Edited by Louis J. Ignarro, University of California, Los Angeles School Medicine, Angeles, CA, and approved August 27, 2001 (received for review May 16, 2001)

Abstract: The membrane‐permeant oxidizing agent 2,2′‐dithiodipyridine (DTDP) can induce Zn 2+ release from metalloproteins in cell‐free systems. Here, we report that brief exposure to DTDP triggers apoptotic cell death cultured neurons, detected by the presence of both DNA laddering and asymmetric chromatin formation. Neuronal was blocked increased extracellular potassium levels, tetraethylammonium, broad‐spectrum cysteine protease inhibitor butoxy‐carbonyl‐aspartate‐fluoromethylketone....

We hypothesized that metallothionein (MT), a cysteine-rich protein with strong affinity for Zn 2+ , plays role in nitric oxide (NO) signaling events via sequestration or release of by the unique thiolate clusters protein. Exposing mouse lung fibroblasts (MLF) to NO donor S-nitrosocysteine resulted 20–30% increases fluorescence -specific fluorophore Zinquin were rapidly reversed chelator N,N,N′, N′-tetrakis-(2-pyridylmethyl)ethylenediamine. The absence NO-mediated increase labile MLF from MT...

Iron is an essential element for cells but may also be important cytotoxin. However, very little known about iron transport, redox status, or toxicity specifically inside cells. In this study, we exploited the sensitivity of fura-2 to quenching by ferrous (Fe 2+ ) detect intracellular free ([Fe ] i in neurons, astrocytes, and oligodendrocytes primary culture. All cell types exposed Fe presence ionophore pyrithione rapidly accumulated a similar extent. The heavy-metal chelators bipyridyl...

The emergence of zinc as a potent neurotoxin has prompted the development techniques suitable for measurement intracellular free ([Zn²⁺]_i) in cultured cells. Accordingly, new family Zn²⁺-sensitive fluorophores become available. Using ionophore-induced elevations [Zn²⁺]_i neurons, we measured [Zn²⁺]_i-induced changes novel dyes FuraZin-1 and FluoZin-2 compared them with established...

Abstract Current evidence suggests that zinc kills neurons by disrupting energy production, specifically inhibiting mitochondrial function. However it is unclear if the inhibitory effect requires accumulation, and so, precisely how enters mitochondria. Here, using fluorescence microscopy to visualize individual rat brain mitochondria, we detected matrix uptake fluorophore FluoZin‐3. Fluorescence increased rapidly in mitochondria treated with micromolar free zinc, was quickly returned...

Liberation of zinc from intracellular stores contributes to oxidant-induced neuronal injury. However, little is known regarding how endogenous oxidant systems regulate free ([Zn(2+)](i)). Here we simultaneously imaged [Ca(2+)](i) and [Zn(2+)](i) study acute changes in cultured rat forebrain neurons after glutamate receptor activation. Neurons were loaded with fura-2FF FluoZin-3 follow [Zn(2+)](i), respectively. treated (100 microM) for 10 min gave large Ca(2+) responses that did not recover...

Abstract Excessive accumulation of the heavy metal zinc is cytotoxic. As a consequence, cellular vulnerability to zinc‐induced injury may be regulated by abundance proteins that maintain intracellular free concentrations ([Zn 2+ ] i ). In this study, we overexpressed zinc‐binding protein metallothionein‐II (MT) in astrocytes assess its impact as (1) an acute buffering mechanism, and (2) oxidant‐releasable pool. Overexpression MT primary astrocyte cultures was accomplished using adenoviral...

Patient online drug reviews are a resource for other patients seeking information about the practical benefits and drawbacks of therapies. may also serve as source postmarketing safety data that more user-friendly than regulatory databases. However, reliability has been questioned, because they do not undergo professional review lack means verification.We evaluated hypnotic medications, commonly used their therapeutic efficacy is particularly amenable to patient self-evaluation. Our primary...

Excessive accumulation of intracellular free zinc is lethal to neurons. The precise mechanism by which kills neurons remains unclear, however, a number reports suggest that may impair neuronal metabolism. present study examined how affects the activity isolated rat brain mitochondria. Using spectrofluorometry and polarography, we determined effects on mitochondrial membrane potential (Δψ), reactive oxygen species (ROS) generation, respiration. In mitochondria respiring either glutamate +...

Abstract The understanding of cellular Cd 2+ accumulation and toxicity is hampered by a lack fluorescent indicators selective for intracellular free ([Cd ] i ). In this study, we used depolarized MIN6 mouse pancreatic beta cells as model evaluating [Cd detection with commercially available probes, most which have been traditionally to visualize [Ca [Zn . We trialed panel 12 probes including fura-2, FluoZin-3, Leadmium Green, Rhod-5N, indo-1, Fluo-5N, others. found that the probe FluoZin-3...

Fluorescent probes such as fura-2 and its relatives mag-fura-2 fura-2FF are used to detect changes in intracellular ions Ca2 Mg2 . However, they sufficiently non-selective record other species Zn2 , Cd2 Fe2 transition metals. In the present study we have characterized responses of these three dyes a buffered salt solution using spectrofluorophotometry. Fura-2, generally reacted increasing concentrations or with shift excitation spectra. We found that for given probe spectra were broadly...

Cadmium is a common industrial pollutant, but the study of cadmium cytotoxicity difficult in part due to absence probe that can detect living cells without confounds contaminating metals. Leadmium Green (LG) commercially available, small molecule, fluorescent advertised as detector intracellular and lead. LG has been used monitor sequestration plant cells, heavy metal export protozoan, lead absorption by vascular endothelial cells. However very little information available regarding its...