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. 2018 Mar 22:9:269.
doi: 10.3389/fphys.2018.00269. eCollection 2018.

Melatonin Promotes Cheliped Regeneration, Digestive Enzyme Function, and Immunity Following Autotomy in the Chinese Mitten Crab, Eriocheir sinensis

Cong Zhang  1   2   3 , Xiao-Zhen Yang  1   2   3 , Min-Jie Xu  1   2   3 , Gen-Yong Huang  1   2   3 , Qian Zhang  1   2   3 , Yong-Xu Cheng  1   2   3 , Long He  1   2   3 , Hong-Yu Ren  1   2   3
Affiliations

Melatonin Promotes Cheliped Regeneration, Digestive Enzyme Function, and Immunity Following Autotomy in the Chinese Mitten Crab, Eriocheir sinensis

Cong Zhang et al. Front Physiol. .

Abstract

In the pond culture of juvenile Eriocheir sinensis, a high limb-impairment rate seriously affects the culture success. Therefore, it is particularly important to artificially promote limb regeneration. This study evaluated the effects of melatonin on cheliped regeneration, digestive ability, and immunity, as well as its relationship with the eyestalk. It was found that the injection of melatonin significantly increased the limb regeneration rate compared with the saline group (P < 0.05). The qRT-PCR results of growth-related genes showed that the level of EcR-mRNA (ecdysteroid receptor) and Chi-mRNA (chitinase) expression was significantly increased following the melatonin injection, while the expression of MIH-mRNA (molt-inhibiting hormone) was significantly decreased (P < 0.05). Melatonin significantly increased lipase activity (P < 0.05). We observed that the survival rates of limb-impaired and unilateral eyestalk-ablated crabs were substantially improved following melatonin treatment, whereas the survival of the unilateral eyestalk-ablated crabs was significantly decreased compared with the control group (P < 0.05). Furthermore, the results of serum immune and antioxidant capacity revealed that melatonin significantly increased the total hemocyte counts (THC), hemocyanin content, total antioxidant capacity (T-AOC), acid phosphatase (ACP), and glutathione peroxidase activity (GSH-Px), whereas the immune-related parameters were significantly decreased in eyestalk-ablated crabs (P < 0.05). Therefore, these findings indicate that melatonin exerts a protective effect on organism injury, which could promote limb regeneration by up-regulating the expression of growth-related genes, improve digestive enzyme activity, and strengthen the immune response, particularly antioxidant capacity.

Keywords: Eriocheir sinensis; digestive enzyme; eyestalk; immunity; melatonin; regeneration.

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Figures

Figure 1
Figure 1
The cheliped bud length of E. sinensis over 12 days following the different treatments. The values are expressed as the means ± SD (n = 10). Different letters placed above column represent significant differences between treatments (P < 0.05).
Figure 2
Figure 2
The level of EcR, MIH, and Chi gene expression normalized to β-actin in the hepatopancreas, muscle, epidermal, midgut, and hindgut of E. sinensis for each of the different treatments. (A) EcR, ecdysteroid receptor; (B) MIH, molt-inhibiting hormone; and (C) Chi, chitinase. The values are expressed as the means ± SD (n = 5). Different letters placed above column represent significant differences between treatments (P < 0.05).
Figure 3
Figure 3
The survival rate of E. sinensis for all groups at the completion of the experiment. The values are expressed as the means ± SD (n = 3). Different letters placed above column represent significant differences between treatments (P < 0.05).
Figure 4
Figure 4
The level of THC in E. sinensis according to each treatment. The values are expressed as the means ± SD (n = 10). Different letters placed above column represent significant differences between treatments (P < 0.05).
Figure 5
Figure 5
The effect of unilateral eyestalk ablation and melatonin injection on the immune and antioxidant capacity in E. sinensis. (A) Hemocyanin content; (B) acid phosphatase (ACP) activity; (C) alkaline phosphatase (ALP) activity; (D) total antioxidant capacity (T-AOC); (E) superoxide dismutase (SOD) activity; and (F) glutathione peroxidase (GSH-PX) activity. The values are expressed as the means ± SD (n = 10). Different letters placed above column represent significant differences between treatments (P < 0.05).
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