Identification of phosphatidylserine receptor (PSR) gene can regulate zebrafish brain development through controlling on ROS stress and autophagy induction

Abstract

Zebrafish model system is important in genetic and developmental study with fully established online gene library available for gene function study short development period and transparent embryo make researchers fully access to all development stage with very low cost this make zebrafish a power model in gene study Phosphatidylserine receptor (PSR) locate on macrophage can identify apoptotic cell s which present phosphatidylserine that originally locate on inner membrane on cell surface Previous studies show that mouse without PSR have defect in brain eye and heart development PSR knock down in zebrafish embryo have identical result In our previous study PSR knock out zebrafish line was established by CRISPR/Cas9 system targeting in PSR gene on exon 3 Increased reactive oxygen species was detected in PSR knockout zebrafish embryo during gastrulation stage swelling heart and smaller brain at 24 hour post fertilization (hpf) In this study we investigate the gene profile in PSR knockout zebrafish embryo by CRISPR/Cas9 approach Then we use the NGS analysis at 8 hpf for further evaluation We found that mid-brain and hindbrain defected embryos have induced ROS-mediated stress signal and stress-related genes Then the novel genes downregulation of autophagy-related genes such as Beclin-1 ATG5 and ATG8 Finally the brain-defect embryos was rescued by extra mRNA of PSR by microinjection that ROS-mediated stress response and downregulation of autophagy-related genes is blocked Our results provide new insights in PSR-mediated engulfing signal for brain development

Date of Award	2021
Original language	English
Supervisor	Jiann-Ruey Hong (Supervisor)