Schematic diagram of NPY-Cre-GFP knock-in strategy and confirmation of Y2 receptor deletion.

Abstract

<p>(A) Conditional NPY neuron-specific Y2 receptor knockout mice (NPYCre/+;Y2^lox/lox mice) were generated using a knock-in strategy. The inducible NPY neuron-specific expression of the Cre gene is achieved by fusion of the initiation codon of the NPY gene with that of the tetR gene, enabling the endogenous NPY promoter to drive the expression of this regulatory protein upon addition of doxycycline (Dox). (B) Schematic drawing of position of oligonucleotide primers (Oligo A & B) used for PCR verification of Y2 receptor knockout. (C) Using genomic DNA extracted from the hypothalamus of NPYCre/+;Y2^lox/lox mice, Oligos A and B produced a 250 base pair (bp) product from a Dox-injected mouse, demonstrating deletion of the Y2 receptor gene, but no product was produced using DNA from a saline-injected control mouse. (D) Y2 receptor mRNA extracted from the hypothalamus of male NPYCre/+;Y2^−/− mice was significantly reduced compared to that of saline-injected mice.</p