Identification of stem cells in small intestine and colon by marker gene Lgr5

The intestinal epithelium is the most rapidly self-renewing tissue in adult mammals. It is currently believed that four to six crypt stem cells reside at the +4 position immediately above the Paneth cells in the small intestine; colon stem cells remain undefined. Lgr5 (leucine-rich-repeat-containing G-protein-coupled receptor 5, also known as Gpr49) was selected from a panel of intestinal Wnt target genes for its restricted crypt expression. Here, using two knock-in alleles, we reveal exclusive expression of Lgr5 in cycling columnar cells at the crypt base. In addition, Lgr5 was expressed in rare cells in several other tissues. Using an inducible Cre knock-in allele and the Rosa26-lacZ reporter strain, lineage-tracing experiments were performed in adult mice. The Lgr5-positive crypt base columnar cell generated all epithelial lineages over a 60-day period, suggesting that it represents the stem cell of the small intestine and colon. The expression pattern of Lgr5 suggests that it marks stem cells in multiple adult tissues and cancers.     

The Effect of Worker Learning on Scheduling Jobs in a Hybrid Flow Shop: A Bi-Objective Approach

This paper studies learning effect as a resource utilization technique that can model improvement in worker’s ability as a result of repeating similar tasks. By considering learning of workers while performing setup times, a schedule can be determined to place jobs that share similar tools and fixtures next to each other. The purpose of this paper is to schedule a set of jobs in a hybrid flow shop (HFS) environment with learning effect while minimizing two objectives that are in conflict: namely maximum completion time (makespan) and total tardiness. Minimizing makespan is desirable from an internal efficiency viewpoint, but may result in individual jobs being scheduled past their due date, causing customer dissatisfaction and penalty costs. A bi-objective mixed integer programming model is developed, and the complexity of the developed bi-objective model is compared against the bi-criteria one through numerical examples. The effect of worker learning on the structure of assigned jobs to machines and their sequences is analyzed. Two solution methods based on the hybrid water flow like algorithm and non-dominated sorting and ranking concepts are proposed to solve the problem. The quality of the approximated sets of Pareto solutions is evaluated using several performance criteria. The results show that the proposed algorithms with learning effect perform well in reducing setup times and eliminate the need for setups itself through proper scheduling.     

Lgr5 homologues associate with Wnt receptors and mediate R-spondin signalling

The adult stem cell marker Lgr5 and its relative Lgr4 are often co-expressed in Wnt-driven proliferative compartments. We find that conditional deletion of both genes in the mouse gut impairs Wnt target gene expression and results in the rapid demise of intestinal crypts, thus phenocopying Wnt pathway inhibition. Mass spectrometry demonstrates that Lgr4 and Lgr5 associate with the Frizzled/Lrp Wnt receptor complex. Each of the four R-spondins, secreted Wnt pathway agonists, can bind to Lgr4, -5 and -6. In HEK293 cells, RSPO1 enhances canonical WNT signals initiated by WNT3A. Removal of LGR4 does not affect WNT3A signalling, but abrogates the RSPO1-mediated signal enhancement, a phenomenon rescued by re-expression of LGR4, -5 or -6. Genetic deletion of Lgr4/5 in mouse intestinal crypt cultures phenocopies withdrawal of Rspo1 and can be rescued by Wnt pathway activation. Lgr5 homologues are facultative Wnt receptor components that mediate Wnt signal enhancement by soluble R-spondin proteins. These results will guide future studies towards the application of R-spondins for regenerative purposes of tissues expressing Lgr5 homologues.