Sensibar J A, Alger B, Tseng A, Berg L, Lee C
Dept. of Urology, Northwestern University School of Medicine, Chicago, IL 60611.
J Urol. 1990 Jan;143(1):161-6. doi: 10.1016/s0022-5347(17)39902-0.
Protein synthetic activities in the ventral prostate were assessed by two-dimensional electrophoresis in either four-day or seven-day castrated rats at different intervals following subcutaneous implantation of testosterone-filled silastic tubings for a period of up to four days. Prostatic tissues were cut into one to two mm. pieces and incubated in tissue culture medium containing S35-methionine (100 microCi/ml.) at 37C under 95% oxygen and 5% carbon dioxide for four hours. The incubated tissues were subjected to two-dimensional electrophoresis and radiofluorography. Analysis of protein spots detected in the fluorograms by computer-assisted densitometry revealed temporal changes in the synthesis of individual proteins by the ventral prostate of castrated rats following androgen treatment. Changes in two groups of proteins were evaluated: castration-induced proteins and androgen-dependent proteins. The level of synthesis of three castration-induced proteins (spots G, H, and I) declined rapidly upon testosterone treatment and reached a non-detectable level for spots G and H and a low level of synthesis for spot I by three days following androgen treatment. Synthesis of androgen-dependent proteins (spots D, E, and F) was activated by testosterone treatment. However, the time interval required to activate the synthesis of these proteins is different. Synthesis of protein spot D (prostatic binding protein) was detected as soon as half hour after the treatment. Synthesis of spots E and F, on the other hand, was not activated until 24 and 48 hours after the treatment, respectively. These changes in patterns of protein synthesis represent the characteristics of cellular responses to testosterone stimulation by the regressed prostate.
通过二维电泳法评估去势4天或7天的大鼠在皮下植入充满睾酮的硅橡胶管长达4天的不同时间间隔后腹侧前列腺中的蛋白质合成活性。将前列腺组织切成1至2毫米的小块,在含有S35 - 蛋氨酸(100微居里/毫升)的组织培养基中,于37℃、95%氧气和5%二氧化碳条件下孵育4小时。将孵育后的组织进行二维电泳和放射自显影。通过计算机辅助密度测定法分析放射自显影片中检测到的蛋白质斑点,揭示了雄激素处理后去势大鼠腹侧前列腺中单个蛋白质合成的时间变化。评估了两组蛋白质的变化:去势诱导蛋白和雄激素依赖性蛋白。三种去势诱导蛋白(斑点G、H和I)的合成水平在睾酮处理后迅速下降,在雄激素处理3天后,斑点G和H达到不可检测水平,斑点I的合成水平较低。雄激素依赖性蛋白(斑点D、E和F)的合成通过睾酮处理被激活。然而,激活这些蛋白质合成所需的时间间隔不同。处理后半小时即可检测到蛋白质斑点D(前列腺结合蛋白)的合成。另一方面,斑点E和F的合成分别直到处理后24小时和48小时才被激活。蛋白质合成模式的这些变化代表了退化前列腺对睾酮刺激的细胞反应特征。