BE-66 Pgp-1 deficiency alterations of intestinal developmental genes in C. elegans

Introduction/Background: Inflammatory Bowel Disease (IBD) is an autoimmune reaction causing chronic inflammation of the intestinal tract. The cause is unknown, and treatments are limited. A deficiency in P-glycoprotein (pgp), a protein expressed in intestinal epithelial cells, is linked to human IBD. We used pgp-deficient Caenorhabditis elegans (C. elegans) to investigate intestinal development without a p-glycoprotein molecule. Our laboratory has described several changes in intestinal function in pgp1-/- C. elegans. Here, we look to investigate the possible changes in gene expression leading to these changes. Hypothesis: We hypothesized pgp1-/- C. elegans would exhibit changes within genes used to control intestinal development, inhibiting effective development of the intestine. Methods: Control and pgp1-/- C. elegans were obtained from the Caenorhabditis Genetics Center at the University of Minnesota and cultured on Nematode Growth Medium. Age synchronization was done via bleaching of gravid adults to obtain embryos, which could grow to young adults. qRT-PCR was done using single worm RNA that was isolated and reverse transcribed into cDNA. Gene Expression was measured using TaqMan Gene Expression primer probe sets for skn1, elt2, end1, and end3. Results: Compared with controls, L1 aged pgp1-/- C. elegans had decreased expression of skn1 and increased expression of elt2 and end1. Adults showed increased expression of skn1. No changes were observed in L1 end3 or Adult elt2, end1, or end3. Conclusions: The changes in skn1 indicate changes in the early intestinal development program in C. elegans, reflected in downstream changes in elt2 and end1. These data suggest that pgp1-/- C. elegans have early changes in gene expression leading to previously described changes in the intestine.