Hprt1^{tm15(Ple111-EGFP)Ems}

The Ple111-EGFP transgene (pEMS1417) was designed with the 544 bp Ple111 minipromoter (HCRT-A; derived from a subsection of the promoter from the human hypocretin (orexin) neuropeptide precursor (HCRT)) upstream of a minimal F5 mutant-frt site, an enhanced green fluorescent protein (with mutated TAA->TTA stop), a nuclear localization signal, a second minimal frt site, an SV40 early polyA signal, and a human HPRT complementary sequence (containing exon1, intron1, exon2, and part of intron2). This construct was targeted as a single copy knockin to the Hprtb-m3 mutant locus on the X chromosome. The promoter/regulatory regions of the human hypocretin (orexin) neuropeptide precursor (HCRT) directs expression of an enhanced green fluorescent protein (EGFP) to a small cluster of EGFP-positive neurons is found in the hypothalamus. EGFP-immunoreactivity fills the cytoplasm of these cells, highlighting the cell body as well as extensive dendritic processes. This label is very specific to the hypothalamus and not seen in any other cell populations in the brain. Double label immunocytochemistry shows that a majority of these EGFP-positive hypothalamic cells co-express with the HCRT (orexin) neuropeptide (the gene from which this MiniPromoter was designed). (J:144244, J:164356)