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...

A number of mechanisms have been proposed to contribute the selective neuronal cell loss observed during Alzheimer disease (AD). These include formation and accumulation amyloid-β (Aβ)-containing plaques, neurofibrillary tangles (NFTs), inflammatory processes mediated by astrocytes microglia. Neuronal responses such insults in AD brain increased protein levels immunoreactivity for kinases known regulate cycle progression. One downstream target these regulatory proteins, Retinoblastoma...

Mitochondria have been identified as targets of the neurotoxic actions zinc, possibly through decreased mitochondrial energy production and increased reactive oxygen species accumulation. It has hypothesized that impairment trafficking may be a mechanism neuronal injury. Here, we report elevated intraneuronal zinc impairs trafficking. At concentrations just sufficient to cause injury, rapidly inhibited movement without altering morphology. Zinc chelation initially restored movement, but...

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...

Abstract Estrogens influence differentiation, growth and function of neurons, but less is known their effects on glia. In our experiments reported here, the ovarian steroid, 17 β ‐estradiol, “designer”, non‐steroidal estrogen, tamoxifen, effectively protected C‐6 glioma 2B clone cells from cytotoxicity excitatory neurotransmitter, glutamate. Exposure these to 10–20 mM glutamate induced 61–78% cell death. Pre‐treatment with 0.01 estradiol or 2 μM tamoxifen significantly reduced...

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...

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...

Polybrominated diphenyl ethers (PBDE) are a class of flame retardants with wide industrial and domestic use that known to disrupt endocrine function. Because their long half‐life in the environment ability bioaccumulate fatty tissues, there is significant interest understanding long‐term effects on human health. Our research investigates particular PBDE, BDE‐47, steroid‐producing adrenocortical carcinoma cell line (HAC15). We found BDE‐47 exposure HAC15 cells significantly increases...

Elevated circulating aldosterone levels are associated with hypertension, thrombosis formation, cardiac hypertrophy, and congestive heart failure. Aldosterone secretion is regulated by angiotensin II (Ang II), potassium, adrenocorticotropic hormone (ACTH). Calcium required for the stimulation of these steroidogenic stimuli. Despite importance calcium in steroidogenesis, relationship between intracellular Ca 2+ homeostasis steroid production adrenal cortex remains incompletely understood. We...

A previous body of work in bovine and rodent models shows that cholinergic agonists modulate the secretion steroid hormones from adrenal cortex. In this study we used live-cell Ca2+ imaging to investigate activity HAC15 human adrenocortical carcinoma cell line. The carbachol acetylcholine triggered heterogeneous oscillations were strongly inhibited by antagonists with high affinity for M3 muscarinic receptor subtype, while preferential block M1 or M2 receptors was less effective. Acute...

Intracellular calcium fluctuations may play a role in the control of corticosteroid synthesis and secretion by adrenal cortex. Using live-cell imaging with calcium-sensitive fluorescent probe Fluo-4 human adrenocortical carcinoma cell line (HAC15), we previously demonstrated that activation M3 muscarinic receptors stimulates intracellular calcium oscillations, enhances aldosterone production, increases expression of CYP11B2 (1).  To further understand autonomic...

<sec> <title>BACKGROUND</title> 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. </sec> <title>OBJECTIVE</title> We evaluated hypnotic medications, commonly used their therapeutic efficacy...

Abstract The role of autonomic receptors in the regulation adrenal cortex is poorly understood. We recently showed that activation M3 muscarinic stimulates intracellular calcium oscillations, aldosterone production, and expression CYP11B2 (1). present study explores relationship between adrenergic corticosteroid production. Using live-cell fluorescence imaging HAC15 adrenocortical cells with calcium-sensitive probe Fluo-4, we have shown stimulation endogenous, non-selective agonist...

Mitochondria may be both the source and target of oxidative stress in neurodegenerative disease. In models excitotoxicity, neuronal injury is triggered by influx calcium into neurons then mitochondria. Our studies suggest that an important consequence this movement generation ROS Studies with isolated mitochondria enhance mitochondria, especially when complex I impaired. However, these are complicated a lack specificity detection methods like Amplex Red. One feature their within neurons. We...