Silencing two main isoforms of crustacean hyperglycemic hormone (CHH) induces compensatory expression of two CHH-like transcripts in the red swamp crayfish Procambarus clarkii

Abstract

RNA interference has frequently been applied to modulate gene function in organisms. With the aim of creating new autocidal methods based on neuro-endocrine disruptors for invasive populations of Procambarus clarkii, we silenced the Crustacean Hyperglycemic Hormone (CHH) by injecting the corresponding dsRNA. CHH is a pleiotropic hormone that primarily regulates the mobilization of energy reserves and plays a pivotal role in stress responses. Here, we describe two experiments aimed at testing whether CHH silencing significantly alters important physiological aspects. The first experiment investigates the effects of CHH silencing at the glycemic and transcriptomic level in the eyestalk. The second experiment explores the long-term effects of CHH silencing and the effects on mortality and moulting rates. Osmotic deficits and mortality were recorded in specimens injected with CHH dsRNA, whilst controls were injected with GFP dsRNA. After 20 days, despite still silenced for CHH, individuals that survived recovered a strong hyperglycemic response after serotonin injection due to the compensatory effect of two peptides belonging to the crustacean neurohormone CHH protein family.