Analysis of Drosophilia Sox Gene Expression in the Intestinal Stem Cell Lineage

Houda Boucekkine, John R. Nambu

Abstract


Vertebrate Sox2 has been shown to be required for mammalian stem cell pluripotency and generation of induced pluripotent stem cells (iPScs) (Takahashi et al., 2007). Drosophila Dichaete is homologous to Sox2 (Pevny et al., 1997; Phochanukul et al., 2010) and is related to many other genes within the Sox family, which is conserved in mammalian species. Thus, we wish to determine if Dichaete and related Sox genes perform a role in Drosophila stem cell maintenance, which may suggest that homologous Sox factors are important in mammalian stem cell maintenance. We propose to characterize the expression of Drosophila Dichaete and three closely related Group B Sox genes—Sox21a, Sox21b, and SoxNeuro (SoxN)—in the adult intestinal stem cell lineage (Ohlstein et al., 2006; Takashima et al., 2008). RT-PCR assays of intestinal tissue indicated that all four Group B Sox genes are expressed within the appropriate regions, and preliminary data from fluorescent reporter gene constructs revealed expression of both SoxN and Dichaete in subsets of intestinal cells. Immunostaining and/or in situ hybridization of dissected intestinal tissue will be used to confirm expression of Group B Sox genes within the intestine followed by a double labeling approach to characterize expression specifically within intestinal stem cells. If such expression is confirmed, this study will make a significant contribution towards unraveling the roles of Sox genes in stem cell biology and help to develop fruit fly intestinal stem cells as a genetic model for mammalian stem cells.


Keywords


Sox expression; stem cells; Drosophila model

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