Immunohistochemical identification and quantitative analysis of cytoplasmic Cu/Zn superoxide dismutase in mouse organogenesis

Abstract

Cytoplasmic Cu/Zn superoxide dismutase (SOD1) is an antioxidant enzyme that converts superoxide to hydrogen peroxide in cells. Its spatial distribution matches that of superoxide production, allowing it to protect cells from oxidative stress. SOD1 deficiencies result in embryonic lethality and a wide range of pathologies in mice, but little is known about normal SOD1 protein expression in developing embryos. In this study, the expression pattern of SOD1 was investigated in post-implantation mouse embryos and extraembryonic tissues, including placenta, using Western blotting and immunohistochemical analyses. SOD1 was detected in embryos and extraembryonic tissues from embryonic day (ED) 8.5 to 18.5. The signal in embryos was observed at the lowest level on ED 9.5-11.5, and the highest level on ED 17.5-18.5, while levels remained constant in the surrounding extraembryonic tissues during all developmental stages examined. Immunohistochemical analysis of SOD1 expression on ED 13.5-18.5 revealed its ubiquitous distribution throughout developing organs. In particular, high levels of SOD1 expression were observed in the ependymal epithelium of the choroid plexus, ganglia, sensory cells of the olfactory and vestibulocochlear epithelia, blood cells and vessels, hepatocytes and hematopoietic cells of the liver, lymph nodes, osteogenic tissues, and skin. Thus, SOD1 is highly expressed at late stages of embryonic development in a cell- and tissue-specific manner, and can function as an important antioxidant enzyme during organogenesis in mouse embryos.

Differential Expression of Gastrointestinal Glutathione Peroxidase (GI-GPx) Gene during Mouse Organogenesis

“…However, the Sepp mRNA level in embryos is higher than in extraembryonic tissues on ED 14.5 and then appears at a similar level in both tissues until birth (Lee et al., 2008). Also, cytoplasmic Cu/Zn superoxide dismutase (SOD1), a primary antioxidant that catalyses the dismutation of superoxide radicals () to hydrogen peroxide, is increased at late stages of embryonic development, but maintains a constant level of expression in extraembryonic tissues during organogenesis (Yon et al., 2008a,b). These findings indicate that although various antioxidant enzymes are related to organogenesis, they have a different action mode in between the embryos and the extraembryonic tissues during embryogenesis.…”

Phospholipid hydroperoxide glutathione peroxidase is involved in the maintenance of male fertility under cryptorchidism in mice

“…To quantify protein concentrations in tissues, Western blotting was conducted as previously described [18]. Anti-PHGPx polyclonal antibody, anti-cytoplasmic GPx (GPx1) polyclonal antibody, and anti-GAPDH monoclonal antibody were purchased from Epitomics, Inc. (Burlingame, CA, USA), abcam (Cambridge, MA, USA), and Cell Signaling Technology (Beverly, MA, USA), respectively.…”

SOD1 Lysine 123 Acetylation in the Adult Central Nervous System

“…Previous immunohistochemical analysis identified high SOD1 immunoreactivity in olfactory sensory neurons (Yon et al, 2008), which are responsible for odor detection through their odorant receptors. Axons from olfactory sensory neurons project toward the main olfactory bulb, where they synapse with second-order neurons, the mitral and tufted cells, in specialized compartments called glomeruli.…”