Fluorescence properties and staining behavior of monodansylpentane, a structural homologue of the lysosomotropic agent monodansylcadaverine
Fluorescence properties and staining behavior of monodansylpentane, a structural homologue of the lysosomotropic agent monodansylcadaverine
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DOI：
10.1177/002215540104900205
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发表时间：
2001-02-01
影响因子：
3.2
通讯作者：
Els채sser, HP
中科院分区：
文献类型：
--
作者：
Niemann, A;Baltes, J;Els채sser, HP
我们最近发现，单胺尸体碱可以标记自噬液泡。对标记机制的分析表明，单药尸胺作为溶酶性体剂，通过离子捕获机制被浓缩到酸性隔间中，作为溶剂极性探针，通过与高度集中在自噬液泡中的膜脂相互作用，增加其相对荧光强度。在本研究中，我们合成了三种结构上相关的单胺甲鼠碱衍生物，用中性基（dansylamylamine； MDH）、极性基（dansylaminopentanol； MDOH）或酸性基（danslaminovaleric acid； MDA）取代单胺甲鼠碱的一级氨基，以取代标记物的溶酶性。MDH对自噬液泡有特异性的染色，而极性和酸性衍生物则没有任何染色。我们进一步证明，自噬液泡的MDH染色不依赖于酸性pH值，因此不依赖于离子捕获机制，但它仍然显示出与单氨尸胺相同的自噬膜脂的偏好。我们提出MDH可以特异性地与自噬型板层溶酶体标志物相互作用，作为溶剂极性探针。因此，丹化氨基戊烷可以作为体内和固定细胞中自噬液泡的特异性标记物。
We have recently shown that monodansylcadaverine labels autophagic vacuoles. Analysis of the mechanism underlying the labeling revealed that monodansylcadaverine acts as a lysosomotropic agent, being concentrated into acidic compartments by an ion-trapping mechanism, and as a solvent polarity probe, increasing its relative fluorescence intensity by interacting with membrane lipids that are highly concentrated in the autophagic vacuoles. In this study, we synthesized three structurally related derivatives of monodansylcadaverine, replacing the primary amino group of monodansylcadaverine with a neutral (dansylamylamine; MDH), a polar (dansylaminopentanol; MDOH), or an acidic group (danslaminovaleric acid; MDA), to replace the lysosomotropic character of the marker. Whereas MDH showed a specific staining of autophagic vacuoles, the polar and acidic derivatives did not show any staining. We further demonstrate that the MDH staining of autophagic vacuoles is independent on the acidic pH and thus on an ion-trapping mechanism, but it still shows the same preferences for autophagic membrane lipids as monodansylcadaverine. We propose that MDH can specifically interact with lamellar bod lysosomal marker ies of the autophagic type as a solvent polarity probe. Therefore, dansylated aminopentane can be used as a specific marker for autophagic vacuoles in vivo and in fixed cells.