INVITRO SYNTHESIS OF OVIDUCTAL SECRETORY PROTEINS BY ESTROGEN-TREATED OVARIECTOMIZED GILTS
INVITRO SYNTHESIS OF OVIDUCTAL SECRETORY PROTEINS BY ESTROGEN-TREATED OVARIECTOMIZED GILTS
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DOI：
10.1002/jez.1402620409
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发表时间：
1992-07-01
影响因子：
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通讯作者：
ALVAREZ, IM
中科院分区：
文献类型：
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作者：
BUHI, WC;ASHWORTH, CJ;ALVAREZ, IM
本研究的目的是鉴定、表征和检查卵巢切除（OVX）、玉米油（CO）-、雌激素（E）-、孕激素（P）-和E + P-处理的后备母猪的整个输卵管（WO）、壶腹（A）和峡部（I）组织从头合成的输卵管分泌蛋白（OSP）。在CO、E、P或E + P处理连续11天后，从OVX小母猪收集输卵管，并将组织与H-3-亮氨酸（H-3-leu）孵育。与CO-、P-或E + P处理的后备母猪相比，E-处理的WO外植体的H-3-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H-3-leu掺入不可透析大分子的速率更高（P < 0.05）。注意到位置的影响，A具有比WO或I更大（P < 0.01）的掺入率。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）和荧光法分析条件培养基。1D-SDS-PAGE分析显示WO和A中有三条主要的E依赖性条带（335，000、100，000和80，000 M（r）），I中有一条（335，000 M（r））。在A中发现的20，000 M（r）条带被E抑制，而在A中发现的60，000 M（r）条带被P诱导。（100，000 M（r））和3（80，000 M（r））分别转化为碱性和酸性100，000 M（r）蛋白和75，000 - 85，000 M（r）蛋白（pI < 4），但在I.在A中发现的碱性20，000 M（r）蛋白和酸性45，000 M（r）复合物被E.培养基的凝胶过滤显示了高M（r）部分（> 2 × 10（6）），该部分由E诱导，并且在A培养基中比I培养基中高6.8倍。这项研究清楚地表明：1）E处理的后备母猪的WO和A组织从头合成和分泌三种主要蛋白质（碱性100，000、酸性100，000和75，000 - 85，000 M（r）），2）这些E依赖蛋白在I或其它处理中未发现，3）由A合成的几种蛋白复合物被E处理抑制;和4）主要在A中产生的高M（r）级分被E诱导或扩增。
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 H-3-leucine (H-3-leu). Rates of H-3-leu incorporation into nondialyzable macromolecules by WO explants were greater (P < 0.01) with E- compared to CO-, P-, or E + P-treated gilts and greater (P < 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 < 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 < 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 (> 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.