Abstract: Early-life stress (ELS) refers to a period of environmental/social deprivation, physical, sexual or emotional abuse, neglect, severe and/or chronic trauma in the prenatal/early postnatal stage, which is closely related to many adverse psychiatric disorders later in life, such as depressive disorder, substance abuse, dysthymia, panic, anxiety, and suicidal behavior in adulthood (Waters and Gould, 2022). In addition to mood-related symptoms, a strong association between ELS and a wide range of adverse health outcomes in adulthood has been demonstrated, with increased risk of metabolic, musculoskeletal, neurological, respiratory, cardiovascular, and gastrointestinal symptoms (Wegman and Stetler, 2009). Therefore, ELS sequelae represent a serious public health concern. During childhood, social ties (mainly mother-infant relationship) play a key role in brain development and behavior in adulthood, and neonatal maternal separation (MS) has been shown to dramatically affect brain development, and increase the risk for several diseases (Cao et al., 2020). Since working with humans subjected to ELS is limited by the difficulty of finding a representative sample that experiences early adversities at the same stage, intensity, and duration, experimental approaches in animals are necessary to gain a deeper understanding of ELS-induced long-term consequences (Waters and Gould, 2022). In rodents, as in other mammals, the early postnatal environment is strongly determined by mother-pup interactions. In this regard, a mouse model of MS is widely recognized as a suitable model to mimic early adverse experiences, increasing depressive-like and/or anxiety-like behaviors in adulthood (Nishi, 2020), as demonstrated by behavioral tests, such as the sucrose preference test, the forced swim test, the open field test, the elevated plus-maze test, and the tail suspension tests (Nishi, 2020; Waters and Gould, 2022), some of which depend on the integrity of the visual system. In adult female Wistar rats, MS has been shown to induce a significant decrease in the thickness of the outer nuclear layer of the retina, as well as an increased retinal microglial reactivity and glial fibrillary acidic protein (a gliotic marker)-immunoreactivity (Grigoruta et al., 2020). However, despite that numerous effects of ELS on the central nervous system have been reported, its effects on the retina and visual system of adult mice remain unknown.