Arts and Sciences
IRBIT (inositol-1,4,5-trisphosphate (IP3) receptors binding protein released with IP3) regulates fluid transport in the pancreatic duct and other epithelial cells. Most of the research done on IRBIT focuses on what transporters and channels IRBIT regulates. Much less is known about what regulates IRBIT and its distribution along the gastrointestinal tract. In the present study, we aim to determine the expression of IRBIT in the gastrointestinal (GI) tract and accessory glands such as the liver and pancreas. IRBIT has two isoforms- a short and a long. To date, antibodies are unable to differentiate between the two, so a different approach is required. PCR (polymerase chain reaction) with specific primers for each isoform will be used. We have harvested stomach, duodenum, jejunum, ileum, proximal and distal colon from male rats. Using a phosphate buffered saline solution containing zero calcium, we have selectively isolated the epithelial cells from each of the aforementioned segments of the GI tract. Using western blot, we found that IRBIT is expressed in duodenum, jejunum, ileum, proximal and distal colon. To test the expression of the short and long IRBIT, using a similar technique we have isolated epithelial cells from stomach, duodenum, jejunum, ileum, proximal and distal colon. Using Trizol-chloroform we have selectively isolated the total mRNA from each tissue. In future experiments, we will make a cDNA library from each tissue to perform PCR and determine the expression of short and long IRBIT.
Klosa, Payton; Dix, Marie; Bazaldua, Amanda; and Bouyer, Patrice, "Expression of IRBIT Along the Rat Gastrointestinal Tract" (2020). Symposium on Undergraduate Research and Creative Expression (SOURCE). 878.