Chronic high dose intraperitoneal bisphenol A (BPA) induces substantial histological and gene expression alterations in rat penile tissue without impairing erectile function.

INTRODUCTION: Bisphenol A (BPA), released from plastics and dental sealants, is a suspected endocrine disruptor and reproductive toxicant. In occupationally exposed workers, BPA has been associated with erectile dysfunction (ED). AIMS: To determine whether long-term exposure to high doses of BPA in the rat affects serum levels of testosterone (T) and estradiol (E2), and induces corporal histopathology and resultant ED. METHODS: Young rats were injected intraperitoneal (IP) injection daily with BPA at 25 mg/kg/day or vehicle (n = 8/group). Erectile function was measured at 3 months by cavernosometry and electrical field stimulation (EFS). BPA was assayed in serum, urine, and penile tissue, and serum T and E2 were determined. Quantitative Masson trichrome, terminal deoxynucleotidyl transferase dUTP nick end labeling, Oil Red O, immunohistochemistry for calponin, α-smooth muscle actin, and Oct 4 were applied to penile tissue sections. Protein markers were assessed by Western blots and 2-D minigels, and RNA by DNA microarrays. MAIN OUTCOME MEASURES: Erectile function, histological, and biochemical markers in corporal tissue. RESULTS: In the BPA-treated rats, total and free BPA levels were increased in the serum, urine, and penile tissue while serum T and E2 levels were reduced. In addition, the corpora cavernosa demonstrated a reduction in smooth muscle (SM) content, SM/collagen ratio, together with an increase in myofibroblasts, fat deposits, and apoptosis, but no significant change in collagen content or stem cells (nuclear/perinuclear Oct 4). In the penile shaft, BPA induced a downregulation of Nanog (stem cells), neuronal nitric oxide synthase (nitrergic terminals), and vascular endothelial growth factor (angiogenesis), with genes related to SM tone and cytoskeleton upregulated 5- to 50-fold, accompanied by changes in the multiple protein profile. However, both cavernosometry and EFS were unaltered by BPA. CONCLUSIONS: While rats treated chronically with a high IP dose of BPA developed hypogonadism and a corporal histo- and molecular-pathology usually associated with ED, no changes were detected in erectile function as measured by EFS and cavernosometry. Further studies using alternate routes of BPA administration with various doses and length of exposure are needed to expand these findings.

Excretion of PFOA and PFOS in male rats during a subchronic exposure.

Perfluorinated compounds (PFCs), a class of synthetic surfactants that are widely used, have become global environmental contaminants because of their high persistence and bioaccumulation. An increasing number of studies have described the pharmacokinetics of PFCs following in vivo exposure, however, few papers have focused on the excretion of these compounds during a period of consecutive exposure. In this study, the excretions of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) in male Sprague-Dawley rats gavaged consecutively for 28 days were investigated and compared. The faster elimination rate in urine compared to feces indicated that urinary excretion is the primary clearance route in rats for either PFOA or PFOS. During the first 24 h after administration of PFOA (5 and 20 mg/kg body weight/day), about 24.7-29.6% of the oral dose was excreted through urine and feces, while for PFOS, the excretion amounts were only 2.6-2.8% of the total gavaged doses (5 and 20 mg/kg body weight/day). The excretion rates of both PFCs increased with increasing exposure doses. The higher elimination rate of PFOA through excretion indicated its lower accumulation in rats, thus inducing possible lower toxicities compared to PFOS.

Studies on the toxicological effects of PFOA and PFOS on rats using histological observation and chemical analysis.

As an emerging class of environmentally persistent and bioaccumulative contaminants, perfluorinated compounds (PFCs), especially perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS), have been ubiquitously found in the environment. Increasing evidence shows that the accumulated levels of PFCs in animals and the human body might cause potential impairment to their health. In the present study, toxicological effects of PFOA and PFOS on male Sprague-Dawley rats were examined after 28 days of subchronic exposure. Abnormal behavior and sharp weight loss were observed in the high-dose PFOS group. Marked hepatomegaly, renal hypertrophy, and orchioncus in treated groups were in accordance with the viscera-somatic indexes of the liver, kidney, and gonad. Histopathological observation showed that relatively serious damage occurred in the liver and lung, mainly including hepatocytic hypertrophy and cytoplasmic vacuolation in the livers and congestion and thickened epithelial walls in the lungs. PFOA concentrations in main target organs were in the order of kidney > liver > lung > (heart, whole blood) > testicle > (spleen, brain), whereas the bioaccumulation order for PFOS was liver > heart > kidney > (whole blood) > lung > (testicle, spleen, brain). The highest concentration of PFOA detected in the kidney exposed to 5 mg/kg/day was 228+/-37 microg/g and PFOS in the liver exposed to 20 mg/kg/day reached the highest level of 648+/-17 microg/g, indicating that the liver, lung, and kidney might serve as the main target organs for PFCs. Furthermore, a dose-dependent accumulation of PFOS in various tissues was found. The accumulation levels of PFOS were universally higher than PFOA, which might explain the relative high toxicity of PFOS. The definite toxicity and high accumulation of the tested PFCs might pose a great threat to biota and human beings due to their widespread application in various fields.

Effects of perfluorooctane sulfonate on ion channels and glutamate-activated current in cultured rat hippocampal neurons.

Concern on an emerging persistent contaminant, perfluorooctane sulfonate (PFOS), is increasingly growing. Although the fate, transport, distribution and bioaccumulation of PFOS have been documented, its toxicological effects especially neurotoxicity remain largely unknown. In this study, the effects of PFOS on ion channels including potassium and sodium channels and exogenous glutamate-activated current in cultured rat hippocampal neurons were examined, based on whole-cell patch-clamp recording. PFOS markedly increased two subtypes of potassium currents, including transient outward current and delayed rectifier current, at doses over 10µM. PFOS did not affect the amplitude of sodium current at all administrated doses (1, 10 or 100µM) but clearly shifted the activation current-voltage curve toward negatively potential. Further, PFOS significantly altered the glutamate-activated current at all doses. Taken together these findings indicated that PFOS disturbs the neuronal physiological processes, which revealed the damage of this pollutant to nerve system and will be helpful for further exploration to its underlying mechanism.

Acute enhancement of synaptic transmission and chronic inhibition of synaptogenesis induced by perfluorooctane sulfonate through mediation of voltage-dependent calcium channel.

Perfluorooctane sulfonate (PFOS) is a persistent and bioaccumulative pollutant ubiquitous in wildlife and humans. Although the distribution and fate of PFOS have been widely studied, its potential neurotoxicity remains largely unknown. In the present study, the acute and chronic effects of PFOS on the development and synaptic transmission of hippocampal neurons was examined. Perfusion with PFOS markedly increased the frequency of miniature postsynaptic currents (mPSCs) and slightly elevated the amplitude of mPSCs in cultured hippocampal neurons. Perfusion with PFOS also increased the amplitude of field excitatory postsynaptic potentials (fEPSPs) recorded in the CA1 region of hippocampal slices. Both of these effects were largely blocked by the L-type Ca2+ channel antagonist nifedipine. Further studies showed that PFOS enhanced inward Ca2+ currents and increased intracellular Ca2+ in cultured neurons; these effects were also substantially inhibited by nifedipine. Moreover, prolonged treatment with PFOS moderately inhibited neurite growth and dramatically suppressed synaptogenesis in cultured neurons in a nifedipine-sensitive manner. Thus, through enhancement of Ca2+ channels, PFOS may exhibit both acute excitotoxic effects on synaptic function and chronically inhibit synaptogenesis in the brain.

Interaction of methylmercury and selenium on the bioaccumulation and histopathology in medaka (Oryzias latipes).

Interaction of methylmercury and selenium in medaka (Oryzias latipes) on bioaccumulation of pollutants and histopathological changes in liver and gill were studied. Juvenile medaka fish were submitted to a series of waterborne methylmercury chloride (MMC), sodium selenite (Na(2)SeO(3)) and their mixture for 210 days, respectively. The methylmercury (MeHg) and selenium contents in the whole body of medaka were determined. The dose- and time-dependent increase of MeHg and selenium contents in medaka were observed. Histopathological changes, such as edema, vacuoles, pyknotic nucleus, and telangiectasis, could clearly be observed in the slices from the exposed medaka's liver and gill. Concurrent exposure to MMC and Na(2)SeO(3) showed the increased selenium accumulation. When the exposure molar ratio of MeHg:Se was about 1, the interaction between MeHg and selenium offered a limited protection against the serious intoxication of both MMC and Na(2)SeO(3) to medaka.

[Determination of methylmercury in biological and sediment samples by capillary gas chromatography on-line coupled with atomic fluorescence spectrometry].

A comprehensive method for the determination of methylmercury by capillary gas chromatography (GC) on-line coupled with atomic fluorescence spectrometry (AFS) was developed. The hyphenation system was carefully designed and the operation conditions such as injector temperature, carrier gas flow rate, make-up gas flow rate and argon flow rate were optimized in detail. The absolute detection limits (3sigma) were 0.005 ng of Hg for both methylmercury (MMC) and ethylmercury (EMC). The relative standard deviations (RSD, n = 5) for MMC and EMC at 10 ng x mL(-1) of Hg levels were 2.5% and 1.3%, respectively. The method was evaluated by analyzing a certified reference material (DORM-2), and the MMC concentration determined was in good agreement with the certified value. In addition, the recoveries of MMC in two sediment samples were 70% and 77%, respectively. The proposed method was accurate, sensitive and suitable for the determination of MMC in biological and sediment samples.

Methylmercury accumulation, histopathology effects, and cholinesterase activity alterations in medaka (Oryzias latipes) following sublethal exposure to methylmercury chloride.

A series of specific toxicological effects including bioaccumulation of the pollutant, histological changes and influences on cholinesterase (ChE) activities were examined in the adult Japanese medaka after the exposure to graded sublethal concentrations (40, 20, 10, 5, 2.5ngHg/mL) of methylmercury chloride (MMC). Methylmercury (MeHg) contents in the exposed medaka tissues ranged from 0.03 to 64.4µgHg/g (wet weight, w.w.). High concentrations of MeHg were accumulated in the liver and brain, while the concentrations in muscle and fat were relatively low. A dose-dependent and exposure time-dependent increase of MeHg contents in tissues was observed. Histopathological changes, such as oedema, vacuolization, pyknotic nucleus, telangiectasis, and degenerative sperm, can clearly be observed in the slices from the liver, gill, and male gonad of the exposed medaka. Inhibition of ChE activity was common in the exposed fish's brain, liver, gill, and muscle. The serious intoxication of MMC to medaka was definitely demonstrated herein.