The removal of the internal limiting membrane (ILM) for traction maculopathy has recently been advocated. However, it is generally believed that the ILM plays an important role in retinal function, because it is the basal lamina of the Müller cells that are involved in the generation of the electroretinogram (ERG) b-wave. To date, there has been no objective assessment of retinal function on removing the ILM. In this study, the changes of each component of the focal macular electroretinograms (FMERGs) were investigated in eyes before and after the ILM was removed in the macular area during surgery for idiopathic macular holes (IMHs). FMERGs were elicited by a 15 degrees stimulus centered on the fovea and monitored by an infrared fundus camera. FMERGs were recorded from 49 eyes of 48 patients with IMHs before and 6 weeks after anatomically successful macular hole surgery. Whether an eye had or did not have the ILM removed was randomly determined. The ILM was removed in 30 eyes (ILM-off group) and was not removed in 19 eyes (ILM-on group). Six months after surgery, the same examination was performed in 27 eyes of the ILM-off group and in 15 eyes of the ILM-on group. The amplitudes and implicit times of the a- and b-waves and the mean amplitudes and implicit times of the first three oscillatory potentials (OP1 to OP3) were compared before and after surgery within and between the groups. Visual acuity increased significantly after surgery in both groups. In the ILM-on group, the amplitude of the a- and b-waves and the OPs increased significantly 6 months after surgery (P: = 0.0093, P: = 0.0019, P: = 0.0024, respectively, paired t-test). In the ILM-off group, the a-wave amplitude and mean OP amplitudes were significantly larger 6 months after surgery (P: = 0.0077, P: = 0.0030, respectively, paired t-test). The b-wave amplitude, however, did not change significantly. The percentage increase in the b-wave amplitude 6 months after surgery was significantly higher in the ILM-on group (44.0%) than in the ILM-off group (15.0%; P: = 0.037, t-test). The removal of the ILM had no adverse effect on visual acuity. However, the selective delay of recovery of the FMERG b-wave 6 months after surgery suggests an alteration of retinal physiology in the macular region.