Crustacean hyperglycemic hormone (CHH) affects hemocyte intracellular signaling pathways to regulate exocytosis and immune response in white shrimp Litopenaeus vannamei

Recombinant Litopenaeus vannamei CHH (rLvCHH) was obtained from a bacterial expression system and the intracellular signaling pathways involved in exocytosis and immune response after rLvCHH injection (0.2 and 2 μg/shrimp) was investigated in this study. The results showed that CHH contents increased 51.4%–110.2% (0.2 μg/shrimp) and 65.0%–211.3% (2 μg/shrimp) of the control level. And the contents of three biogenic amines in hemolymph presented a similar variation pattern after rLvCHH injection, but reached the highest level at different time points. Furthermore, the mRNA expression levels of membrane-bound guanylyl cyclase (mGC) (1.20–1.93 fold) and biogenic amine receptors, including type 2 dopamine receptor (DA2R) (0.72-0.89 fold), α2 adrenergic receptor (α2-AR) (0.72-0.91 fold) and 5-HT7 receptor (5-HT7R) (1.37–3.49 fold) in hemocytes were changed consistently with their ligands. In addition, the second messenger and protein kinases shared a similar trend and reached the maximum at the same time respectively. The expression levels of nuclear transcription factor (cAMP response element-binding protein, CREB) and exocytosis-related proteins transcripts were basically overexpressed after rLvCHH stimulation, which reached the peaks at 1 h or 3 h. Eventually, the phenoloxidase (PO) activity (37.4%–158.5%) and antibacterial activity (31.8%–122.3%) in hemolymph were dramatically enhanced within 6 h, while the proPO activity in hemocytes significantly decreased (11.2%–62.6%). Collectively, these results indicate that shrimps L. vannamei could carry out a simple but ‘smart’ NEI regulation by releasing different neuroendocrine factors at different stages after rLvCHH stimulation, which could couple with their receptors and trigger the downstream signaling pathways during the immune responses in hemocytes.