Mori T et al. (2017), The constant threat from a non-native predator ...

XB-ART-54292

Biol Open 2017 Nov 15;611:1726-1733. doi: 10.1242/bio.029926.

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The constant threat from a non-native predator increases tail muscle and fast-start swimming performance in Xenopus tadpoles.

Mori T , Yanagisawa Y , Kitani Y , Yamamoto G , Goto-Inoue N , Kimura T , Kashiwagi K , Kashiwagi A .

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Predator-induced phenotypic plasticity is the ability of prey to adapt to their native predator. However, owing to environmental changes, encounters with unknown predators are inevitable. Therefore, study of prey and non-native predator interaction will reveal the primary stages of adaptive strategies in prey-predator interactions in the context of evolutionary processes. Here, Xenopus tadpoles exposed to a non-native predator, a larval salamander, showed a significant increase in body weight and tail length to body length ratio. The Tmax2 test indicated a significant enhancement of the tail muscle and decrease in the relative ventral fin height in tadpoles exposed to predation risk, leading to significantly higher average swimming speeds. The analysis of muscle-related metabolites revealed that sarcosine increased significantly in tadpoles exposed to non-native predators. Multiple linear regression analysis of the fast-start swimming pattern showed that the fast-start swimming speed was determined by the time required for a tadpole to bend its body away from the threat (C-start) and the angle at which it was bent. In conclusion, morphological changes in tadpoles were functionally adaptive and induced by survival behaviors of Xenopus tadpoles against non-native predators.

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Species referenced: Xenopus
Genes referenced: pigy

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	Fig. 1. Experimental design and changes in body length and weight in tadpoles exposed to predation. (A) Experimental design of the induced predation in Xenopus tadpoles. Xenopus tadpoles (50 tadpoles per aquarium) were assigned to five different treatment groups. The control group (Cont) was not exposed to the predator for 8 or 10 days of the experiment. The Exp 8 and Exp 10 groups were exposed to the predator for the full 8 and 10 days, respectively. (B) Body length in Xenopus tadpoles exposed to predation. (C) Morphology of Xenopus tadpoles. Tail length was determined as the distance from the anus to the tail top. (D,E) Body weight (D) and the ratio of tail length to body length (E) in tadpoles exposed to predation for 8 days. The number of tadpoles in Cont 8 and Exp 8 was 148 and 97, respectively. Statistical analysis was performed using Krishnaiah’s Embedded Image-test.
	Fig. 2. Analysis of muscle-related metabolites using CE-TOFMS. Ten different Xenopus tadpoles were randomly selected from the group of tadpoles exposed to predation for 8 days (Exp 8) and the control group (Cont 8). Then, the tadpoles were frozen in liquid nitrogen and metabolites extracted with methanol. A pool of methanol extracts from 10 tadpoles was treated as one sample and analyzed using CE-TOFMS.
	Fig. 3. Analysis of sarcosine using GC-MS. Tails from nine different Xenopus tadpoles were randomly selected from the group of tadpoles exposed to predation for 10 days (Exp 10) and the control group (Cont 10), and three tails were combined to prepare three samples for each experiment. Sarcosine was extracted and the content was expressed as picomol of sarcosine per gram of wet sample. Statistical analysis was performed using a two-sample t-test.
	Fig. 4. Measurements of tail height, tail muscle and height of the ventral fin. (A) Photographs of tadpoles in the control (Cont 10) and experimental group (Exp 10). Lines indicate tail height, and the lowercase letters a and b indicate height of the tail muscle and height of the ventral fin, respectively. Tail height (B), height of the tail muscle, and height of the ventral fin (C) were measured in 50 tadpoles from each group of tadpoles exposed to predation for 10 days (Exp 10) and the control (Cont 10). These values were obtained at the points of the greatest height of tadpole's tail from images of tadpoles using ImageJ software.
	Fig. 5. Swimming performance test. Ten tadpoles were randomly chosen from each experimental aquarium, and each tadpole was placed into an experimental electric chamber (A,B). After stabilizing a tadpole's behavior, an electric shock (12 V, 1.5 mA) was delivered to the tadpole for ∼300 ms (B). Swimming behavior of the tadpole was filmed with a high-speed camera at 240 frames/s (Movie 1), and the fast-start swimming speed was analyzed using tadpoles from the control (Cont 10) and the group of tadpoles exposed to predation for 10 days (Exp 10) (C). Statistical analysis was performed using a two-sample t-test after the Kolmogorov–Smirnov test.
	Fig. 6. Normal probability plot for the residual of the regression analysis of the fast-start swimming speed. All points of the normal plot for the residual were within a 95% confidence interval (CI, confidence interval; N, the number of residuals; AD, Anderson-Darling statistic; St Dev, standard deviation of residuals).

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