Deficient working memory contributes to cognitive impairment in DS. To assess working memory in Ts65Dn:Kcnj6++− mice, the rate of spontaneous alternations in Y-maze was measured. The alternation rate was significantly lower in Ts65Dn:Kcnj6+++ vs. 2N:Kcnj6++ group (F1,36 = 3.06; p = 0.002) reflecting impaired working memory (Fig. 4A, red vs. blue bar). Reducing Kcnj6 gene dose had no effect on the alternation rate (F1,28 = 0.23; p = 0.41 for Ts65Dn:Kcnj6+++ vs. Ts65Dn:Kcnj6++−) thus demonstrating no effect in this test of working memory.