Cholecystokinin (CCK) secreting enteroendocrine (EEC) I cells which distribute in gastrointestinal tract play an important role in lipid sensing, digestion and fatty acids uptake. Although a lot of research has been performed, the whole mechanism of fat sensing and fatty acid uptake and hormone expression in the CCK cells is still unclear. Global analysis to characterise the CCK cells is essential. CCK cells have an indistinct morphology, a diffuse distribution and a small percentage of population in the small intestine. However, the generation of genetic fluorescence tagged animal model facilitates the study of these cells. In this thesis, single cell dissociation methods and RT-PCR methodologies for detecting nutrient sensing receptors and fatty acids transporters were established and optimised. Expression of mRNAs for fat sensing GPCRs were detected in mouse duodenal epithelium. Expression of FATP family and CD36 in CCK cells and enterocytes was studied by RT-PCR. FATP2, FATP4 and CD36 mRNA were found in both CCK cells and enterocytes. Cell culture methodologies enabling the study of function (calcium imaging and FACS analysis) were established and optimised by checking the cell viability as a criterion. The methodology combining the immunochemistry and FACS analysis to study the hormone was established but requires further optimisation.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:756788 |
| Date | January 2014 |
| Creators | Huang, Xiaoxing |
| Contributors | Smith, Craig ; Mclaughlin, John |
| Publisher | University of Manchester |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | https://www.research.manchester.ac.uk/portal/en/theses/characterisation-of-mouse-duodenal-cholecystokinin-cell(3db78a29-21a7-4b1a-9664-f4d0f493f90b).html |
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