Simultaneous injection of NOS inhibitors with atropine interfered with the prevention of myopia (atropine + L-NIO, dRE: −15.6 ± 2.6 D, p < 0.0001, n = 12; dAL: 0.61 ± 0.2 mm, p = 0.002, n = 12; atropine + L-NMMA, dRE: −17.7 ± 2.1 D, p < 0.0001, n = 9; dAL: 0.56 ± 0.2 mm, p = 0.0321, n = 12). This evidence concerns the gene NOS1 and myopia.