Effects of copper nanoparticles on oxidative stress genes and their enzyme activities in common carp (Cyprinus carpio)

Figures & data

Table I. List of gene primers used in this study (Scown et al. 2010; Zhang et al. 2016; Fazelan et al. 2020).

Primer	Primer sequence (5´-3´)	Annealing Temperature	Amplicon Length (bp)
cyp1a2F	CTTCCGCCATATTGTCGTATC	52.4°C	135
cyp1a2R	CCACCACCTGCCCAAAC	51.9°C
hsp70F	CTGCTGCTGCTGGATGTG	52.6°C	25
hsp70R	GCTGGTTGTCGGAGTAAGTG	53.8°C
sodF	TGAGCTGTCGGAAGCCATCAAG	56.7°C	150
sodR	TTGGTTCCCACATGCAGCAATCC	57.1°C
catF	AGACGACACCCATCGCTGTTCG	58.6°C	120
catR	AAGGTCCCAGTTGCCCTCATCG	58.6°C
gpxF	GCTCCATTCGCAGTATTC	48.0°C	50
gpxR	TCCTTCCCATTCACATCC	48.0°C
ef1aF	CTTCGTCCCAATTTCTG	44.6°C	100
ef1aR	ACCGTTAGCATTACCCT	44.6°C
bactinF	CCTGTATGCCAACACCGTGCTG	58.6°C	100
bactinR	CTTCATGGTGGAGGGAGCAAGG	58.6°C

Figure 1. Effects of Cu, Cu-NPs or CuO (concentration 1 µg L⁻¹) on enzymatic activity and transcribed level of cyp1a (a, b), hsp70 (c, d), sod (e, f), cat (g, h) and gpx (i, j). Relative expression was normalized by bactin and ef1a as reference transcripts. Vertical bars indicate ± SD. Duncan’s multiple range test was used to test for significance of differences (p ≤ 0.05) in enzymatic activity. Statistical analyses of gene expression were performed using a post hoc Tukey’s (HSD) test with a confidence interval of 0.05 and a confidence level of 95%. Differences between the mean values were considered to be significant at p < 0.01 or p < 0.05. Mean values with different letters (a–d) are significantly different (p < 0.05).

Figure 2. Effects of Cu, Cu-NPs or CuO (concentration 1 µg L⁻¹) on gene expression reveiled as protein level of CYP1A (a) and HSP70 (b). Vertical bars indicate ± SD. Duncan’s multiple range test was used to test for significance of differences (p ≤ 0.05). Mean values with different letters (a–b) are significantly different (p < 0.05).

Figure 3. Effect of Cu, Cu-NPs or CuO-NPs (concentration 1 µg L⁻¹) on Cu/Zn-SOD (a) and CAT (b) isoenzyme activity. Vertical bars indicate ± SD. Duncan’s multiple range test was used to test for significance of differences (p ≤ 0.05). Mean values with different letters (a–d) are significantly different (p < 0.05).

Figure 4. Effect of Cu, Cu-NPs or CuO-NPs (concentration 1 µg L⁻¹) on superoxide anion (O₂^•−) (a) and hydrogen peroxide (H₂O₂) (b) levels. Vertical bars indicate ± SD. Duncan’s multiple range test was used to test for significance of differences (p ≤ 0.05). Mean values with different letters (a-d) are significantly different (p < 0.05).

Scown TM, Santos EM, Johnston BD, Gaiser B, Baalousha M, Mitov S, Lead JR, Stone V, Fernandes TF, Jepson M, van Aerle R, Tyler CR. 2010. Effects of aqueous exposure to silver nanoparticles of different sizes in rainbow trout. Toxicological Sciences 115(2):521–534. DOI: 10.1093/toxsci/kfq076.

 PubMed Web of Science ®Google Scholar

Zhang W, Jia Y, Ji X, Zhang R, Liang T, Du Q, Chang Z. 2016. Optimal reference genes in different tissues, gender, and gonad of Yellow River Carp (Cyprinus carpio var) at various developmental periods. Pakistan Journal of Zoology 48(6).

 Web of Science ®Google Scholar

Fazelan Z, Hoseini SM, Yousefi M, Khalili M, Hoseinifar SH, Van Doan H. 2020. Effects of dietary eucalyptol administration on antioxidant and inflammatory genes in common carp (Cyprinus carpio) exposed to ambient copper. Aquaculture 520:734988. DOI: 10.1016/j.aquaculture.2020.734988.

 Web of Science ®Google Scholar

Data availability statement

The datasets generated and analyzed during the current study are available from the corresponding author on reasonable request.