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[在为放射自显影研究做准备过程中鸟类(虎皮鹦鹉,肖)松果体中吲哚的保留情况]

[Retention of indoles in the pineal organ of the bird (Melopsittacus undulatus, Shaw) during its preparation for radioautographic study].

作者信息

Balemans M, Juillard M T, Van Benthem J, Collin J P

出版信息

Biol Cell. 1983;49(3):257-66. doi: 10.1111/j.1768-322x.1984.tb00245.x.

Abstract

Since high-resolution radioautography (dipping technique) might be very useful for the study of indole metabolism in the pineal cells, the retention of [3H]-indoles has to be examined during the preparation of specimens for electron microscopy (EM). The pineal organ of the parakeet (Melopsittacus undulatus) was used in the present work. 1) Indole metabolism: following uptake of [3H]-5-hydroxytryptophan ([3H]-HW) in vivo and [3H]-5-hydroxytryptamine ([3H]-HT) in vitro in similar seasonal and nycthemeral conditions--all the known pineal indolic metabolites were recovered by thin layer chromatography. [3H]-5-methoxyindoles were also formed from [3H]-melatonin ([3H]-aMT). 2) The radioactivity of fluids used in the processing of pineal organs in EM was determined by liquid scintillation counting: (a) no exogenous [3H]-indoles could be revealed in EM solutions after [3H]-HW in vivo uptake. (b) 8.8 to 13.4% of [3H]-indoles were washed out by glutaraldehyde after [3H]-HT in vitro uptake. (c) most of the 5-methoxyindoles after in vitro uptake of [3H]-aMT were lost in glutaraldehyde. Our chromatography procedures did not permit the identification of [3H]-indoles extracted by the glutaraldehyde fixative. In previous experiments, [3H]-HW and [3H]-HT uptake showed the presence of selective radioautographic reactions in the cells of the receptor line; however, silver grains were scarce and diffusely distributed in the pineal parenchyma after [3H]-aMT uptake.

摘要

由于高分辨率放射自显影术(浸渍技术)可能对松果体细胞中吲哚代谢的研究非常有用,因此在制备用于电子显微镜(EM)的标本过程中,必须检查[3H] - 吲哚的保留情况。本研究使用了虎皮鹦鹉(Melopsittacus undulatus)的松果体器官。1)吲哚代谢:在相似的季节和昼夜条件下,体内摄取[3H] - 5 - 羟色氨酸([3H] - HW)和体外摄取[3H] - 5 - 羟色胺([3H] - HT)后,所有已知的松果体吲哚代谢产物均通过薄层色谱法回收。[3H] - 褪黑素([3H] - aMT)也可形成[3H] - 5 - 甲氧基吲哚。2)通过液体闪烁计数法测定用于松果体器官EM处理的液体的放射性:(a)体内摄取[3H] - HW后,EM溶液中未检测到外源[3H] - 吲哚。(b)体外摄取[3H] - HT后,8.8%至13.4%的[3H] - 吲哚被戊二醛洗脱。(c)体外摄取[3H] - aMT后,大部分5 - 甲氧基吲哚在戊二醛中丢失。我们的色谱程序无法鉴定戊二醛固定剂提取的[3H] - 吲哚。在先前的实验中,[3H] - HW和[3H] - HT摄取显示在受体系细胞中存在选择性放射自显影反应;然而,摄取[3H] - aMT后,银颗粒在松果体实质中稀少且分布分散。

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