Effect of chronic gamma-irradiation and beta-carotene on lipid metabolism in presynaptic membranes in rat brain.

Lysophosphatidylcholine is involved in radiation-induced modulation of presynaptic membranes in rat brain. High sensitivity of the cortical integrative functions to chronic low-dose gamma-irradiation is demonstrated. beta-Carotene produced a protective effect during chronic irradiation.

[Behavior reactions and lipids of brain synaptic membranes of rats under chronic exposure to gamma irradiation]. / Povedencheskie reaktsii i lipidy sinapticheskikh membran golovnogo mozga u krys pri khronicheskom vozdeistvii gamma-izlucheniia.

The effects of low level chronic ionising irradiation (12.9 cGy/day on the sensory attention to the stimuli of different modalities (somatosensory, visual, odor) of Wistar rats were studied. Analysis of animals behaviour was made after they had received the different doses of irradiation: 4, 6, 8, 10, 15 and 20 Gy. It was founded, that the attention and exploratory activity of rats is significantly decreased up to 20-30% after 4-6 Gy. The irradiation doses 8 Gy did not change animal behaviour as compared to control animals, but doses 10, 15 and 20 Gy decreased the exploratory activity as well as sensory attention of rats to 3-5-times as compared to previous dose. Such a wave-like way of behaviour reflects the functioning of an adaptive mechanism. Biochemical data indicated that after 5 months of the irradiation (dose 20 Gy) the level of phospholipids, lysophosphatidylcholine, phosphatidylethanolamine, phosphatidylcholine, cholesterol were decreased.

[The effect of antioxidants on functional brain disorders caused by low-intensity ionizing radiation]. / Issledovanie vliianiia antioksidantov na narusheniia funktsional'noi aktivnosti mozga, vyzvannye ioniziruiushchim izlucheniem maloi moshchnosti.

The effects of daily intraperitoneal injections of alpha-tocopherol (30 mg/kg per day) and synthetic antioxidant IHFAN-30 (30 mg/day) in rats were compared during low-level ionizing radiation (10 days, dose rate 5 mGy/h, total dose 1.2 Gy). There were analysed: (1) amplitude of population spike of hippocampal slices; (2) endogenous phosphorylation in vitro of hippocampal synaptic proteins in the presence of cAMP; (3) formation, manifestation and reduction of food-procuring reflex. The findings showed that antioxidants made some correction of the functional state of hippocampal slices and cAMP-dependent phosphorylation system activity in brain cells from irradiated animals. No influence on training and memory functions was detected.

Effect of gamma-irradiated carbohydrates on isolated synaptic membranes.

The effect of gamma-irradiated solutions of carbohydrates, mainly glucose, upon Na+, K(+)-ATPase and lipid peroxidation in rat brain synaptosomal membranes was studied. The membrane damage by irradiated glucose was enhanced in the presence of Fe2+ and was diminished when a free-radical scavenger (BHT) or metal chelators (EDTA, EGTA) were present. It is suggested that a key element in the free-radical membrane damage by irradiated carbohydrates is an Fe(2+)-complex of some species of the radiolysis products. Participation of radiotoxins of carbohydrate origin in radiobiological effects is discussed.

[Action of microwaves with different modulation frequencies and exposure times on GABA receptor concentration in the cerebral cortex of rats]. / Deistvie mikrovoln s raznoi chastotoi moduliatsii i vremenem ékspozitsii na kontsentratsiiu retseptorov GAMK v kore mozga krys.

The effect of 800 mHz microwaves of 0, 3, 5, 7, 16, and 30 Hz modulation on GABA receptor concentration in rat brain cortex has been investigated. Irradiation of the whole body at a modulation frequency of 16 Hz readily decreases the GABA receptor concentration. Irradiation at other modulation frequencies is ineffective. Irradiation of the whole body modulated at 16 Hz with various exposure times (5, 15 and 60 min) has revealed the highest effect at 5 min, while at a longer exposure the effect decreases.

Molecular target size of the vanilloid (capsaicin) receptor in pig dorsal root ganglia.

The size of the vanilloid receptor was examined by high-energy radiation inactivation analysis of the binding of [3H]resiniferatoxin to pig dorsal root ganglion membranes; it was found to be 270 +/- 25 kDa. This value most likely represents the size of a receptor complex rather than of an individual subunit. Other ligand-gated cation channel complexes have reported molecular weights in this range, e.g. 300 kDa for the acetylcholine receptor.

[Functional changes in the synaptic membrane during in vitro interaction with bilirubin and serum albumin]. / Funktsional'nye izmeneniia v sinaptosomal'noi membrane pri vzaimodeistvii s bilirubinom i syvorotochnym al'buminom in vitro.

A decrease in total activity of ATPase and in specific activity of Na+,K+-ATPase as well as less rapid and distinct decrease in specific activity of acetylcholinesterase were observed in synaptosomal membrane after binding of bilirubin in vitro. Irradiation with blue light of the bilirubin-containing membrane particles caused a more distinct decrease in activity of these enzymes. Blood serum albumin, added to the suspension of bilirubin-containing particles of synaptosomal membrane, affected significantly the alterations in activity of membrane-bound enzymes caused by the irradiation. Character of the effect depended on pH of the medium and presence of organophilic ligands in blood serum albumin molecule.

[The effect of ionizing radiation on the binding of muscimol by synaptic membranes in the rat brain]. / Deistvie ioniziruiushchei radiatsii na sviazyvanie mustsimola sinapticheskimi membranami mozga krysy.

A study was made of the effect of gamma-radiation on binding of muscimol, a GABA agonist, by synaptic membranes of rat brain cortex. Exposure to 2 Gy radiation was shown to reduce [3H]-muscimol binding to membranes.

[ATP-dependent Ca2+-transporting system in the rat brain during an early period of acute radiation injury]. / ATF-zavisimaia Ca2+transportiruiushchaia sistema mozga krys na rannem étape ostrogo luchevogo vozdeistviia.

The rate of Mg2+, Ca2+-ATPase reaction and ATP-dependent Ca2+ accumulation in a preparation of plasma membranes from brain synaptosomes increases 60 min following whole-body X-irradiation of rats with a dose of 0.21 C/kg, a calcium sensitivity of both processes being increased. A unidirectional change in their kinetics indicates the early radiosensitivity of Ca2+ transfer systems in the brain synaptosome membranes. There is an increase in the availability of SH-groups of membrane preparation proteins for SH-reagents and in the sensitivity of Mg2+, Ca2+-ATPase reaction and ATP-dependent Ca2+ accumulation to trifluoperazine, a calmodulin inhibitor. Both processes lose their ability to be activated by exogenous calmodulin. It is suggested that at an early stage of radiation affection, a change occurs in the molecular organization of the ATPase-calmodulin membrane complex in plasma membranes of rat brain synaptosomes.

Target size analysis of opioid receptors. No difference between receptor types, but discrimination between two receptor states.

Target size analysis of opioid receptor is complicated by the presence of multi-exponential inactivation curves. Irradiation of intact frozen tissue proved essential to eliminate such artifacts, due to indirect irradiation effects. Upon irradiation condition, opioid binding activity was inactivated in a single mono-exponential manner. Identical inactivation curves were obtained for mu, delta and kappa binding activities in brain membranes from rat, guinea-pig and frog and in NG 108-15 cells: the molecular mass obtained was 98 +/- 2 kDa. However, when opioid binding was assayed in the presence of Na+, Mg2+ and GTP, the molecular mass was found to be only 56 +/- 4.4 kDa. We suggest that the opioid recognition site comprises a unit of 56 kDa and that in the absence of Na+, Mg2+ and GTP an additional membrane component of 40-44 kDa is necessary for high-affinity opioid binding.

Molecular properties of the apamin-binding component of the Ca2+-dependent K+ channel. Radiation-inactivation, affinity labelling and solubilization.

Radiation-inactivation was used to assess the functional size of the apamin-binding component of the Ca2+-dependent K+ channel. The amount of specific binding of 125I-apamin to receptors in synaptic membranes of rat cortex decayed exponentially with increasing doses of ionizing radiation and target size analysis was consistent with a relative molecular mass of 250 000 +/- 20 000 for the 125I-apamin receptor. Analysis on sodium dodecyl sulfate gels following covalent cross-linking of 125I-apamin to its receptor in a synaptosomal membrane preparation from rat cortex revealed a single labelled polypeptide chain of Mr = 33 000 +/- 2000 in the presence of protease inhibitors. Our results suggest that the Ca2+-dependent K+ channel from rat cortex is an oligomeric structure of Mr = 250 000 +/- 20 000 containing an apamin-binding subunit of Mr = 33 000 +/- 2000. The apamin-binding component of the Ca2+-dependent K+ channel from rat synaptosomes was solubilized using detergents such as sodium cholate or 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate. Phospholipids did not increase the stability of the apamin-binding component during the solubilization. Binding of apamin to its solubilized receptor is reversible and saturable. The dissociation constant of the apamin-receptor complex is 40-150 pM, the rates constants of association and dissociation being 3.2 X 10(6) M-1s-1 and 1.4 X 10(-4)s-1 respectively. These binding characteristics are similar to those found for the membrane-bound apamin receptor.

[Effect of ultra-high frequency electromagnetic waves on a postsynaptic membrane model]. / Deistvie sverkhvysokochastotnykh élektromagnitnykh voln na model' postsinapticheskoi membrany.

It was shown that high-frequency electromagnetic field increases the conductivity of ionic channels formed by the synaptic membrane fragments, which bind glutamate, in a bilayer lipid membrane. Heating of the whole electrolyte in a cell, under the effect of the electromagnetic field applied, was minor to be responsible for the effects observed.

Photoaffinity-labelling of the glycine receptor of rat spinal cord.

The irreversible incorporation upon ultraviolet illumination of the glycine receptor antagonist, [3H]strychnine, into synaptic membrane fractions of rat spinal cord has been investigated. The specificity of this photoaffinity-labelling reaction for the glycine receptor was demonstrated by the following results: (a) the Kd value (9.7 nM) of the glycine-displaceable irreversible incorporation of [3H]strychnine was similar to the previously reported Kd of [3H]strychnine binding to the glycine receptor; (b) pre-illumination of the membranes with unlabelled strychnine led to a corresponding reduction in the number, but not the affinity, of reversible glycine-displaceable [3H]strychnine binding sites; (c) the ultraviolet light-induced incorporation into the membranes of [3H]strychnine was inhibited by different glycine receptor agonists; other neurotransmitter substances had little or no effect. Also, [3H]strychnine alone was shown to be stable upon illumination with ultraviolet light; this suggests that photocrosslinking of [3H]strychnine may require energy transfer from specific groups of its high-affinity receptor binding site. Upon sodium dodecyl sulphate/polyacrylamide gel electrophoresis a single labelled polypeptide with a relative molecular mass of 48000 was revealed from spinal cord membranes photoaffinity-labelled with [3H]strychnine. Spinal cord membranes photoaffinity-labelled with the gamma-aminobutyric acid receptor ligand [3H]flunitrazepam, however, gave a single polypeptide with a relative molecular mass of 5- 0000. Treatment of membranes, labelled with [3H]strychnine, by endoglycosidase H did not alter the relative molecular mass of the 48000-Mr labelled polypeptide. Trypsin treatment, on the other hand, successively produced major fragments of relative molecular masses of 42000 and 37000. Also, even after extensive treatment with trypsin or chymotrypsin, greater than or equal to 90% of the radioactivity incorporated into the labelled membranes remained membrane-associated. It is concluded that the strychnine binding site of the glycine receptor is located on a protease-inaccessible, i.e. probably hydrophobic domain of the 48000-Mr subunit.

Determination of the molecular size of the nitrendipine-sensitive Ca2+ channel by radiation inactivation.

The molecular size of the [3H] nitrendipine receptor of transverse tubules prepared from rabbit skeletal muscle and from rat cortex synaptic membranes have been investigated. Radiation inactivation of the specific binding of [3H] nitrendipine was consistent with Mr equals 210 000 +/- 20,000 for the receptor in each membrane preparation indicating a common size for the [3H] nitrendipine receptor.