Tan Chin Wee, Hirokawa Yumiko, Gardiner Bruce S, Smith David W, Burgess Antony W
The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia ; Department of Medical Biology, University of Melbourne, Parkville, Victoria, Australia ; Ludwig Institute for Cancer Research, Melbourne-Parkville Branch, Parkville, Victoria, Australia.
PLoS One. 2013 Oct 21;8(10):e78519. doi: 10.1371/journal.pone.0078519. eCollection 2013.
The process of crypt formation and the roles of Wnt and cell-cell adhesion signaling in cryptogenesis are not well described; but are important to the understanding of both normal and cancer colon crypt biology. A quantitative 3D-microscopy and image analysis technique is used to study the frequency, morphology and molecular topography associated with crypt formation. Measurements along the colon reveal the details of crypt formation and some key underlying biochemical signals regulating normal colon biology. Our measurements revealed an asymmetrical crypt budding process, contrary to the previously reported symmetrical fission of crypts. 3D immunofluorescence analyses reveals heterogeneity in the subcellular distribution of E-cadherin and β-catenin in distinct crypt populations. This heterogeneity was also found in asymmetrical budding crypts. Singular crypt formation (i.e. no multiple new crypts forming from one parent crypt) were observed in crypts isolated from the normal colon mucosa, suggestive of a singular constraint mechanism to prevent aberrant crypt production. The technique presented improves our understanding of cryptogenesis and suggests that excess colon crypt formation occurs when Wnt signaling is perturbed (e.g. by truncation of adenomatous polyposis coli, APC protein) in most colon cancers.
隐窝形成的过程以及Wnt和细胞间黏附信号在隐窝发生中的作用尚未得到充分描述；但对于理解正常和癌性结肠隐窝生物学都很重要。一种定量三维显微镜和图像分析技术被用于研究与隐窝形成相关的频率、形态和分子拓扑结构。沿结肠的测量揭示了隐窝形成的细节以及一些调节正常结肠生物学的关键潜在生化信号。我们的测量揭示了一种不对称的隐窝出芽过程，这与先前报道的隐窝对称分裂相反。三维免疫荧光分析揭示了不同隐窝群体中E-钙黏蛋白和β-连环蛋白亚细胞分布的异质性。这种异质性在不对称出芽的隐窝中也被发现。在从正常结肠黏膜分离的隐窝中观察到单个隐窝形成（即没有一个亲代隐窝形成多个新隐窝），提示存在一种单一的约束机制来防止异常隐窝产生。所提出的技术增进了我们对隐窝发生的理解，并表明在大多数结肠癌中，当Wnt信号受到干扰（例如通过截短腺瘤性息肉病基因，APC蛋白）时会发生过多的结肠隐窝形成。
