The Potential Roles of the Apoptosis-Related Protein PDRG1 in Diapause Embryo Restarting of Artemia sinica.

Wan Zhang,Lin Hou,Na Li,Hong Zhang,Feng Yao,Linlin Sui,Xiangyang Zou

doi:10.3390/ijms19010126

Abstract

High salinity and low temperatures can induce Artemia sinica to enter the diapause stage during embryonic development. Diapause embryos stop at the gastrula stage, allowing them to resist apoptosis and regulate cell cycle activity to guarantee normal development after diapause termination. P53 and DNA damage-regulated gene 1 (pdrg1) is involved in cellular physiological activities, such as apoptosis, DNA damage repair, cell cycle regulation, and promotion of programmed cell death. However, the role of pdrg1 in diapause and diapause termination in A. sinica remains unknown. Here, the full-length A. sinica pdrg1 cDNA (As-pdrg1) was obtained and found to contain 1119 nucleotides, including a 228 bp open reading frame (ORF), a 233 bp 5′-untranslated region (UTR), and a 658-bp 3′-UTR, which encodes a 75 amino acid protein. In situ hybridization showed no tissue specific expression of As-pdrg1. Quantitative real-time PCR and western blotting analyses of As-pdrg1 gene and protein expression showed high levels at 15–20 h of embryo development and a subsequent downward trend. Low temperatures upregulated As-pdrg1 expression. RNA interference for the pdrg1 gene in Artemia embryos caused significant developmental hysteresis. Thus, PDRG1 plays an important role in diapause termination and cell cycle regulation in early embryonic development of A. sinica.

Highlights

Artemia sinica is a small aquatic crustacean living in an environment with a wide variation in temperature and high salt concentration [1,2]
We determined that the increased mRNA and protein expression levels of As-P53 and DNA damage control gene 1 (PDRG1) could promote apoptosis to remove unwanted, damaged, and dangerous cells during development and to maintain homeostasis [19]. These results suggested that As-PDRG1 plays an important role in diapause termination and early embryonic development of A. sinica
Since the pdrg1 has functions such as DNA damage repair, we examined the expression of As-pdrg1 under low temperature

Summary

Introduction

Artemia sinica is a small aquatic crustacean living in an environment with a wide variation in temperature and high salt concentration [1,2]. During embryonic development of A. sinica, high salinity, low temperature, light cycle changes, and lack of food can induce A. sinica to enter the embryo diapause stage. In process of embryo diapause, dormant cysts of A. sinica show strong resistance to long-term drought, cold, hypoxia, and other severe environmental factors. There has been wide interest in understanding the causes and molecular mechanisms of diapause and diapause termination of A. sinica embryo development. Our previous studies suggested that Myeloid differentiation factor 88 (As-MYD88) and Late embryogenesis abundant proteins(As-LEA) proteins might play vital roles in post-diapause embryonic development and stress tolerance in A. sinica [10,11]

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