Biomedical subjects

Frode Fonnum

Publications and source records attributed to Frode Fonnum.

29 records · Page 2Linked to original sources

PubMed2003

D-Serine alleviates retrograde amnesia of a visual discrimination task in rats with a lesion of the perirhinal cortex.

D-Serine has been suggested to be a potent endogenous glycine-site agonist on the N-methyl-D-aspartate receptor, thereby having a potential role in the process of learning and memory. In rats, perirhinal cortex (PC) constitutes a particularly important structure for mnemonic processing, and damage to this area induces both anterograde and retrograde amnesia. In the present work, we show that intraperitoneal administration of 1000 mg/kg D-serine immediately after bilateral lesion of PC produced complete restoration of retrograde memory in rats, measured by a visual brightness discrimination task, while a higher dose (3000 mg/kg) did not show any reliable effect. Uptake of the drug into the brain was confirmed using high performance liquid chromatography (HPLC).

Amnesia, Retrograde↗

PubMed2003

Involvement of the extracellular signal regulated kinase pathway in hydrocarbon-induced reactive oxygen species formation in human neutrophil granulocytes.

In the present study we have examined the effects of hydrocarbons on the formation of reactive oxygen species (ROS) in human neutrophil granulocytes in vitro. We found that hydrocarbons induce ROS formation in a concentration-dependent manner and that the ROS-inducing potency increases with increasing number of carbon atoms in the structure. In general, aromatic hydrocarbons were less potent inducers of ROS than aliphatic and cyclic hydrocarbons. The most potent compound in each group, t-butylcyclohexane, n-decane, and n-butylbenzene, were chosen for mechanistic studies. ROS formation was inhibited by the MEK1/2 inhibitor U0126, the tyrosine kinase inhibitor erbstatin-A, and the phosphatidylinositol-3 kinase inhibitor wortmannin. The involvement of the ERK1/2 pathway was confirmed by Western blot analysis of phosphorylated ERK1/2. The study revealed only small differences in the mechanisms involved for the three compounds. The responses were not affected by Pertussis toxin, indicating that Gi-protein coupled receptors are not involved in neutrophil activation after hydrocarbon exposure. Based on these findings we propose a mechanism involving tyrosine kinases, PI3 kinase, and the ERK1/2 pathway, leading to activation of the NADPH oxidase and production of ROS in neutrophils stimulated by organic solvents.

Adult↗

PubMed2003

Evaluation of the probes 2',7'-dichlorofluorescin diacetate, luminol, and lucigenin as indicators of reactive species formation.

This study attempts to provide a critical assessment of three different common approaches to identifying teactive species formed in biological systems: the 2',7'-dichlorofluorescin diacetate (DCFH-DA) assay, and the luminol- and lucigenin-amplified chemiluminescence assays. There have been several contradictory reports about the specificity of these methods. Our results show that DCFH is oxidized to the fluorescent compound 2',7'-dichlorofluorescin (DCF) in human neutrophils exposed to the following compounds: Aroclor (A)1242, hydrogen peroxide (H(2)O(2)), nitric oxide (NO), and FeSO(4). Use of a cell-free DCFH system showed increased formation of DCF by peroxynitrite (ONOO(-)), horseradish peroxidase (HRP) alone, and HRP in combination with H(2)O(2), FeSO(4) alone, and a mixture of FeSO(4) and H(2)O(2). The hydroxyl radical (z.rad;OH) scavenger formate and the iron ion chelator deferoxamine reduced the DCF formation induced by FeSO(4) in combination with H(2)O(2). DCFH was insensitive to NO and H(2)O(2) in the cell-free system. In the presence of neutrophils, the A1242-induced luminol chemiluminescence was decreased by the superoxide dismutase inhibitor diethyldithiocarbamic acid (DDC) and the myeloperoxidase inhibitor salicylhydroxamic acid (SHA). Exposure of the neutrophils to NO, FeSO(4), or H(2)O(2) alone did not have any effect. A1242-induced lucigenin chemiluminescence in the neutrophils was increased slightly by DDC, but was not affected by SHA, NO, FeSO(4), or H(2)O(2). In conclusion, we suggest that the DCF assay is only suitable for measurements of ONOO(-), H(2)O(2) in combination with cellular peroxidases, and z.rad;OH. Luminol is sensitive towards HOCl, while lucigenin is oxidized by O(2)z.rad;(-).

Acridines↗

PubMed2003

Degenerative patterns following denervation of temporal structures in a rat model of mnemonic dysfunction.

The temporal region makes up an important substrate for mnemonic function in both humans and animals. Rats with transection of fibers connecting the temporal and entorhinal cortices display impaired visual memory. The use of hematoxylin and eosin (HE) or Nissl staining has proved insufficient for characterization of the degenerative events that take place after the lesions, and in the present study we therefore applied Fluoro-Jade dye, and zinc autometallography (AMG) in order to describe degenerative changes with AMG. We found that 14 days survival resulted in a much more severe degeneration of cell bodies in the temporal, perirhinal, entorhinal, and postrhinal cortices than was seen after 8 days survival. Corresponding degeneration of cell bodies was optimally observed 7 days following surgery with the Fluoro-Jade staining. HE staining did not reveal degenerative changes with survival times of 8 or 14 days. A substantial increase in AMG staining for zinc ions after lesion revealed an increase in zinc enriched neuronal terminals in the temporal areas and was interpreted as sprouting from local zinc enriched neuronal projections. The present data are related to the memory deficits observed in rats with similar denervations in previous studies.

Animals↗

PubMed2003

Discussion of the role of the extracellular signal-regulated kinase-phospholipase A2 pathway in production of reactive oxygen species in Alzheimer's disease.

In this paper we show that exposure of a rat brain synaptosome fraction to the amyloid beta peptide fragment betaA(25-35), but not the inverted peptide betaA(35-25), stimulated production of reactive oxygen species (ROS) in a concentration- and time-dependent manner. The ROS formation was attenuated by the tyrosine kinase inhibitor genistein, the mitogen-activated protein kinase inhibitor U0126, and the phospholipase A2 (PLA2) inhibitor 7,7-dimethyl-(5Z,8Z)-eicosadienoic acid. This strongly suggests that betaA(25-35) stimulated ROS production through an extracellular signal-regulated kinase-PLA2-dependent pathway. The interaction between these enzymes and their possible involvement in free radical formation in Alzheimer's disease are discussed.

Alzheimer Disease↗

PubMed2003

Identification of the hydroxyl radical and other reactive oxygen species in human neutrophil granulocytes exposed to a fragment of the amyloid beta peptide.

A fragment of the amyloid beta protein, betaA(25-35), was investigated for its effect on production of reactive oxygen species (ROS) in human neutrophil granulocytes. The formation and identification of ROS were examined by using a 2',7'-dichlorofluorescin (DCF) fluorescence assay, a luminol chemiluminescence assay, electron paramagnetic resonance (EPR) spectroscopy with DEPMPO as a spin trap, and hydroxylation of 4-hydroxybenzoate (4-HBA). The DCF assay showed that betaA(25-35) stimulated formation of ROS in concentration and time dependent manner. The inverted peptide, betaA(35-25), gave no response. Also, luminol-amplified chemiluminescence was stimulated by betaA(25-35). Incubation with diethyldithiocarbamate (a superoxide dimustase inhibitor) and salicylhydroxamate (SHA; a myeloperoxidase inhibitor) reduced the chemiluminescence. This indicates that hypochlorous acid (HOCl) is formed after exposure to betaA(25-35). The EPR spectra indicated a concentration dependent formation of superoxide (O2*-)- and hydroxyl (*OH)-radicals. Hydroxylation of 4-HBA to 3,4,-dihydroxybenzoate confirmed production of *OH. This response was attenuated by SHA, indicating involvement of HOCl in formation of *OH. The DCF fluorescence was inhibited with U0126 (an extracellular signal regulated protein kinase (ERK) inhibitor). Further analysis with western blot confirmed phosphorylation of ERK1/2 after exposure to betaA(25-35). The phospholipase A2 (PLA2) inhibitor 7,7-dimethyl-(5Z,8Z)-eicosadienoic acid, and diphenyleneiodonium, which inhibits the NADPH oxidase, also led to a reduction of the DCF fluorescence. The present findings indicate that betaA(25-35) stimulates the NADPH oxidase by activating the ERK pathway and PLA2. Production of O2*- can lead to HOCl and further formation of *OH, which both have a cytotxic potential.

Adult↗

PubMed2002

Effects of in vivo treatment of rats with trimethyltin chloride on respiratory properties of rat liver mitochondria.

Liver mitochondria isolated from rats treated in vivo with trimethyltin chloride show stimulation of respiration using glutamate/malate as substrate, and a transient inhibition on rates of respiration using palmitoyl-L-carnitine as substrate. This phenomenon was observed with both ADP- and FCCP-stimulated respiration. In contrast, rates of respiration by liver mitochondria isolated from rats treated in vivo with trimethyltin chloride, following prior treatment with clofibrate, were inhibited when glutamate/malate was respiratory substrates. With palmitoyl-L-carnitine no effect of trimethyltin chloride was observed. In vitro treatment of rat liver mitochondria, or of rat liver homogenates, led to the expected, powerful inhibition of respiration. The synthesis of ATP by liver mitochondria isolated from rats treated in vivo with trimethyltin chloride was not inhibited compared to mitochondria isolated from control rats. Similarly, ATP synthesis by mitochondria isolated from rats treated with clofibrate, before treatment with trimethyltin chloride, was not inhibited. We, therefore, conclude that the powerful inhibitory effects of trimethyltin found in vitro, is not expressed in vivo during the first 36 hr following administration. In vivo treatment of rats with trimethyltin chloride caused a marked increase in hepatic levels of taurine and glycine, while levels of glutathione and glutamine were diminished. This is consistent with an enhanced oxidative stress in the liver. Our findings lead to the conclusion that increased oxidative stress, rather than inhibition of the mitochondrial ATPase, is a likely major cause of the in vivo toxic effects due to trimethyltin chloride.

Adenosine Triphosphate↗

PubMed2002

Ortho-substituted polybrominated biphenyls activate respiratory burst in granulocytes from humans.

The in vivo consequences of exposure to polychlorinated biphenyls (PCB) have been reported to involve reduced phagocytic function, which could be related to increased susceptibility to infections. Though less abundant in the environment, polybrominated biphenyls (PBB) have similar toxicological properties as PCB. In this respect the effect of different PBBs on human granulocytes was elucidated. Ortho-substituted PBBs activated respiratory burst, measured by the chemiluminescence assay, and elevated intracellular calcium. The most active polybrominated congener 2,2',5-TBB increased chemiluminescence in a concentration-dependent manner, and ED(50) was approximately 10 microM. PBBs stimulated elevation of intracellular [Ca(2+)] in human granulocytes. The [Ca(2+)]i was elevated from 50 to 250 nM. The respiratory burst due to stimulation by PBBs was inhibited by U73122, ethanol (1%), wortmannin, and bisindolylmaleimide and by the elimination of extracellular calcium in the same way as shown previously for PCBs, indicating that PBB act by the same mechanisms.

Calcium↗

PubMed2002

The polychlorinated biphenyl mixture aroclor 1254 induces death of rat cerebellar granule cells: the involvement of the N-methyl-D-aspartate receptor and reactive oxygen species.

Polychlorinated biphenyls (PCBs) are widespread persistent environmental contaminants that display a complex spectrum of toxicological properties, including neurotoxicity. The present study investigates the effects of the PCB mixtures Aroclor 1242 (A1242) and Aroclor 1254 (A1254), and the PCB congeners 126 (3,3',4,4',5,-PeCB) and 153 (2,2',4,4',5,5'-HxCB) on formation of reactive oxygen species (ROS) and cell death in cultured rat cerebellar granule cells. The increase of ROS and induction of cell death were assayed using the fluorescent probe 2,7-dichlorofluorescin diacetate (DCFH-DA) and the trypan blue exclusion assay, respectively. A1242 and A1254 and PCB 153 induced a concentration-dependent increase in cell death and ROS formation. A1254 was selected for mechanistic studies. When the cerebellar granule cells were exposed to 15 microM A1254 for 12 h, 95% of the cells died. Both PCB-mediated cell death and the increase of the ROS formation were inhibited by MK-801, demonstrating the importance of the N-methyl-D-aspartate receptor. Inhibitors of nitric oxide synthase and phospholipase A2 led to a significant reduction of the DCF fluorescence and cell death. The mitochondrial permeability transition pore blocker cyclosporin A and the antioxidant vitamin E also increased survival and reduced ROS formation. The results show a connection between cell death and free radical formation.

Animals↗

PubMedDate not supplied

Short- and long-term effects of MDMA ("ecstasy") on synaptosomal and vesicular uptake of neurotransmitters in vitro and ex vivo.

3,4-Methylenedioxymethamphetamine (MDMA, "ecstasy") is a commonly abused drug which has been shown to be neurotoxic to serotonergic neurons in many species. The exact mechanism responsible for the neurotoxicity of MDMA is, however, poorly understood. In this study, the effects of MDMA on the synaptosomal and vesicular uptake of neurotransmitters were investigated. Our results show that MDMA (0.5-20 microM) reduces both synaptosomal and vesicular uptake of serotonin and dopamine in a dose dependent manner in vitro, while the uptake of glutamate and gamma-aminobutyric acid (GABA) remains unaffected. Ex vivo experiments support the importance of the monoamines, with predominant dopaminergic inhibition at short-term exposure (3 x 15 mg/kg; 2-h intervals), and exclusively serotonergic inhibition at long-term exposure (2 x 10 mg/kg per day; 4 days). This study also compares MDMA and the structurally related antidepressant paroxetine, in an attempt to reveal possible cellular mechanisms for the serotonergic toxicity of MDMA. One important difference between paroxetine and MDMA is that only MDMA has the capability of inhibiting vesicular uptake of monoamines at doses used. We suggest that inhibition of the vesicular monoamine transporter-2, and a following increase in cytoplasmatic monoamine concentrations, might be crucial for the neurotoxic effect of MDMA.

Animals↗

PubMedDate not supplied

The effect of brominated flame retardants on neurotransmitter uptake into rat brain synaptosomes and vesicles.

The environmental levels of brominated flame retardants (BFRs) are increasing, but little is known about their toxic effects. In this paper, we show that some of the most important BFRs in commercial use today, have a neurotoxicological potential. Hexabromocyclododecane (HBCD) and tetrabromobisphenol-A (TBBPA) inhibit plasma membrane uptake of the neurotransmitters dopamine, glutamate and gamma-amino-n-butyric acid (GABA) at a concentration level similar to what previously found for polychlorinated biphenyls (PCBs) and even for ecstasy. The IC(50) value for HBCD on dopamine uptake was 4 microM, and the IC(50) values for TBBPA were 9, 6 and 16 microM for dopamine, glutamate and GABA, respectively. HBCD also inhibited glutamate uptake at low concentrations, but never achieved more than 50% inhibition. The inhibition was primarily due to their effect on the membrane potential, measured by the membrane potential marker tetraphenylphosphonium bromide (TPP(+)). Other brominated flame retardants such as octaBDE and decaBDE did not have any effects on uptake. TBBPA, HBCD and even the pentabrominated diphenylether mixture (pentaBDE, DE-71, Great Lakes) also inhibited the vesicular uptake of dopamine with an IC(50) value of 3, 3 and 8 microM, respectively. The neurotoxicological consequences of these findings for environmental contaminants such as BFRs and PCBs are discussed.

Animals↗