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一种通过定量使用法尔克-希拉尔普方法获得的多巴胺荧光来测定离散多巴胺神经末梢系统中多巴胺水平和周转率的方法。

A method to determine dopamine levels and turnover rate in discrete dopamine nerve terminal systems by quantitative use of dopamine fluorescence obtained by Falck--Hillarp methodology.

作者信息

Agnati L F, Andersson K, Wiesel F, Fuxe K

出版信息

J Neurosci Methods. 1979 Dec;1(4):365-73. doi: 10.1016/0165-0270(79)90025-6.

Abstract

In tissue sections of regions with evenly distributed dopamine (DA) nerve terminals such as the nucleus caudatus, it is possible to obtain absolute amounts (nmol/g) of DA by means of quantitative microfluorimetrical measurements of catecholamine (CA) fluorescence intensity in the tissue and in DA-containing albumin-agar standards. The values agree well with mass-fragmentographical determinations of DA. The steady-state DA levels of heterogenously innervated areas such as the tuberculum olfactorium determined by quantitative microfluorimetry can be made comparable to the steady-state levels of DA obtained by biochemistry in the entire tuberculum olfactorium by means of a conversion factor which considers the dilution of the DA-containing structures by non-DA-containing nerve cells in the biochemical analysis. This factor is obtained by making biochemical determinations in such regions of untreated animals (e.g. the tuberculum olfactorium). The results of this paper have demonstrated that the histochemical approach not only has a high power of resolution and makes it possible to perform studies on an intact morphological substrate but also allows the determination of DA steady-state (nmol/g) and DA turnover rate (nmol/g x min-1) in discrete DA nerve terminal systems of the brain in absolute amounts.

摘要

在诸如尾状核等多巴胺(DA)神经末梢分布均匀的区域的组织切片中,通过对组织和含DA的白蛋白 - 琼脂标准品中儿茶酚胺(CA)荧光强度进行定量显微荧光测量,可以获得DA的绝对量(nmol/g)。这些值与DA的质量碎片分析法测定结果非常吻合。通过定量显微荧光法测定的诸如嗅结节等神经支配不均匀区域的稳态DA水平,可以通过一个转换因子与通过生物化学方法在整个嗅结节中获得的DA稳态水平进行比较,该转换因子考虑了生物化学分析中含DA结构被不含DA的神经细胞稀释的情况。这个因子是通过对未处理动物的此类区域(如嗅结节)进行生物化学测定而获得的。本文的结果表明,组织化学方法不仅具有高分辨率,能够在完整的形态学底物上进行研究,而且还能以绝对量测定大脑离散DA神经末梢系统中的DA稳态(nmol/g)和DA周转率(nmol/g×min⁻¹)。

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