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. 2023 Mar 31;24(7):6575.
doi: 10.3390/ijms24076575.

Cadmium Exposure Is Associated with Behavioral Deficits and Neuroimmune Dysfunction in BTBR T+ Itpr3tf/J Mice

Mohammed M Alanazi  1 , Mushtaq A Ansari  1 , Ahmed Nadeem  1 , Sabry M Attia  1 , Saleh A Bakheet  1 , Haneen A Al-Mazroua  1 , Abdullah A Aldossari  1 , Mohammed M Almutairi  1 , Thamer H Albekairi  1 , Marwa H Hussein  1 , Mohammed A Al-Hamamah  1 , Sheikh F Ahmad  1
Affiliations

Cadmium Exposure Is Associated with Behavioral Deficits and Neuroimmune Dysfunction in BTBR T+ Itpr3tf/J Mice

Mohammed M Alanazi et al. Int J Mol Sci. .

Abstract

Autism spectrum disorders (ASD) are neurobehavioral disabilities characterized by impaired social interactions, poor communication skills, and restrictive/repetitive behaviors. Cadmium is a common heavy metal implicated in ASD. In this study, we investigated the effects of Cd exposure on BTBR T+ Itpr3tf/J (BTBR) mice, an ASD model. We looked for changes in repetitive behaviors and sociability through experiments. We also explored the molecular mechanisms underlying the effects of Cd exposure, focusing on proinflammatory cytokines and pathways. Flow cytometry measured IL-17A-, IL-17F-, IL-21-, TNF-α-, STAT3-, and RORγt-expressing CD4+ T cells from the spleens of experimental mice. We then used RT-PCR to analyze IL-17A, IL-17F, IL-21, TNF-α, STAT3, and RORγ mRNA expression in the brain. The results of behavioral experiments showed that Cd exposure significantly increased self-grooming and marble-burying in BTBR mice while decreasing social interactions. Cd exposure also significantly increased the number of CD4+IL-17A+, CD4+IL-17F+, CD4+IL-21+, CD4+TNF-α+, CD4+STAT3+, and CD4+RORγt+ cells, while upregulating the mRNA expression of the six molecules in the brain. Overall, our results suggest that oral exposure to Cd aggravates behavioral and immune abnormalities in an ASD animal model. These findings have important implications for ASD etiology and provide further evidence of heavy metals contributing to neurodevelopmental disorders through proinflammatory effects.

Keywords: BTBR mice; Th17 cells; autism spectrum disorder; behavioral studies; cadmium.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effects of oral Cd exposure on social behavior in BTBR and C57 mice. Self-grooming (A). Marble burying (B). Social proximity index (C). Social domain (D). BTBR and C57 mice were exposed to Cd (5 mg/kg/day) daily for six weeks. Control C57 and BTBR mice received saline only. All data are shown as mean ± SD (n = 10/group). Different letter indicate significant difference. * p < 0.05 vs. saline-treated C57 mice, ap < 0.05 vs. saline-treated BTBR mice; two-way ANOVA with Tukey’s test.
Figure 2
Figure 2
Effects of oral Cd exposure on IL-17A and IL-17F in BTBR and C57 mice. Spleen CD4+ T cells expressing (A) IL-17A and (B) IL-17F. RT-PCR analysis of IL-17A (C) and IL-17F (D) mRNA expression in brain tissue. (E) Representative FSC-SSC dot plots of CD4+IL-17A+ cells were obtained from each mouse spleen. BTBR and C57 mice were administered Cd (5 mg/kg/day) or saline only (control) daily for six weeks. All data are shown as mean ± SD (n = 6/group). Different letter indicate significant difference. * p < 0.05 vs. saline-treated C57 mice, ap < 0.05 vs. saline-treated BTBR mice; two-way ANOVA with Tukey’s test.
Figure 3
Figure 3
Effects of oral Cd exposure on IL-21. (A) IL-21-expressing CD4+ T cells from the spleen. (B) IL-21 mRNA expression in brain tissue. (C) Representative FSC-SSC dot plots of CD4+IL-21+ cells taken from each mouse spleen. BTBR and C57 mice were exposed to Cd (5 mg/kg/day) or saline only (control) daily for six weeks. All data are shown as mean ± SD (n = 6/group). Different letter indicate significant difference. * p < 0.05 vs. saline-treated C57 mice, ap < 0.05 vs. saline-treated BTBR mice; two-way ANOVA with Tukey’s test.
Figure 4
Figure 4
Effects of oral Cd exposure on TNF-α. (A) TNF-α-expressing CD4+ T cells from the spleen. (B) TNF-α mRNA expression in brain tissue. (C) Representative FSC-SSC dot plots of CD4+TNF-α+ cells taken from each mouse spleen. BTBR and C57 mice were orally administered Cd (5 mg/kg/day) or saline (control) daily for six weeks. All data are shown as mean ± SD (n = 6/group). Different letter indicate significant difference. * p < 0.05 vs. saline-treated C57 mice, ap < 0.05 vs. saline-treated BTBR mice; two-way ANOVA with Tukey’s test.
Figure 5
Figure 5
Effects of oral Cd exposure on STAT3. (A) STAT3-expressing CD4+ T cells from the spleen. (B) TNF-α mRNA expression in brain tissue. (C) Representative FSC-SSC dot plots of CD4+STAT3+ cells taken from each mouse spleen. BTBR and C57 mice were orally administered Cd (5 mg/kg/day) or saline (control) daily for six weeks. All data are shown as mean ± SD (n = 6/group). Different letter indicate significant difference. * p < 0.05 vs. saline-treated C57 mice, ap < 0.05 vs. saline-treated BTBR mice; two-way ANOVA with Tukey’s test.
Figure 6
Figure 6
Effects of oral Cd exposure on RORγt. (A) RORγt-expressing CD4+ T cells from the spleen. (B) RORγ mRNA expression in brain tissue. (C) Representative FSC-SSC dot plots of CD4+RORγt+ cells taken from each mouse spleen. BTBR and C57 mice were orally administered Cd (5 mg/kg/day) or saline only (control) daily for six weeks. All data are shown as mean ± SD (n = 6/group). Different letter indicate significant difference. * p < 0.05 vs. saline-treated C57 mice, ap < 0.05 vs. saline-treated BTBR mice; two-way ANOVA with Tukey’s test.
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References

    1. American Psychiatric Association . Diagnostic and Statistical Manual of Mental Disorders. 5th ed. American Psychiatric Publishing; Washington, DC, USA: 2015. Autism Spectrum Disorder.
    1. Sharma S.R., Gonda X., Tarazi F.I. Autism Spectrum Disorder: Classification, diagnosis and therapy. Pharmacol. Ther. 2018;190:91–104. doi: 10.1016/j.pharmthera.2018.05.007. - DOI - PubMed
    1. Freeman K. Nutrient protection against arsenic toxicity: Folate, cysteine support methylation in children. Environ. Health Perspect. 2009;117:A211. doi: 10.1289/ehp.117-a211b. - DOI - PMC - PubMed
    1. Chen H., Ahn R., Van den Bossche J., Thompson D.H., O’Halloran T.V. Folate-mediated intracellular drug delivery increases the anticancer efficacy of nanoparticulate formulation of arsenic trioxide. Mol. Cancer Ther. 2009;8:195563. doi: 10.1158/1535-7163.MCT-09-0045. - DOI - PMC - PubMed
    1. Hall M.N., Liu X., Slavkovich V., Ilievski V., Pilsner J.R., Alam S., Factor-Litvak P., Graziano J.H., Gamble M.V. Folate, Cobalamin, Cysteine, Homocysteine, and Arsenic Metabolism among Children in Bangladesh. Environ. Health Perspect. 2009;117:825–831. doi: 10.1289/ehp.0800164. - DOI - PMC - PubMed

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Grants and funding

This research was funded by Deputyship for Research & Innovation, Ministry of Education in Saudi Arabia (IFKSURG-2-210).
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