17beta-Estradiol levels in male zebra finch brain: combining Palkovits punch and an ultrasensitive radioimmunoassay.
17beta-Estradiol levels in male zebra finch brain: combining Palkovits punch and an ultrasensitive radioimmunoassay.
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DOI：
10.1016/j.ygcen.2010.02.002
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发表时间：
2010-05-15
影响因子：
2.7
通讯作者：
Soma, Kiran K.
中科院分区：
文献类型：
--
作者：
Charlier, Thierry D.;Po, Kelvin W. L.;Newman, Amy E. M.;Shah, Amit H.;Saldanha, Colin J.;Soma, Kiran K.
关键词：
睾酮的生理和行为作用通常需要睾酮局部芳构化为17β-雌二醇（E2）。在大多数脊椎动物中，芳香化酶在少数离散的脑区域中表达。虽然许多研究已经测量了大脑芳香化酶mRNA或活性，但由于技术挑战，很少有研究测量大脑E2水平，特别是在离散的大脑区域。在这里，我们使用的Palkovits打孔技术，以分离13个离散的脑细胞核成年雄性斑胸草雀。通过固相萃取法提取类固醇。然后用超灵敏、特异和精确的放射免疫测定法测定E2。我们的方案导致E2的高回收率（84%），并有效地去除干扰脑脂质。E2水平高芳香化酶丰富的地区，如尾内侧巢皮质和海马。内侧视前区、下丘脑腹内侧核、前巢皮质外侧和内侧大细胞核、杏仁核带核和X区的E2水平居中。E2水平在小脑、HVC、外侧巢皮质和视叶中基本上检测不到。重要的是，血浆中的E2水平显著低于尾侧内侧巢皮质。该协议允许一个测量E2在离散的大脑区域和潜在的相关的局部E2浓度芳香化酶的活动和行为。
Local aromatization of testosterone into 17β-estradiol (E2) is often required for the physiological and behavioral actions of testosterone. In most vertebrates, aromatase is expressed in a few discrete brain regions. While many studies have measured brain aromatase mRNA or activity, very few studies have measured brain E2 levels, particularly in discrete brain regions, because of technical challenges. Here, we used the Palkovits punch technique to isolate 13 discrete brain nuclei from adult male zebra finches. Steroids were extracted via solid phase extraction. E2 was then measured with an ultrasensitive, specific and precise radioimmunoassay. Our protocol leads to high recovery of E2 (84%) and effectively removes interfering brain lipids. E2 levels were high in aromatase-rich regions such as caudal medial nidopallium and hippocampus. E2 levels were intermediate in the medial preoptic area, ventromedial nucleus of the hypothalamus, lateral and medial magnocellular nuclei of anterior nidopallium, nucleus taeniae of the amygdala, and Area X. E2 levels were largely non-detectable in the cerebellum, HVC, lateral nidopallium and optic lobes. Importantly, E2 levels were significantly lower in plasma than in the caudal medial nidopallium. This protocol allows one to measure E2 in discrete brain regions and potentially relate local E2 concentrations to aromatase activity and behavior.
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