Teuscher Alina C, Statzer Cyril, Pantasis Sophia, Bordoli Mattia R, Ewald Collin Y
Department of Health Sciences and Technology, Institute of Translational Medicine, Eidgenössische Technische Hochschule (ETH) Zürich, Zürich, Switzerland.
Department of Biology, Institute of Molecular Health Sciences, Eidgenössische Technische Hochschule (ETH) Zürich, Zürich, Switzerland.
Methods Mol Biol. 2019;1944:169-188. doi: 10.1007/978-1-4939-9095-5_13.
Proper collagen homeostasis is essential for development and aging of any multicellular organism. During aging, two extreme scenarios are commonly occurring: a local excess in collagen deposition, for instance during fibrosis, or a gradual overall reduction of collagen mass. Here, we describe a histological and a colorimetric method to assess collagen levels in mammalian tissues and in the nematode Caenorhabditis elegans. The first method is the polychrome Herovici staining to distinguish between young and mature collagen ratios. The second method is based on hydroxyproline measurements to estimate collagen protein levels. In addition, we show how to decellularize the multicellular organism C. elegans in order to harvest its cuticle, one of the two major extracellular matrices, mainly composed of collagen. These methods allow assessing collagen deposition during aging either in tissues or in whole organisms.
适当的胶原蛋白稳态对于任何多细胞生物的发育和衰老至关重要。在衰老过程中，通常会出现两种极端情况：例如在纤维化过程中胶原蛋白沉积局部过量，或者胶原蛋白总量逐渐整体减少。在这里，我们描述了一种组织学方法和一种比色法，用于评估哺乳动物组织和线虫秀丽隐杆线虫中的胶原蛋白水平。第一种方法是多色赫罗维奇染色，用于区分年轻和成熟胶原蛋白的比例。第二种方法基于羟脯氨酸测量来估计胶原蛋白蛋白水平。此外，我们展示了如何使多细胞生物秀丽隐杆线虫脱细胞，以收获其角质层，角质层是两种主要细胞外基质之一，主要由胶原蛋白组成。这些方法可以评估衰老过程中组织或整个生物体中的胶原蛋白沉积情况。
