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. 2023 Mar 30;11(4):327.
doi: 10.3390/toxics11040327.

Acute Exposure to Arsenic Affects Pupal Development and Neurological Functions in Drosophila melanogaster

Anushree  1 , Md Zeeshan Ali  1 , Anwar L Bilgrami  2 , Jawaid Ahsan  1
Affiliations

Acute Exposure to Arsenic Affects Pupal Development and Neurological Functions in Drosophila melanogaster

Anushree et al. Toxics. .

Abstract

Millions of people in developing countries are affected by arsenic (As) toxicity and its prevalence. Arsenic's detrimental effects on humans have been amplified by an unacceptable level of exposure to food and drinking water, the ongoing rise in industrial usage, and several other occupational conditions. Due to increased cellular absorption and the ability to cross the blood-brain barrier (BBB), inorganic arsenic (iAs) is extremely hazardous to living organisms in its trivalent form. Arsenic toxicity damages an organism's tissues and organs, resulting in skin cancer, circulatory system abnormalities, and central nervous system disorders. However, a competent model system is required to investigate the acute effects of arsenic on the brain, cognition ability, and to assess any behavioral impairment. Hence, Drosophila, with its short generation time, genomic similarities with humans, and its availability for robust behavioral paradigms, may be considered an ideal model for studying arsenic toxicity. The present study helps to understand the toxic effects of acute arsenic treatment on the behavior, cognition, and development of Drosophila in a time-dependent manner. We found that the exposure of fruit flies to arsenic significantly affected their locomotor abilities, pupae size, cognitive functions, and neurobehavioral impairment. Hence, providing a better understanding of how arsenic toxicity affects the brain leading to acute behavioral disorders and neurological alterations, this study will lead to a better understanding of the mechanisms.

Keywords: Drosophila melanogaster; LC50; arsenic; behavior; learning; toxicity.

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Conflict of interest statement

The authors have no relevant financial or non-financial interest to disclose.

Figures

Figure 1
Figure 1
An illustration of an experimental two-choice olfactory assay designed for adult Drosophila flies. The two traps attached to the Petri plate do not allow flies to change their choice, once they enter the glass vial. One glass vial contained 300 µL of 0.2% Triton X-100 diluted in water (solvent), while the other contained 300 µL of ethyl acetate odor of 10−2 dilution in 0.2% Triton X-100.
Figure 2
Figure 2
An illustration of an experimental T-maze testing assay designed for adult Drosophila flies. The two arms of the T-tube, the escape arm, and odorant arm are attached to the traps that do not allow the flies to change their choice, once they enter the trap. Parafilm helps in avoiding flies from being trapped in between the space and tight fitting of the trap with the T-tube. One trap contained 20 µL of mineral oil (solvent), while the other contained 20 µL of ethyl acetate odor of 10−5 dilution in mineral oil.
Figure 3
Figure 3
(A) The graph shows sodium arsenite (III) treated adult Drosophila melanogaster survivability within 24 h. Bars represent the means ± S.D. The significant mean difference (p < 0.05) between treated and untreated flies was analyzed by a one-way analysis of variance (ANOVA) (*** p < 0.0001). (B) The lethal concentration of sodium arsenite for flies within 24 h using the probit linear regression statistic is shown. Slope intercepts at 4.02 on the y-axis, log concentration is 0.29 mM, and the linear equation is y = 3.35x + 4.02 with R squared value as 0.89. The LC50 obtained is 1.96 mM.
Figure 3
Figure 3
(A) The graph shows sodium arsenite (III) treated adult Drosophila melanogaster survivability within 24 h. Bars represent the means ± S.D. The significant mean difference (p < 0.05) between treated and untreated flies was analyzed by a one-way analysis of variance (ANOVA) (*** p < 0.0001). (B) The lethal concentration of sodium arsenite for flies within 24 h using the probit linear regression statistic is shown. Slope intercepts at 4.02 on the y-axis, log concentration is 0.29 mM, and the linear equation is y = 3.35x + 4.02 with R squared value as 0.89. The LC50 obtained is 1.96 mM.
Figure 4
Figure 4
(A) The length of pupae that developed in 1 mM arsenite media and control media were measured and are represented. A decrease in the length (mm) of treated pupae compared to the control is apparent from the graph. The statistical significance (p < 0.05) was analyzed by the Wilcoxon signed rank test with *** p < 0.0001 and 95 per cent confidence interval. (B) The width of pupae that developed in 1 mM arsenite media and control media were measured and are represented. A decrease in the width (mm) of treated pupae compared to the control is apparent from the graph. The statistical significance (p < 0.05) was analyzed by the Wilcoxon signed rank test with *** p < 0.0001 and 95 per cent confidence interval.
Figure 5
Figure 5
The box plot represents the negative geotaxis ability of adult flies treated at different concentrations of arsenite compared to the control flies. The distance is measured in millimeters (mm). The upper bar displays the maximum value from the dataset, while the lower bar displays the minimum value from the dataset. The significant mean difference (p < 0.05) between the untreated and treated climbing ability was analyzed by a one-way analysis of variance (ANOVA) (** p < 0.01; *** p < 0.001; R square = 0.712).
Figure 6
Figure 6
The graph represents an olfactory response index of control, 1 mM, 1.5 mM, and 2 mM arsenic-treated flies. The error bar represents the means ± S.D. The significant mean difference (p < 0.05) between RI-I mean of the different samples of flies were analyzed by a one-way analysis of variance (ANOVA) (*** p < 0.0001; R-squared = 0.771).
Figure 7
Figure 7
(A) The bar graph represents the response index-II of the control flies, sucrose untreated flies, treated flies at 1 mM As, 1.5 mM As, and 2 mM As. The error bar represents the means ± S.D. The significant mean difference (p < 0.05) between RI-II mean of the different sample of flies was analyzed by a one-way analysis of variance (ANOVA) (*** p < 0.0001; R squared = 0.974. (B) The bar graph represents the learning index of arsenic-treated flies and control flies relative to untreated sucrose flies, with the error bar representing the means ± S.D. The significant mean difference (p < 0.05) between the flies’ sample mean was analyzed by a one-way analysis of variance (ANOVA) (*** p < 0.0001).
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