Total body 4.5 Gy gamma irradiation-induced early delayed learning and memory dysfunction in the rat.

In an attempt to determine the consequences of total body radiation damage on learning and memory in the rat, twenty-eight male Wistar rats aged 4 months received 4.5 Gy total body gamma-irradiation (TBI) while 28 rats received sham irradiation. Sequential behavioral studies of negative reinforcement including a/ one- and b/ two-way avoidance tasks were undertaken. a/ One-way avoidance test: this test was performed before and after TBI. Prior to irradiation both groups were similar. At 20 days (D) and at 3 months post-TBI, irradiated rats had a significantly lower percentage of avoidance than controls but no statistical difference was found at 5 months post-TBI. b/ Two-way avoidance test: this test was performed only after TBI. At days 21, 22, 23, 24, (leaming) and at 4 or 6 months (recalls) post-TBI the mean percentage of avoidance was significantly lower in irradiated than in control rats. This study demonstrates that total-body exposure to 4.5 Gy gamma-irradiation induces behavioral dysfunction affecting learning and transitorily memory. These results suggest that a relatively low dose of total body irradiation can induce neurological complications, which persist 4-6 months later.

Low dose of the gamma acute radiation syndrome (1.5 Gy) does not significantly alter either cognitive behavior or dopaminergic and serotoninergic metabolism.

The aim of this study was to evaluate the early-delayed effects of a low dose of the gamma acute radiation syndrome (1.5 Gy) on memory and on dopaminergic and serotoninergic metabolism in Swiss albino CD1 mice, of various ages (6, 10 and 20 weeks). At different times after irradiation (from 24 hr to three months), the mice were trained in a single-trial passive avoidance task and tested for retention either 24 hr or 5 days later. Their performance was compared to that of mice that were sham-irradiated. At the end of the behavioral test (days 3, 9, 30 and 93), the concentrations of dopamine (DA) and serotonin (5HT) and their metabolites were determined in hippocampus, anterior cortex and striatum of mice irradiated at the age of six weeks. No significant behavioral effect was observed whichever the age of the animals or the delay of observation. On the contrary at the moderate dose of 4.5 Gy we observed a significant memory deficit 9 days after the exposure. Considering the neurochemical study, in the striatum or in the frontal cortex, no significant modification was observed whichever the delay or the molecule. In the hippocampus slight modifications were noted: an increase (+144%, p = 0.002) in DA level on day 3 after exposure, and a decrease (-27%, p = 0.028) of 5HT level on day 30 post-irradiation. These modifications were only transient and not associated to modifications of the catabolites. This study demonstrates that total-body exposure to gamma radiation at low dose seems to induce only slight effects on the central nervous system.

Prolonged effects of acute gamma irradiation on acetylcholine-induced potassium currents in human umbilical vein endothelial cells.

Bourlier, V., Diserbo, M., Gourmelon, P. and Verdetti, J. Prolonged Effects of Acute Gamma Irradiation on Acetylcholine-Induced Potassium Currents in Human Umbilical Vein Endothelial Cells. Radiat. Res. 155, 748-752 (2001). We have recently reported an acute effect of gamma irradiation (15 Gy, 1 Gy/min) on acetylcholine-mediated endothelium-dependent relaxation in rat aortic rings. Given the importance of permeability to K+ to endothelium-dependent relaxation, we have evaluated the effect of the same radiation on K+ currents in human endothelial cells in culture using the patch-clamp technique in the whole-cell recording configuration. Our results indicate that, in resting cells, gamma irradiation has no effect on endothelial permeability to K+. However, irradiation during stimulation of endothelial cells with acetylcholine reduces the sustained increase in permeability to K+ observed in the acetylcholine-stimulated, nonirradiated cells. Additional experiments using K+ channel inhibitors (TEA, charybdotoxin, apamin) suggest that irradiation may in part decrease the prolonged activation of Ca2+-activated K+ channels by acetylcholine. Taken together with our previous finding that irradiation inhibits the acute relaxing effects of acetylcholine, these results show that gamma irradiation also affects the delayed effects of acetylcholine on permeability to K+.

Utility of the wavelet transform to analyze the stationarity of single ionic channel recordings.

Wavelet transform, a time-scale analysis, is presented as a new tool to analyze single-channel recordings. This method makes it possible to verify the stationarity, to identify episodes of change in the kinetic channel behavior (burst, flickering, cooperativity) or episodes of noise, and to localize stationary segments in long single-channel current recordings. It can help the conventional analysis of the kinetic behavior of ionic channels leading to better understand the gating mechanism.

Early effects of acute gamma-radiation on vascular arterial tone.

1. To determine the acute effects of irradiation on the functionality of vessel, rat aortic rings were mounted in an organ bath for isometric tension measurements and irradiated (60Co, 1 Gy min(-1), 15 min). 2. Irradiation, which is without effect on non-contracted or endothelium-denuded vessels, led to an immediate and reversible increase in vascular tone on (-)-phenylephrine (1 microM)-precontracted aortic rings. The tension reached a plateau about 5 min after the beginning of irradiation. 3. The maximal radiation-induced contraction occurred on aortic rings relaxed by acetylcholine (ACh) (1 microM). In this condition, the addition of catalase (1000 u ml(-1)), which reduces hydrogen peroxide, and DMSO (0.1% v/v), which scavenges hydroxyl radical, had no influence on tension level while superoxide dismutase (SOD) (100 u ml(-1)), a superoxide anion scavenger, reduced the observed contraction. A similar result was obtained in the presence of indomethacin (10 microM), a cyclo-oxygenase blocker. 4. Pretreatment of rings with the nitric oxide synthase inhibitor, N(omega)-nitro-L-arginine methyl ester (L-NAME) (10-100 microM) inhibited the radiation-induced contraction. 5. This effect was dose rate-dependent and even occurred for a very low dose rate (0.06 Gy min(-1)). 6. The present results indicate that gamma-radiation induces an instantaneous vascular tone increase that is endothelium and dose rate-dependent. This effect is (i) maximal when nitric oxide (NO) is produced, (ii) greatly reduced by SOD and (iii) inhibited by L-NAME, suggesting a major involvement of complexes between NO and superoxide anion.

Activation of large conductance Ca(2+)-activated K+ channels in N1E-115 neuroblastoma cells by platelet-activating factor.

Using patch-clamp single channel recording techniques, we reported a Platelet-Activating Factor "PAF"-induced activation of large conductance Ca(2+)-activated K+ "BK(Ca)" channels in N1E-115 cells. This activation was only observed in cell-attached configuration and was blocked by the PAF antagonist BN50739 or removal of calcium from the bath. Nanomolar concentration of PAF produced a transient hyperpolarization observed in whole-cell current clamp configuration which was blocked by the bath application of BN50739 or iberiotoxin. Our results suggest that the PAF-induced hyperpolarization is mediated by an activation of BK(Ca) channels coupled to specific PAF receptors. This coupling is not direct and results from the PAF-induced elevation in cytosolic free Ca2+ concentrations that we have previously described in N1E-115 cells (Cell Calcium 1995, 17, 442-452).

Stimulation of platelet-activating factor (PAF) receptors increases inositol phosphate production and cytosolic free Ca2+ concentrations in N1E-115 neuroblastoma cells.

Platelet-activating factor (1-O-alkyl-2-acetyl-sn-glycero-3-phosphorylcholine, PAF) has recently been recognized as an important mediator in the pathophysiology of brain injury. This study demonstrates that, in suspended populations of N1E-115 cells loaded with Indo-1, biologically relevant concentrations of PAF produce a rapid and transient elevation in cytosolic free calcium concentration ([Ca2+]i). Moreover, nanomolar concentrations of PAF increase [3H]-inositol phosphate production. Using lyso-PAF and the specific PAF-receptor antagonists BN52021 and BN50739, we show that these effects were mediated by stimulation of PAF receptors. Experiments performed in Ca2+ free medium show that PAF-induced [Ca2+]i increase is the result of an influx of Ca2+ and of the release of intracellular Ca2+ stores. Studies of Mn2+ influx argue in favour of additional pathways for the PAF-induced Ca2+ influx other than the pathway for the thapsigargin-induced Ca2+ influx. Using the whole-cell voltage-clamp technique, we observe that PAF induces an increase of Ltype Ca2+ current. However, the effects of La3+, nifedipine and KCl-induced depolarization on the PAF-induced [Ca2+]i increase suggest a minor participation of these voltage-gated Ca2+ channels in the response to PAF. Altogether the results point to the existence of a PAF-induced Ca2+ influx through receptor-operated Ca2+ permeant channels.

Presence of specific platelet-activating factor binding-sites in neuroblastoma N1E-115 cells.

In this study we reported evidence for the existence of specific binding sites for platelet-activating factor (PAF) in neuroblastoma N1E-115 cells. The specific [3H]PAF binding reached a steady state level within 60 min at 25 degrees C. Scatchard analysis of the specific [3H]PAF binding revealed the presence of two apparent populations of binding sites. The high-affinity binding site possessed a Kd1 of 2.5 +/- 0.6 pM and Bmax1 = 57.3 +/- 20.0 fmol/mg protein. The low-affinity binding site possessed a Kd2 = 3.2 +/- 1.0 nM and Bmax2 = 4.4 +/- 2.1 pmol/mg protein. Furthermore, the total [3H]PAF binding was partially displaced by unlabelled PAF, PAF antagonists BN 52021 and BN 50730 in a dose-dependent manner. This study confirms the presence of specific PAF receptors in neuronal cells.

Biophysical and pharmacological properties of large conductance Ca(2+)-activated K+ channels in N1E-115 cells.

The large conductance Ca(2+)-activated K+ channels in differentiated mouse neuroblastoma N1E-115 cells have been studied using patch-clamp single-channel current recording in excised membrane patches. These channels displayed a unitary conductance of 200 pS under symmetrical K+ concentrations. Effects of blockade by TEA+, Cs+ and Ba2+ were different and argued for distinct action mechanisms. The open probability of these channels increased with increasing internal calcium and membrane potential. Maximum sensitivity of these channels ranged over physiological variations of internal calcium at membrane potentials close to zero, suggesting a physiological role for these channels in regulating the membrane potential and Ca2+ influx through voltage-dependent Ca2+ channels.