The purpose of this study is to ascertain whether inherent differences exist in the retinal function of two macaque species that are commonly used in experimental glaucoma investigations. We quantified and compared multifocal electroretinograms (mfERGs) from ocularly normal cynomolgus (n = 6) and rhesus (n = 8) monkeys. The stimulus array consisted of 103 equal-sized hexagonal elements, which subtended ±44° about the central visual axis. Mean luminance of the display was 100 cd/m2. The first-order kernel (K1) and second-order (first slice) kernel (K2) of the mfERG were averaged into 4 rings radiating from the foveal element, and represented the central 56° of visual field. Fifteen and 30-element segments were used for K1 and K2 quadrant and hemiretinal response determinations, respectively. Response measures for the rings, quadrants, and hemiretinae included K1 amplitude and implicit time, and K1 and K2 oscillatory potentials (OPs) and response amplitude root mean square (RMS). Species, gender, and retinotopic differences were assessed with repeated measures analysis of variance (split plot design). K1 amplitudes of the N1 waves, K1 and K2 OPs and K2 amplitude RMS for the ring, quadrant, and hemiretinal mfERG waveforms were larger in rhesus than in cynomolgus monkeys. Rhesus males (as compared to rhesus females) and cynomolgus females (as compared to cynomolgus males) exhibited larger amplitudes and less delayed implicit times in the central retina. These results demonstrate that species-specific differences in retinal function are evident in cynomolgus and rhesus monkeys. There also were gender-associated differences that varied across species. Thus, investigators should exercise caution when data from species or gender are combined.