PURPOSE: This study assessed the autonomic nervous system in patients with central serous chorioretinopathy (CSC) by simultaneously measuring pupillary responses and heart rate variability (HRV). METHODS: We recruited 33 patients with CSC and 26 age- and sex-matched healthy controls. Using a pupillometry and acceleration plethysmography system, we measured the participants\' pupillary light reflex and HRV simultaneously, and compared the following parameters between the two groups: the pupil diameters, diameter changes, and time and frequency domain HRV indices (high frequency power: HF; low frequency power: LF; log LF/HF ratio). Additional data from the Profile of Mood States (POMS) and pupillary responses during mental tasks were also analyzed. RESULTS: The CSC group had a significantly lower constriction amplitude and a higher re-dilation ratio compared with the control group, indicating parasympathetic inhibition and sympathetic activation. For the HRV measures, the CSC group demonstrated significantly lower HF and higher LF and log LF/HF ratio, indicative of higher sympathetic activity. The CSC group also showed significantly larger pupil dilation during tasks of moderate difficulty, and higher negative/lower positive POMS mood scores. Further analyses also revealed that the baseline pupil diameter was significantly larger in patients with active as opposed to chronic CSC. CONCLUSIONS: Pupillary responses and HRV measures both revealed sympathetic activation and parasympathetic attenuation in patients with CSC. Larger pupil dilation during mental tasks in CSC could be a potential marker of psychophysiological stress.