Ascorbic Acid and Graphene Oxide Exposure in the Model Organism Acheta domesticus Can Change the Reproduction Potential

Abstract

The use of nanoparticles in the industry carries the risk of their release into the environment. Based on the presumption that the primary graphene oxide (GO) toxicity mechanism is reactive oxygen species production in the cell, the question arises as to whether well-known antioxidants can protect the cell or significantly reduce the effects of GO. This study focused on the possible remedial effect of vitamin C in Acheta domesticus intoxicated with GO for whole lives. The reproduction potential was measured at the level of Vitellogenin (Vg) gene expression, Vg protein expression, hatching success, and share of nutrition in the developing egg. There was no simple relationship between the Vg gene's expression and the Vg protein content. Despite fewer eggs laid in the vitamin C groups, hatching success was high, and egg composition did not differ significantly. The exceptions were GO20 and GO20 + Vit. C groups, with a shift in the lipid content in the egg. Most likely, ascorbic acid impacts the level of Vg gene expression but does not affect the production of Vg protein or the quality of eggs laid. Low GO concentration in food did not cause adverse effects, but the relationship between GO toxicity and its concentration should be investigated more thoroughly.

Keywords: Acheta domesticus; ascorbic acid; graphene oxide; vitellogenin.

Figure 1

Vg gene expression levels in A. domesticus fat body on the 5th and 15th days of adult life. Data were shown as a relative expression compared to β-actin and expressed as means ± SE. Abbreviations: Control—animals fed uncontaminated food; Vit. C.—animals fed with Vitamin C in the food; GO20 + Vit. C.—animals fed with graphene oxide and Vitamin C; GO20—animals fed with graphene oxide; significant differences were measured using ANOVA (Fisher test; p < 0.05); different letters denote differences among the experimental groups within time points.

Figure 2

Total Vg protein content in the fat body of A. domesticus on the 5th and 15th days of adult life. Abbreviations: Control—animals fed uncontaminated food; Vit. C.—animals fed with Vitamin C in the food; GO20 + Vit. C.—animals fed with graphene oxide and Vitamin C in the food; GO20—animals fed with graphene oxide in the food; significant differences were measured using ANOVA (Fisher test; p < 0.05); different letters denote differences among the experimental groups and time points.

Figure 3

Semi-quantitative analysis of Vg protein in the fat body of A. domesticus on the 5th and 15th days of adult life. The graph presents the precursor’s Vg expression (~200 kDa) (A,B) and the subunits 130 kDa (C,D), 97 kDa (E,F). Abbreviations: Control—animals fed uncontaminated food; Vit. C.—animals fed with Vitamin C in the food; GO20 + Vit. C.—animals fed with graphene oxide and Vitamin C; GO20—animals fed with graphene oxide; Expression measured as band density compared to the reference (control, day 5th). All the groups were compared to controls presented as 100% (red line).

Figure 4

Egg laying (48 h) and hatching success of A. domesticus. (A) the average number of eggs laid per female (dark grey), the average number of larvae enclosed per female (light grey), and (B) the average total hatching success measured as a percent of enclosed eggs to the total number of laid eggs in the experimental group. Abbreviations: Control—animals fed uncontaminated food; Vit. C.—animals fed with Vitamin C in the food; GO20 + Vit. C.—animals fed with graphene oxide and Vitamin C; GO20—animals fed with graphene oxide; significant differences were measured using ANOVA (Fisher test; p < 0.05); different letters denote differences among the experimental groups (lower case: eggs/female, capital letters: larvae/female).

Figure 5

Share of major energetic components (lipids, glucose, glycogen, and total protein content) in the eggs of A. domesticus females collected on the 5th and 15th days of adult life. Abbreviations: Control—animals fed uncontaminated food; Vit. C.—animals fed with Vitamin C in the food; GO20 + Vit. C.—animals fed with graphene oxide and Vitamin C; GO20—animals fed with graphene oxide.

Figure 6

Major energetic components: lipids (A), glucose (B), glycogen (C), and total protein content (D) in the eggs of A. domesticus females collected on the 5th and 15th day of adult life. Abbreviations: Control—animals fed uncontaminated food; Vit. C.—animals fed with Vitamin C in the food; GO20 + Vit. C.—animals fed with graphene oxide and Vitamin C; GO20—animals fed with graphene oxide; significant differences were measured using ANOVA (Fisher test; p < 0.05); different letters denote differences among the experimental groups (small letters for the 5th day, and capital letters for the 15th day); an asterisk (*) and hashtag (#) show differences between corresponding groups on days 5th and 15th.

Figure 7

Image of graphene oxide (A) SEM Magnification: 10,000×; scale bar 10 µm; (B) AFM.