Identification of tumor necrosis factor receptor 1 and its association with LPS-induced apoptosis in Onchidium reevesii

Abstract

The sea slug (Onchidium reevesii) inhabits intertidal environments and face environmental pressures and disease threats. The immune system of mollusks plays a crucial role in their defense against pathogens and maintaining homeostasis. The molecular mechanisms underlying its immune defense and response to pathogens remain largely unexplored. This study aimed to isolate and identify the cDNA sequence of tumor necrosis factor receptor 1 (TNFR1) in the sea slug. The obtained cDNA sequence was named OrTNFR1. Like other members of the TNF receptor superfamily, OrTNFR1 contains a signal peptide, TNF homologous domain, and death-related domain. qPCR results showed that OrTNFR1 was highly expressed in the hepatopancreas, blood, and intestines of sea slug. This indicates that OrTNFR1 plays a role in the immune-related tissues of sea slug and is consistent with the broad expression of TNFR1 in bivalves. To investigate the inducible expression pattern and downstream effects of OrTNFR1 in response to lipopolysaccharide (LPS) stimulation, we studied the functional role of the OrTNFR1 gene by stimulating with LPS in vivo and suppressing OrTNFR1 expression using siRNA technology. Our findings demonstrate that LPS stimulation rapidly up-regulates the expression of OrTNFR1 in the hepatopancreas, blood, and intestine of the sea slug. Moreover, si-TNFR1 effectively inhibits LPS-induced TNFR1 expression and its downstream genes. Additionally, we discovered that inhibiting OrTNFR1 reduces LPS-induced cell apoptosis and the expression of apoptosis-associated protein Caspase.TNFR1 is involved in the immune defense of the sea slug, and blocking TNFR1 effectively attenuates LPS-induced cell apoptosis through the TNFR1 pathway. This finding suggests that inhibiting TNFR1 in the sea slug may serve as a potential approach to reduce cell apoptosis. As a shell-less living in harsh intertidal environments, TNFR1 likely participates in its innate immunity. Therefore, studying the TNFR1 pathway contributes to understanding the innate immune signaling pathway in the sea slug, further elucidating host defense against pathogens and apoptotic functions.