Hemocyanin contributes to embryonic adaptation to hypoxia in the migratory locust

Ambient hypoxia can pose a major threat to the survival of metazoan organisms, especially insect embryos. Hemocyanin exhibits dominant expression in insect embryos, but its specific roles in hypoxia adaptation remain unexplored. Soil-dwelling locust eggs may frequently experience hypoxia during development. A comprehensive analysis of physiological and biochemical characters of hemocyanin was conducted in the embryos of migratory locust Locusta migratoria. Our results demonstrated that the revolution process was the critical hypoxia-sensitive event during locust embryogenesis. Hemocyanin presented a prominent expression in the revolution stage and exhibited strong responses to hypoxia. The relative duration of revolution was correlated negatively with the expression of hemocyanin subunit 2 (HC2), suggesting that HC2 might be closely associated with hypoxia adaptation of locust embryos. Furthermore, a HC2 mutant locust strain was established using the CRISPR/Cas9 technology, and higher hypoxia sensitivity was found for HC2-deficient locust embryos. Knockdown of HC2 increased anaerobic metabolism and oxidative stress while reducing oxidative metabolism. Overall, these findings clearly demonstrated the pivotal roles of hemocyanin in hypoxia adaptation of insect embryos.