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Glutathione peroxidase 3 (GPx3) belongs to the glutathione peroxidase family of selenoproteins and is a key antioxidant enzyme in multicellular organisms against oxidative damage. Downregulation of GPx3 affects tumor progression and metastasis and is associated with liver and heart disease. However, the physiological significance of GPx3 in vertebrate embryonic development remains poorly understood. The current study aimed to investigate the functional roles of  gpx3  during embryogenesis. To this end, we determined  gpx3 ’s spatiotemporal expression using  Xenopus laevis  as a model organism. Using reverse transcription polymerase chain reaction (RT-PCR), we demonstrated the zygotic nature of this gene. Interestingly, the expression of  gpx3  enhanced during the tailbud stage of development, and whole mount in situ hybridization (WISH) analysis revealed  gpx3  localization in prospective tail region of developing embryo.  gpx3  knockdown using antisense morpholino oligonucleotides (MOs) resulted in short post-anal tails, and these malformed tails were significantly rescued by glutathione peroxidase mimic ebselen. The gene expression analysis indicated that  gpx3  knockdown significantly altered the expression of genes associated with Wnt, Notch, and bone morphogenetic protein (BMP) signaling pathways involved in tailbud development. Moreover, RNA sequencing identified that  gpx3  plays a role in regulation of cell death in the developing embryo. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and phospho-histone 3 (PH3) staining confirmed the association of  gpx3  knockdown with increased cell death and decreased cell proliferation in tail region of developing embryos, establishing the involvement of  gpx3  in tailbud development by regulating the cell death. Furthermore, these findings are inter-related with increased reactive oxygen species (ROS) levels in  gpx3  knockdown embryos, as measured by using a redox-sensitive fluorescent probe HyPer. Taken together, our results suggest that  gpx3  plays a critical role in posterior embryonic development by regulating cell death and proliferation during vertebrate embryogenesis.

Xenopus gpx3 Mediates Posterior Development by Regulating Cell Death during Embryogenesis