Enteroendocrine cell expression of a cholecystokinin gene construct in transgenic mice and cultured cells

Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, Michigan, United States
AJP Gastrointestinal and Liver Physiology (Impact Factor: 3.8). 03/2005; 288(2):G354-61. DOI: 10.1152/ajpgi.00356.2004
Source: PubMed


CCK is predominantly expressed in subsets of endocrine cells in the intestine and neurons in the brain. We evaluated the expression of a CCK gene construct in transgenic mice and cultured cells to identify a genomic region that directs correct tissue- and cell-specific expression in enteroendocrine cells. The CCKL1 transgene contained 6.4 kb of mouse Cck fused to lacZ. Expression was evaluated in three transgenic lines (J11, J12, J14) by measurement of beta-galactosidase in tissue homogenates and frozen sections. Correct tissue-specific expression was observed, with beta-galactosidase activity detected in intestine and brain. However, there were differences seen in cell-specific expression in the intestine. Line J14 exhibited expression in CCK-endocrine cells, with expressing cells arising at the normal time during fetal development. However, transgene expression in line J12 intestine was limited to neurons of the enteric nervous system, which reflect an early fetal expression pattern for CCK. Analysis of an additional 15 transgenic founder mice demonstrated intestinal expression in 40% of transgenics, with expressing mice following either an endocrine cell pattern or a neuronal pattern in approximately equal numbers. CCKL1 transfection analysis in cultured cells also demonstrated enteroendocrine cell expression, with 100-fold enhanced activity in the enteroendocrine cell line STC-1 compared with nonendocrine cell lines. The results suggest that the minimal cis-regulatory DNA elements necessary for appropriate CCK expression in enteroendocrine cells reside within the 6.4-kb mouse genomic fragment.

    • "However, when in situ hybridization was coupled with immunocytochemistry, not all the CCK-positive cells were labelled with the DsRedT3 antisense riboprobe (Fig. 2b), indicating that the transgene expression is highly specific, but not complete. Several studies on transgenic mice have reported variable levels of transgene expression (Lay et al. 2004; Recillas-Targa et al. 2004; Emery 2011). This is mostly the consequence of the incompleteness of the regulatory region included in the transgene and the effect of the integration site. "
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