Vázquez Cristina Lourdes, Colombo María Isabel
Laboratorio de Biología Celular y Molecular, Instituto de Histología y Embriología (IHEM), Facultad de Ciencias Médicas, Universidad Nacional de Cuyo-CONICET, Mendoza, Argentina.
Methods Enzymol. 2009;452:85-95. doi: 10.1016/S0076-6879(08)03606-9.
In this chapter we describe the use of monodasylcadaverine (MDC) and DQ-BSA, two practical and convenient tools to study the autophagic pathway. MDC is a lysosomotropic compound useful for the identification of autophagic vesicles by fluorescence microscopy and, in addition, to assess autophagy induction via the accumulation of MDC-labeled vacuoles. However, the increase of autophagosomes does not necessarily reflect autophagosome maturation and degradation of the sequestered materials, thus the use of DQ-BSA in conjunction with and autophagic marker is an appropriate technique to monitor the formation of the autolysosome, the degradative compartment. Therefore, here we discuss the advantages of the utilization of these two methods to characterize the autophagy pathway.
在本章中，我们描述了单丹磺酰尸胺（MDC）和DQ-牛血清白蛋白（DQ-BSA）这两种实用且便捷的工具在自噬途径研究中的应用。MDC是一种溶酶体亲和性化合物，可通过荧光显微镜用于鉴定自噬泡，此外，还可通过MDC标记的液泡积累来评估自噬诱导。然而，自噬体的增加并不一定反映自噬体的成熟以及被隔离物质的降解，因此，将DQ-BSA与自噬标记物结合使用是监测自噬溶酶体（即降解区室）形成的合适技术。所以，在此我们讨论使用这两种方法来表征自噬途径的优势。
