A Preliminary Study Of The Effect Of Ferroptosis On Head Regeneration Of Planarians

Background: Regeneration is a complex biological process that requires the correct response of the body to injury and tissue reorganization,which requires precise regulation of processes such as cell proliferation,cell death.Cell death is very important for the normal development of the body,the maintenance of homeostasis and the prevention of diseases caused by excessive proliferation.Ferroptosis is a non-apoptotic form of programmed cell death resulting from intracellular lipid peroxides accumulation due to dysregulation of the glutathione-dependent antioxidant defense system.However,the role of ferroptosis in regeneration is still poorly understood.Methods and results: In this experiment,Dugesia japonica,which has a strong regeneration ability,was selected as the research object to explore the effect of ferroptosis on the regeneration of planaria.First,the expression patterns of ferroptosis marker genes in intact worms were analyzed by whole-mount in situ hybridization and single cell transcriptome database.It was found that ferroptosis related genes were mainly expressed in neoblasts,nerve cells,intestinal cells and epidermal cells of planarian.Subsequently,the effect of treatment of the ferroptosis inducer Erastin was on planarian head regeneration was evaluated by phenotypic observation,H&E staining,transmission electron microscope,immunostaining,ROS staining and real-time quantification PCR experiments.The results showed that 3μM Erastin treatment significantly impaired regeneration of planarian head ganglia and lateral nerve branches,reduced mitochondria number and affected mitochondria morphology,upregulated the ROS level,and disturbed the expression patterns of ferroptosisrelated genes.Finally,the results of H3 P immunofluorescence staining showed that cell proliferation at 6hpa,1dpa and 3dpa was significantly inhibited after treatment of Erastin.The results of TUNEL staining showed that Erastin treatment gave rise to increased cell death.Conclusion: This study suggests ferroptosis is involved in the process of planarian head regeneration.Ferroptosis induced by Erastin significantly inhibit planarian neoblasts proliferation,and it can also induce large-scale cells death in the planarian,leading to failure of head regeneration of planarian.This experiment explored the relationship between ferroptosis and planarian regeneration,and provided a new research foundation for further understanding of the internal mechanism of planarian regeneration.