Mercury Impairment of Mouse Thymocyte Survival in Vitro: Involvement of Cellular Thiols

Abstract

Heavy metals are well known to be able to induce immunotoxicity, but comparative metal studies related to apoptosis have not been conducted. In the present study, the effects of arsenic, cadmium, gold, lead, manganese, and mercury on thymocytes from BALB/c mice were analyzed. Thymic cells were cultured for 3-24 h in vitro in the absence or presence of metal, and markers of apoptosis or cell death, including annexin V binding, DNA loss/oligonucleosomal fragmentation, 7-amino-actinomycin D uptake (loss of impermeance), changes of the mitochondrial membrane potential (JC-1 fluorescence), and Western analysis of cellular thiols, were assayed. Mercury (Hg) was the only metal shown to be consistently toxic with the dose and times utilized. Cadmium (Cd) was the only other metal tested that also produced some significant level of DNA loss; however, the induction of apoptosis by Cd was not as consistent as that observed with Hg. When Hg was added with 2-mercaptoethanol (2-ME), Hg produced greater toxicity. Endogenous DNA synthesis by thymocytes was immediately inhibited by Hg and Hg + 2-ME. The Hg + 2-ME-induced apoptosis appeared to be associated with altered levels of cellular thiols, in that glutathione (GSH) depletion was significant in comparison to the non-metal control and Hg alone. The increased Hg-induced toxicity in the presence of 2-ME likely was due to the ability of 2-ME to enhance (10- to 20-fold) the cellular uptake of Hg. Western analysis with biotin maleimide demonstrated that Hg + 2-ME and to a lesser extent the positive control dexamethasone eliminated many reactive thiols; the major thiol-reactive protein still reactive with the maleimide probe had an approximate Molecular Mass of 45 kD. Surprisingly, Hg alone enhanced the expression of this thiol-expressing protein, which by Mass Spectrometry (MS)/MS analysis was shown to be beta-actin. Hg also produced the appearance of yet to be identified new proteins. Based on the results with Hg + 2-ME, it is suggested that numerous protein thiols participate in maintenance of cell survival and their loss is associated with apoptosis. The increased expression of new thiol-reactive proteins or thiol-reactive proteins with altered electrophoretic profiles needs to be further investigated. However, the enhanced toxicity attributed to Hg + 2-ME suggests that increased intracellular oxidative stress, observed as increased depletion of GSH, is responsible for the accelerated cell death.

Inhibition of cadmium- induced genotoxicity and histopathological changes in Nile tilapia fish by Egyptian and Tunisian montmorillonite clay

“…Cadmium was known by their bioaccumulation and caused harmful effects in different levels of the trophic chain (Stoeppler, 1991). It is well known as carcinogen (Banerjee and Flores-Rozas, 2005), teratogenic (Hovland et al, 2000), caused sterility (Bench et al, 1999), nephrotoxic (Ahmed and Abdel-Wahhab, 2000;Cai et al, 2001), hepatotoxic (Horiguchi et al, 2000;Ahmed and Abdel-Wahhab, 2000), genotoxic and apoptotic (Kim et al, 2005;Mondal et al, 2005) and also inducing pancreatic activity changes (Shimada et al, 2000). For this fact, excessive exposure to Cd results in diseases and occasionally death (Othumpangat et al, 2005).…”

Mouse Monocytes (RAW CELLS) and the Handling of Cysteine and Homocysteine S-Conjugates of Inorganic Mercury and Methylmercury

“…Recent data from mouse thymocytes exposed to heavy metals such as mercury, cadmium, arsenic, gold, lead, or manganese show that mercury was the only metal that consistently induced toxic effects in these cells (Mondal et al, 2005). Moreover, cellular intoxication by mercury was increased in the presence of 2-metacaptoethanol (2-ME), which apparently enhanced the cellular uptake of inorganic mercury (Hg 2+ ) by 10-to 20-fold.…”

Correlations Between Gene Expression and Mercury Levels in Blood of Boys With and Without Autism

“…It is interesting that most of these correlations are negative. Chronic exposure to low concentrations of heavy metals, including mercury, causes immune dysfunction related in large part to apoptosis of leukocytes (Pollard and Hultman 1997; Zelikoff and Gardner 1996; Laiosa et al 2007; Madureira et al 2007; Ziemba et al 2005; Mondal et al 2005; McCabe et al 2003, 2005; Cunha et al 2004; Sarmento et al 2004; Kuo and Lin-Shiau 2004; Colombo et al 2004; Field et al 2003). Both inorganic and organic mercurials cause human T-cell apoptosis with mitochondria being the target organelle for the induction of cell death (Shenker et al 1999).…”