Immunocytochemical studies of aromatase in early and full-term human placental tissues: comparison with biochemical assays.
Immunocytochemical studies of aromatase in early and full-term human placental tissues: comparison with biochemical assays.
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早期和足月人胎盘组织中芳香酶的免疫细胞化学研究：与生化测定的比较。
DOI：
10.1095/biolreprod41.5.889
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发表时间：
1989
影响因子：
3.6
通讯作者：
Brodie,AM
中科院分区：
文献类型：
--
作者：
Inkster,SE;Brodie,AM
利用特异性单克隆抗体可视化芳香化酶p450酶的免疫细胞化学方法已被开发出来，用于未固定的冷冻组织切片。我们比较了单克隆和多克隆芳香化酶特异性抗体，发现胎盘芳香化酶一致且专门位于绒毛膜绒毛的合胞滋养层。该单克隆抗体具有最高亲和力，相关背景染色可忽略不计。发现固定物损害染色反应。检查早期妊娠和足月胎盘显示相同的免疫染色模式相似的强度在10个样本中的9个。以[1,2,6,7−3H]雄烯二酮为底物，采用间接放射性氚水（3H2O）法和高效液相色谱（HPLC）法同时测定了孕早期和足月胎盘的免疫染色反应，比较了它们各自的微粒体芳香酶活性。发现这两种测定法对足月胎盘标本的酶活性估计具有可比性。然而，当分析孕早期标本时，直接产物测量值明显大于相应的3h2o测定结果。然而，发现生化芳香酶活性与免疫染色反应呈正相关。虽然没有直接测量17β-羟基类固醇脱氢酶的活性，但雌二醇：雌二醇产物比的差异（妊娠前期2.49±0.68 vs.妊娠期0.89±0.15）表明该酶在妊娠的两个阶段受到不同的控制。一个早期妊娠标本具有不典型，斑片免疫染色分布也具有极低的芳香酶活性。结果表明，这两种抗体都能识别功能性芳香酶，并表明免疫细胞化学检测是一种敏感的、定性的技术来研究这种重要的甾体生成酶。
An immunocytochemical method for visualizing the aromatase P450enzyme with a specific monoclonal antibody has been developed for use with unfixed, frozen tissue sections. We compared both monoclonal and polyclonal aromatase-specific antibodies and found that placental aromatase was consistently and exclusively located in the syncytiotrophoblast layer of chorionic villi. The monoclonal antibody had the highest affinity, with negligible associated background stain. Fixation was found to impair stain reaction. Examination of first trimester and term placentae revealed identical immunostaining patterns of similar intensity in 9 of 10 samples.The immunostain reactions of first trimester and term placentae were compared with their respective microsomal aromatase activity, determined simultaneously by both indirect radiometric tritiated water (3H2O) assay, and direct product isolation by HPLC, using [1, 2, 6, 7−3H] androstenedione as substrate. The two assays were found to be comparable for enzyme activity estimates of term placental specimens. However, when first trimester specimens were analyzed, the direct-product measurements were significantly larger than the corresponding3H2O assay results. Nonetheless, biochemical aromatase activity was found to correlate positively with immunostain reaction. Although 17β-hydroxysteroid dehydrogenase activity was not directly measured, differences in the estradiol:estrone product ratio (2.49 ± 0.68 first trimester vs. 0.89 ± 0.15 term) suggest differential control of this enzyme at the two stages of pregnancy. One first trimester specimen with an atypical, patchy immunostain distribution also had extremely low aromatase activity. The results indicate that both antibodies recognize functional aromatase enzyme and suggest that immunocytochemical detection is a sensitive, qualitative technique for investigating this important steroidogenic enzyme.
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一些啮齿动物、牛和人类卵巢中芳香酶的免疫细胞化学定位
DOI：
--
发表时间：
1984
期刊：
影响因子：
--
作者：
H. Matsuda;H. Fujita;K. Ishimura;Y. Osawa
通讯作者：
Y. Osawa
大鼠卵巢雌激素生物合成机制的研究--Ⅰ．
DOI：
--
发表时间：
1976
期刊：
Journal of Steroid Biochemistry
影响因子：
--
作者：
A. Brodie;W. C. Schwarzel;H. Brodie
通讯作者：
H. Brodie
人皮肤成纤维细胞中的芳香酶活性：通过酶法表征。
DOI：
--
发表时间：
1986
期刊：
Journal of Steroid Biochemistry
影响因子：
--
作者：
A. Chabab;J. Nicolas;C. Sultan
通讯作者：
C. Sultan
通过亲和层析纯化人胎盘雌激素合成酶（芳香酶）。
DOI：
10.1016/s0021-9258(18)61668-0
发表时间：
1987
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
D. Hagerman
通讯作者：
D. Hagerman
与人胎盘线粒体部分相关的芳香酶系统的特性
影响因子：
2.7
作者：
J. Canick;K. Ryan
通讯作者：
K. Ryan