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Abstract
Kisspeptins are neuropeptides encoded by the kiss1 gene, and little is known about them outside the vertebrate lineage. Two kisspeptin-type neuropeptides (KPs) have been discovered in Apostichopus japonicus (AjK1 and AjK2), an edible sea cucumber, and have been linked to reproductive and metabolic regulation. In this study, we evaluated how KPs affected locomotor behavior in one control group and two treatment groups (AjK1 and AjK2). We discovered that AjK1 had a significant dose effect, primarily by shortening the stride length and duration of movement to reduce the sea cucumber movement distance, whereas AjK2 had little inhibitory effect at the same dose. The levels of phosphatidylethanolamine (PE), phosphatidylcholine (PC), uridine, glycine, and L-serine in the longitudinal muscle of A. japonicus treated with AjK1 differed significantly from those of the control, which may explain the observed changes in locomotor behavior. Treatment with AjK2 induced changes in aspartate levels. Our results imply that AjK1 is more likely than AjK2 to have a role in the regulation of A. japonicus locomotion.
41876157 National Natural Science Foundation of China, COMS2019Q15 the Key Deployment Project of Centre for Ocean Mega-Research of Science, Chinese Academy of Sciences
Figure 1. The total distance moved (a) and cumulative moving duration (b) before and after injection for each group. The data in the figure are total measured data for 12 h. Each error bar represents the standard error. S is the seawater control group, K1 and K2 are the neuropeptide treatment groups, and H/M/L represent high, medium, and low concentrations. *, significant difference within the group (p < 0.05).
Figure 2. Trend of locomotor behavior of A. japonicus within 12 h after injection compared with before injection: (a) distance; (b) moving duration; (c) velocity; (d) step number; (e) stride length; (f) step frequency. Behavioral data were measured hourly, and the values in the figure are the differences (ΔY) at the same moment in the two adjacent days before and after injection. The error bars in the figure represent the standard error (n = 8 for each group). S is the seawater control group, K1 and K2 are the neuropeptide treatment groups, and H/M/L represent high, medium, and low concentrations.
Figure 3. Significance of differences between different neuropeptide groups changed over time: (a) distance; (b) moving duration; (c) velocity; (d) step number; (e) stride length; (f) step frequency. (a–c) show the difference between different dose levels (0/L/M/H) of K1 (or K2), which were compared by one-way ANOVA. (d–f) The difference between the two high concentration treatment groups (K1H and K2H) and the seawater control group, which were compared using t-tests (two-tailed). The significance level was set at 0.05.
Figure 4. PLS-DA scores plot of longitudinal muscle metabolite profiling. (a) CM (in blue) vs. K1M (in red) in positive ion mode, Q2
= 0.546; (b) CM (in blue) vs. K2M (in red) in positive ion mode, Q2
= 0.659; (c) K1M (in blue) vs. K2M (in red) in positive ion mode, Q2
< 0 (unreliable); (d) CM (in blue) vs. K2M (in red) in negative ion mode, Q2
= 0.717; (e) CM (in blue) vs. K2M (in red) in negative ion mode, Q2
= 0.895; (f) K1M (in blue) vs. K2M (in red) in negative ion mode, Q2
= 0.713.
Figure 5. Venn diagram of all differential metabolites from the experimental groups (K1M and K2M) both in positive and negative ion mode. Circles in orange indicate the AjK1 treatment group; circles in blue indicate the AjK2 treatment group; solid lines mean that metabolites are up-regulated; dashed lines mean that they are down-regulated.
Figure 6. Enrichment analysis of differential metabolic pathways. K1M is shown in orange and K2M is shown in blue. The vertical line mean p = 0.05.
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