAGE）和荧光自显影分析条件培养基。1D-SDS-PAGE分析显示，WO和A中有三条主要的E依赖性条带（相对分子质量分别为335,000、100,000和80,000），I中有一条（相对分子质量335,000）。在A中发现的相对分子质量20,000的条带受E抑制，而在A中发现的相对分子质量60,000的条带受P诱导。2D-SDS-PAGE分析将主要的E依赖性条带2（相对分子质量100,000）和条带3（相对分子质量80,000）分别解析为相对分子质量100,000的碱性和酸性蛋白以及相对分子质量75,000 - 85,000（等电点<4）的蛋白，这些蛋白存在于WO和A中，但不存在于I中。在A中发现的碱性相对分子质量20,000的蛋白和酸性相对分子质量45,000的复合物均受E抑制。培养基的凝胶过滤显示存在一个高相对分子质量组分（>2×10⁶），该组分受E诱导，且A培养基中的该组分比I培养基中的高6.8倍。本研究清楚地表明：1）E处理的后备母猪的WO和A组织新合成并分泌三种主要蛋白（碱性相对分子质量100,000、酸性相对分子质量100,000和相对分子质量75,0,00 - 85,000）；2）这些E依赖性蛋白在I中或其他处理组中未发现；3）A合成的几种蛋白复合物受E处理抑制；4）主要在A中产生的高相对分子质量组分受E诱导或放大。
