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. 2020 Dec 3;10(1):21141.
doi: 10.1038/s41598-020-78316-z.

Autophagy: a necessary defense against extreme cadmium intoxication in a multigenerational 2D experiment

Agnieszka Babczyńska  1 , Agnieszka Nowak  2 , Alina Kafel  2 , Bartosz Łozowski  2 , Magdalena Rost-Roszkowska  2 , Monika Tarnawska  2 , Maria Augustyniak  2 , Marta Sawadro  2 , Agnieszka Molenda  2
Affiliations

Autophagy: a necessary defense against extreme cadmium intoxication in a multigenerational 2D experiment

Agnieszka Babczyńska et al. Sci Rep. .

Abstract

Autophagy is a natural process that aims to eliminate malfunctioning cell parts, organelles or molecules under physiological conditions. It is also induced in response to infection, starvation or oxidative stress to provide energy in case of an energy deficit. The aim of this 2-dimensional study was to test if, and if so, how, this process depends on the concentration of cadmium in food (with Cd concentrations from 0 to 352 μg of Cd per g of food (dry weight)-D1 dimension) and the history of selection pressure (160 vs 20 generations of exposure to Cd-D2 dimension). For the study, the 5th instar larvae of a unique strain of the moth Spodoptera exigua that was selected for cadmium tolerance for 160 generations (44 μg of Cd per g of food (dry weight)), as well as 20-generation (11, 22 and 44 μg of Cd per g of food (dry weight)) and control strains, were used. Autophagy intensity was measured by means of flow cytometry and compared with life history parameters: survivability and duration of the 3rd larval stage. The highest values of autophagy markers were found in the groups exposed to the highest Cd concentration and corresponded (with a significant correlation coefficient) to an increased development duration or decreased survivorship in the respective groups. In conclusion, autophagy is probably initiated only if any other defense mechanisms, e.g., antioxidative mechanisms, are not efficient. Moreover, in individuals from pre-exposed populations, the intensity of autophagy is lower.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
AIR (A. induction ratio) and MAI (Mean a. intensity) in the hemolymph and midgut of the 5th larval stage of the moth S. exigua from the 2D experiment. Means ± mean and max values. Different letters (A, B, C) denote statistically significant differences between strains (D1); ANOVA, Tukey’s test for unequal sample size, p ≤ 0.05.
Figure 2
Figure 2
AIR in the hemolymph of the 5th larval stage of the moth S. exigua from the 2D experiment. D1, D2—dimensions of the experiment, according to Fig. 10. Values of the AIR are expressed according to a color scale, where dark green is the lowest and dark red is the highest. Different letters (a, b, c) denote statistically significant differences between experimental groups (D2); ANOVA, Tukey’s test for unequal sample size, p ≤ 0.05.
Figure 3
Figure 3
MAI in the hemolymph of the 5th larval stage of the moth S. exigua from the 2D experiment. D1, D2—dimensions of the experiment, according to Fig. 10. Values of the MAI are expressed according to a color scale, where dark green is the lowest and dark red is the highest. different letters (a, b) denote statistically significant differences between experimental groups (D2); ANOVA, Tukey’s test for unequal sample size, p ≤ 0.05.
Figure 4
Figure 4
Euclidean distance clustering tree describing the relationships among autophagy parameter (AIR and MAI) values of hemocytes of the 5th larval stage of the moth S. exigua from the 2D experiment. Symbols: strain-experimental group.
Figure 5
Figure 5
AIR in the midgut of the 5th larval stage of the moth S. exigua from the 2D experiment. D1, D2—dimensions of the experiment, according to Fig. 10. Values of the AIR are expressed according to a color scale, where dark green is the lowest and dark red is the highest. Different letters (a, b) denote statistically significant differences between experimental groups (D2); ANOVA, Tukey’s test for unequal sample size, p ≤ 0.05.
Figure 6
Figure 6
MAI in the midgut of the 5th larval stage of the moth S. exigua from the 2D experiment. D1, D2—dimensions of the experiment, according to Fig. 10. Values of the MAI are expressed according to a color scale, where dark green is the lowest and dark red is the highest. Different letters (a, b) denote statistically significant differences between experimental groups (D2); ANOVA, Tukey’s test for unequal sample size, p ≤ 0.05.
Figure 7
Figure 7
Euclidean distance clustering tree describing the relationships among autophagy parameter (AIR and MAI) values of midgut cells of the 5th larval stage of the moth S. exigua from the 2D experiment. Symbols: strain-experimental group.
Figure 8
Figure 8
Duration of the 3rd larval instar of the moth S. exigua from the 2D experiment. D1, D2—dimensions of the experiment, according to Fig. 10. Values are expressed according to a color scale, where dark green is the lowest and dark red is the highest. Capital letters (A, B, C) denote statistically significant differences between strains (D1), and small letters (a, b, c) denote statistically significant differences between experimental groups (D2); ANOVA, Tukey’s test for unequal sample size, p ≤ 0.05.
Figure 9
Figure 9
Cumulative mortality of 3rd and 4th larval instars of the moth S. exigua from the 2D experiment. D1, D2—dimensions of the experiment, according to Fig. 10. Values are expressed as the % of living individuals and according to a color scale, where dark green is the lowest and dark red is the highest mortality.
Figure 10
Figure 10
Scheme of the 2D experiment. Asterisks connect the timepoint of the experiment and the experimental groups. D1—strains, D2—experimental groups.
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