Cellular Localization of MAPK, NF-κB and Nrf2 Signaling Pathways-Related Proteins in Crayfish Hepatopancreas
Cellular Localization of MAPK, NF-κB and Nrf2 Signaling Pathways-Related Proteins in Crayfish Hepatopancreas
Keqiang Wei1*, Yue Wei2 and Changxia Song1
ABSTRACT
Crustaceans rely completely on their innate immune system comprised of various cells, molecules and signaling pathways to survive. Although signaling pathways such as MAPK, NF-κB and Nrf2 have been extensively studied in vertebrates, our current understanding of them in crustaceans is still in the infancy. Studies have found that three members, i.e. p38, RelA (p65) and Nrf2 are evolutionarily conserved, but the specific cells responsible for their synthesis and function have not been well characterized in crustaceans. In this work, we applied an immunohistochemical method to investigate their cellular localization and expression in hepatopancreas of red swamp crayfish (Procambarus clarkii), a commercially important aquaculture species in China. Using polyclonal antibodies, the strong staining of these proteins in hepatopancreatic cells was observed. The synchronous expression of p38 and its phosphorylated form (p-p38) was mainly present in B- and R- cells, and their MOD values were 0.0055±0.0038 and 0.0046±0.0027, respectively. As its downstream transcription factors, both RelA (p65) and Nrf2 were widely distributed in the cytoplasm of B- and R- cells, but RelA (p65) showed a relatively higher MOD level compared to Nrf2 (0.0097±0.002 versus 0.0039±0.004). All proteins except Nrf2 could be also obviously observed in sinusoids. The information above implied that B- and R- cells could be predominantly responsible for their function. This study will help us better understand immunity mechanisms of crustacean and promote disease control.
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