Background: Three-thirds of people with radiologically isolated syndrome (RIS) develop multiple sclerosis (MS) within five years following their first brain magnetic resonance imaging (MRI). Subclinical applications of optical coherence tomography (OCT) include measuring the thickness of different retinal layers and monitoring the progression of visual pathway atrophy and neurodegeneration in relation to the progress of the entire brain. Objectives: Our OCT study was conducted in individuals with RIS to evaluate the thickness of the macular retinal nerve fiber layer (mRNFL) and the retinal ganglion cell layer (RGCL). Methods: In this study, 22 patients with RIS and 23 healthy individuals healthy control (HC) were enrolled. The control group and the RIS subjects underwent retinal imaging with OCT. Results: Total mRNFL thickness was 110.34 ± 13.71 μm in the RIS patients and 112.10 ± 11.23 μm in the HC group. Regional analysis of the mRNFL showed that the difference in thickness was more prominent in the superior quadrant. In regards to ganglion cell layer (GCL)++ thickness, the RIS and HCs population showed statistically significant differences in the nasal (P = 0.041), inferior (P = 0.040), and superior (P = 0.045) quadrants. The nasal (P = 0.041) quadrant showed the highest reduction in thickness compared to other regions of the GCL++. Meanwhile, no significant reduction was seen in GCL+ thickness (P-value > 0.05). When the thickness of the retinal layer of the right eye was compared to that of the left eye of the RIS group, no statistically significant differences were found (P-value > 0.05). Conclusions: Compared to the control group, the RIS group had a lower mean thickness of mRNFL and GCL++, indicating retinal neuroaxonal loss.