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芳香化酶免疫反应性特异性定位于发育中小鼠下丘脑和皮质的神经元中。

Aromatase-immunoreactivity is localised specifically in neurones in the developing mouse hypothalamus and cortex.

作者信息

Beyer C, Green S J, Barker P J, Huskisson N S, Hutchison J B

机构信息

MRC Neuroendocrine Development and Behaviour Group, BABRAHAM Institute, Cambridge, UK.

出版信息

Brain Res. 1994 Feb 28;638(1-2):203-10. doi: 10.1016/0006-8993(94)90651-3.

Abstract

Local formation of oestrogens from androgens by aromatase cytochrome P-450 within brain cells is crucial for the sexual differentiation of the mammalian CNS. Aromatase activity has been detected in several brain regions of the developing rodent brain. In the present study, we used a mouse-specific, peptide-generated, polyclonal aromatase antibody to determine whether neurones and/or glial cells in the developing brain are involved in androgen aromatization and if aromatase-immunoreactive (Arom-IR) cells exhibit a sex-specific distribution and regional-specific morphological characteristics. For these experiments, gender-specific cell cultures were prepared from embryonic day 15 mouse hypothalamus and cortex. Specificity of the immunoreaction was confirmed by Western-blot analysis and by inhibition of aromatase activity using tissue homogenates from mouse ovaries and male newborn hypothalamus and from male hypothalamic cultures with known aromatase activity, respectively. Arom-IR cells were found in both hypothalamic and cortical cultures. Double-labeling experiments revealed that Arom-IR cells co-stained only for the neuronal marker MAP II, but never for glial markers. Therefore aromatase immunoreactivity is specifically neuronal. Regional differences in the morphology of Arom-IR neurones were observed between both brain regions. In hypothalamic cultures, IR-neurones represented a heterologous population of phenotypes (magnocellular, small bipolar and multipolar neurones with long processes showing varicose-like structures or without processes). Cortical Arom-IR neurones were always oval in shape with short or no IR-processes. Sexual dimorphisms in numbers of Arom-IR neurones were found in the hypothalamus with significantly higher cell numbers in male cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

脑细胞内的细胞色素P-450芳香化酶将雄激素转化为雌激素的局部过程,对哺乳动物中枢神经系统的性别分化至关重要。在发育中的啮齿动物大脑的几个区域已检测到芳香化酶活性。在本研究中,我们使用一种小鼠特异性的、肽段产生的多克隆芳香化酶抗体,以确定发育中的大脑中的神经元和/或神经胶质细胞是否参与雄激素的芳香化过程,以及芳香化酶免疫反应性(Arom-IR)细胞是否表现出性别特异性分布和区域特异性形态特征。对于这些实验,从胚胎第15天的小鼠下丘脑和皮质制备了性别特异性细胞培养物。免疫反应的特异性分别通过蛋白质免疫印迹分析以及使用来自小鼠卵巢、雄性新生鼠下丘脑的组织匀浆和具有已知芳香化酶活性的雄性下丘脑培养物抑制芳香化酶活性来证实。在下丘脑和皮质培养物中均发现了Arom-IR细胞。双重标记实验表明,Arom-IR细胞仅与神经元标记物微管相关蛋白2(MAP II)共染色,而从不与神经胶质标记物共染色。因此,芳香化酶免疫反应性具有特异性的神经元特征。在两个脑区之间观察到Arom-IR神经元形态的区域差异。在下丘脑培养物中,免疫反应性神经元代表了异源的表型群体(大细胞、小双极和多极神经元,具有长突起,显示出静脉曲张样结构或无突起)。皮质Arom-IR神经元总是呈椭圆形,突起短或无突起。在下丘脑中发现Arom-IR神经元数量存在性别二态性,雄性培养物中的细胞数量明显更多。(摘要截短于250字)

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