Morphogenesis and Compartmentalization of the Intestinal Crypt.

The adult mammalian intestine is composed of two connected structures, the absorptive villi and the crypts, which house progenitor cells. Mouse crypts develop postnatally and are the architectural unit of the stem cell niche, yet the pathways that drive their formation are not known. Here, we combine transcriptomic, quantitative morphometric, and genetic analyses to identify mechanisms of crypt development. We uncover the upregulation of a contractility gene network at the earliest stage of crypt formation, which drives myosin II-dependent apical constriction and invagination of the crypt progenitor cells. Subsequently, hinges form, compartmentalizing crypts from villi. Hinges contain basally constricted cells, and this cell shape change was inhibited by increased hemidesmosomal adhesion in Rac1 null mice. Loss of hinges resulted in reduced villar spacing, revealing an unexpected role for crypts in tissue architecture and physiology. These studies provide a framework for studying crypt morphogenesis and identify essential regulators of niche formation.

Genetically induced microtubule disruption in the mouse intestine impairs intracellular organization and transport.

In most differentiated cells, microtubules reorganize into noncentrosomal arrays that are cell-type specific. In the columnar absorptive enterocytes of the intestine, microtubules form polarized apical-basal arrays that have been proposed to play multiple roles. However, in vivo testing of these hypotheses has been hampered by a lack of genetic tools to specifically perturb microtubules. Here we analyze mice in which microtubules are disrupted by conditional inducible expression of the microtubule-severing protein spastin. Spastin overexpression resulted in multiple cellular defects, including aberrations in nuclear and organelle positioning and deficient nutrient transport. However, cell shape, adhesion, and polarity remained intact, and mutant mice continued to thrive. Notably, the phenotypes of microtubule disruption are similar to those induced by microtubule disorganization upon loss of CAMSAP3/Nezha. These data demonstrate that enterocyte microtubules have important roles in organelle organization but are not essential for growth under homeostatic conditions.