Effect of [His7]-corazonin on the number of antennal sensilla in Locusta migratoria

Abstract.  Certain types of antennal sensilla are known to be more abundant in solitarious individuals than in gregarious ones in the migratory locust, Locusta migratoria . We tested the hypothesis that injection of a neurohormone, [His⁷]-corazonin, into isolated-reared nymphs of this species mimics the effect of crowding on the frequencies of various types of antennal sensilla. One nmol of the hormone was injected into nymphs on two occasions, on the third days of the second and third stadia, respectively. Upon adult emergence, the numbers of different types of sensilla on the eighth antennal segment were compared with those of oil-injected controls. [His⁷]-corazonin did not influence the numbers of basiconic sensilla type A, basiconic sensilla type B and trichoid sensilla significantly compared to oil-injected controls. However, the number of coeloconic sensilla was reduced significantly by the hormone injections. Because the length of the antennal segment was not affected by the hormone injection, it appears that the hormone influenced the development of coeloconic sensilla. The results support the hypothesis tested and are consistent with the idea that [His⁷]-corazonin plays an important role in the control of phase polymorphism in L. migratoria .     

Albino corpus cardiacum extracts induce morphometric gregarization in isolated albino locusts, Locusta migratoria, that are deficient in corazonin

Abstract.  The neuropeptide [His⁷]-corazonin, present in the central nervous system and corpus cardiacum, is known to mimic a 'gregarizing' effect on phase-related morphometric ratios (hind femur length/maximum head width and fore wing length/hind femur length) when injected into locusts reared in isolation. However, an albino strain is known to exhibit phase-specific changes in these ratios in response to rearing density, although it is deficient in [His⁷]-corazonin. To examine whether there is a second factor responsible for this phenomenon, perhaps a corazonin-like factor that has lost its dark-colour inducing activity, methanol extracts of corpora cardiaca taken from crowd-reared albino nymphs of Locusta migratoria are injected into isolated-reared second-stadium albino nymphs and reared to adults in isolation. The hind femur length/maximum head width and fore wing length/hind femur length ratios are significantly different from those of control oil-injected counterparts, and shift significantly towards the values typical for crowd-reared gregarized individuals. The results indicate that the corpora cardiaca contain a factor similar to [His⁷]-corazonin, although it has no dark-colour inducing activity.     

Behavioural differences in Locusta migratoria associated with albinism and their relation to [His7]-corazonin

Abstract. The neuropeptide [His⁷]-corazonin induces black colouration in locusts, a feature typically occurring in gregarious animals. To date, the function of this neuropeptide in relation to phase transition has not been fully clarified. Unanswered questions on its involvement in behavioural and morphometrical phaseshifting and possible temperature-induced regulation still remain. In the African locust, Locusta migratoria migratorioides (Reiche and Fairmaire) (Orthoptera: Acrididae), the gregarious form possesses mainly a brown colouration with typical intense black pigmentation. To find a possible behavioural function, an albino L. migratoria mutant, deficient in [His⁷]-corazonin and thus lacking the typical gregarious colour, was used. Between these two phenotypes, nonphase specific behavioural differences in a phase-dependent behavioural set-up were found. Additionally, a wild-type of the Okinawa strain was screened behaviourally and located in comparison to the other groups. Treatment of albinos with corazonin induced normal phenotype behaviour. The degree of interference of corazonin in relation to these findings and a possible effect at the level of visual perception is discussed.     

Age-dependent response of adults of a homochrome grasshopper, Oedipoda miniata, to the dark-colour-inducing neurohormone (DCIN) of locusts

Abstract The effect of the dark-colour-inducing neurohormone (DCIN = [His⁷]-corazonin) of locusts was investigated in field-collected young and old adults of the grasshopper, Oedipoda miniata (Pallas) (Orthoptera: Acrididae). This species shows homochromy, but neither green-brown, nor phase-dependent colour polymorphism. By injecting graded doses of synthetic DCIN in 2 µL of olive oil, young adults were tested within a week of their last moult, and old adults 3.5 months later, a few weeks before natural termination of their reproductive summer diapause. Colour changes were followed for 28 days after injection. Darkening of the young adults was considerable, but their response to DCIN is more moderate than that of conspecific nymphs, by exhibiting a higher threshold, slower response and weaker maximal response. Old adults also show a clear effect, but their response is even slower and less marked than the response of the young adults. It is concluded that the response to DCIN decreases from nymphs to young adults and it is further decreasing with ageing of the adults.     

The dark-colour-inducing neurohormone of locusts: strain-dependent and phase-independent effects on adults of the migratory locust, Locusta migratoria

Abstract.  An albino strain that had originated from Okinawa, Japan, and a normally coloured strain that had originated from West Africa, were used to study the darkening response to injection of graded doses of dark-colour-inducing neurohormone of locusts (DCIN) ([His⁷]-corazonin) of gregarious and solitarious adults of the migratory locust, Locusta migratoria (L.). By repeated crossings, congenic albinos and normal phenotypes were obtained, both with a 99.6% West African genome, and their darkening response was compared with the original Okinawa and West African strains. Within each of these four strains, no difference was found in DCIN-induced darkening between gregarious and solitarious adults despite previous publications in the literature claiming an absence of 'fire-darkening' in gregarious adults of this species. Okinawa albino adults showed a markedly higher darkening response than the other three strains, including albinos with a 99.6% West African genome. This finding demonstrates that the differential darkening response of the Okinawa albinos is caused not by albinism, but by the geographical origin (Okinawa) of the strain. This is the first report of geographical-strain-dependent differences in the response of an insect to a neurohormone. The darkening response of adults reached a maximum on day 10 after injection; subsequently, the dark colour faded slowly. Adults injected 1 day after their moult showed a greater darkening response than those injected after 14 or 28 days.     

Endocrine mechanisms controlling body-color polymorphism in locusts.

The present article reviews recent published and unpublished findings on the hormonal mechanisms for the control of body-color polymorphism in locusts. Emphasis is placed on the dark color-inducing factors and their role in the induction of various types of body coloration observed under different environmental conditions. Implantation of corpora cardiaca (CC) taken from normal nymphs of Locusta migratoria induced dark color in nymphs of an albino strain. Using the albino strain for the bioassay, a neuropeptide, [His7]-corazonin, was identified as a dark color-inducing factor for L. migratoria and Schistocerca gregaria. In the former, depending upon the dose and timing of the injection, this peptide and juvenile hormone developed various body colors looking like those found in nature. The body coloration characteristic of gregarious forms was also induced in isolated albino nymphs and field-collected solitary nymphs. In S. gregaria, on the other hand, the peptide induced black patterns, but the orange or yellow background color observed in gregarious forms was not induced when the peptide was injected into solitary individuals. [His7]-corazonin also induced darkening in other grasshoppers and locusts, but not in katydids. Albino L. migratoria developed dark color when implanted with brains or CC taken from other insects belonging to 10 major insect orders, but not with those from Coleoptera. [His7]-corazonin or a similar compound is widespread among insects and plays a pivotal role in controlling body color in some species and presumably other physiological roles in other species. Arch.     

Purification and biochemical characterization of a membrane-bound [NiFe]-hydrogenase from a hydrogen-oxidizing, lithotrophic bacterium, Hydrogenophaga sp. AH-24

Membrane-bound [NiFe]-hydrogenase from Hydrogenophaga sp. AH-24 was purified to homogeneity. The molecular weight was estimated as 100±10 kDa, consisting of two different subunits (62 and 37 kDa). The optimal pH values for H₂ oxidation and evolution were 8.0 and 4.0, respectively, and the activity ratio (H₂ oxidation/H₂ evolution) was 1.61 × 10² at pH 7.0. The optimal temperature was 75 °C. The enzyme was quite stable under air atmosphere (the half-life of activity was c . 48 h at 4 °C), which should be important to function in the aerobic habitat of the strain. The enzyme showed high thermal stability under anaerobic conditions, which retained full activity for over 5 h at 50 °C. The activity increased up to 2.5-fold during incubation at 50 °C under H₂. Using methylene blue as an electron acceptor, the kinetic constants of the purified membrane-bound homogenase (MBH) were V _max=336 U mg⁻¹, k _cat=560 s⁻¹, and k _cat/ K _m=2.24 × 10⁷ M⁻¹ s⁻¹. The MBH exhibited prominent electron paramagnetic resonance signals originating from [3Fe–4S]⁺ and [4Fe–4S]⁺ clusters. On the other hand, signals originating from Ni of the active center were very weak, as observed in other oxygen-stable hydrogenases from aerobic H₂-oxidizing bacteria. This is the first report of catalytic and biochemical characterization of the respiratory MBH from Hydrogenophaga .     

Hormonal control of body-color polymorphism in Locusta migratoria: interaction between

The effects of injection of [His(7)]-corazonin and juvenile hormone (JH) III on the body color in L. migratoria were investigated using albino and normal (pigmented) nymphs. Most albino nymphs turned green in the fourth instar if injected with JH III during the last 2 days of the previous instar. When albino third instar nymphs injected with 10 pmol of [His(7)]-corazonin on different days were treated with 100 μg of JH III on day 3.5, they developed various body colors in the following nymphal instar: those injected with [His(7)]-corazonin during the first 2 days developed very dark green or black color, whereas some of those injected after this period turned green and their legs and ventral side of the body were variously pigmented, the coloration being similar to green solitary individuals often found in the field. Field-collected brown solitary nymphs injected with 1 nmol of [His(7)]-corazonin and kept individually, turned reddish without any black spots in the following nymphal instar when the ecdysis occurred within 1 day after injection. Injection of [His(7)]-corazonin 2 days before the following ecdysis induced black patterns on an orange background color, the coloration characteristic of gregarious forms. Similar injections into field-collected green solitary nymphs also induced black patterns but the rest of their body remained green. These results may indicate that the temporal changes in the hemolymph titers of [His(7)]-corazonin and JH play an important role in the control of body-color polymorphism in this locust.     

Cations Affect [3H]Mazindol and [3H]WIN 35,428 Binding to the Human Dopamine Transporter in a Similar Fashion

Abstract: The present study addresses the possibility that there are different cocaine-related and mazindol-related binding domains on the dopamine transporter (DAT) that show differential sensitivity to cations. The effects of Zn²⁺, Mg²⁺, Hg²⁺, Li⁺, K⁺, and Na⁺ were assessed on the binding of [³H]mazindol and [³H]WIN 35,428 to the human (h) DAT expressed in C6 glioma cells under identical conditions for intact cell and membrane assays. The latter were performed at both 0 and 21°C. Zn²⁺ (30–100 µ M ) stimulated binding of both radioligands to membranes, with a relatively smaller effect for [³H]mazindol; Mg²⁺ (0.1–100 µ M ) had no effect; Hg²⁺ at ∼3 µ M stimulated binding to membranes, with a relatively smaller effect for [³H]mazindol than [³H]WIN 35,428 at 0°C, and at 30–100 µ M inhibited both intact cell and membrane binding; Li⁺ and K⁺ substitution (30–100 m M ) inhibited binding to membranes more severely than to intact cells; and Na⁺ substitution was strongly stimulatory. With only a few exceptions, the patterns of ion effects were remarkably similar for both radioligands at both 0 and 21°C, suggesting the involvement of common binding domains on the hDAT impacted similarly by cations. Therefore, if there are different binding domains for WIN 35,428 and mazindol, these are not affected differentially by the cations studied in the present experiments, except for the stimulatory effect of Zn²⁺ at 0 and 21°C and Hg²⁺ at 0°C.     

The role of

The effect of [His(7)]-corazonin on the body color in Locusta migratoria was examined by varying the injected dose and the time of injection in both an albino and a normal (pigmented) strain. Albino nymphs injected with a high dose (100pmol) of [His(7)]-corazonin at the beginning of the third instar turned completely black in the following instar, whereas those injected with the same dose in the middle of the instar developed black patterns with an orange background color, the body coloration characteristic of normal gregarious (crowded) individuals. Injection at the end of the third instar induced a reddish color with few black spots. Irrespective of the time of injection of the peptide, most of these individuals became completely black after ecdysis to the fifth instar. A similar result was obtained with a lower dose (1pmol), although the color expressed was lighter. In the normal strain, injection of 1nmol or 100pmol into crowded third instar nymphs also caused most of them to become completely black in the fourth and fifth instars, but a lower dose apparently had no influence. These results suggest that the temporal changes in hemolymph titer of [His(7)]-corazonin are important in the expression of body color in L. migratoria.     

Stem wood properties of mature Norway spruce after 3 years of continuous exposure to elevated [CO2] and temperature

The objective of the study was to investigate the interactive effects of elevated atmospheric carbon dioxide concentration, [CO₂], and temperature on the wood properties of mature field-grown Norway spruce ( Picea abies (L.) Karst.) trees. Material for the study was obtained from an experiment in Flakaliden, northern Sweden, where trees were grown for 3 years in whole-tree chambers at ambient (365 μmol mol⁻¹) or elevated [CO₂] (700 μmol mol⁻¹) and ambient or elevated air temperature (ambient +5.6 °C in winter and ambient +2.8 °C in summer). Elevated temperature affected both wood chemical composition and structure, but had no effect on stem radial growth. Elevated temperature decreased the concentrations of acetone-soluble extractives and soluble sugars, while mean and earlywood (EW) cell wall thickness and wood density were increased. Elevated [CO₂] had no effect on stem wood chemistry or radial growth. In wood structure, elevated [CO₂] decreased EW cell wall thickness and increased tracheid radial diameter in latewood (LW). Some significant interactions between elevated [CO₂] and temperature were found in the anatomical and physical properties of stem wood (e.g. microfibril angle, and LW cell wall thickness and density). Our results show that the wood material properties of mature Norway spruce were altered under exposure to elevated [CO₂] and temperature, although stem radial growth was not affected by the treatments.     

Energy balance and temperature relations of Azorella compacta, a high-elevation cushion plant of the central Andes

The environmental relationships and ecophysiology of Azorella compacta, a giant cushion plant, were investigated in Parque Nacional Lauca, Chile (18°10'–18°25' S and 69°16' W, 4400 m asl). The diurnal temperature range can reach 42 °C on some days of the year. The surface temperature of A. compacta was 13 °C below that of the air temperature of −7 °C at dawn, but from midmorning to late afternoon, the plant surface temperature remained within a few degrees of the air temperature. Soil surface temperatures did not differ between north- and south-facing slopes, but a model showed an increase in radiation reception by north-facing slopes throughout most of the year. Gas exchange measurements of A. compacta measured at the onset of the wet season ranged from −0.6662 to 11.4 μmol·m⁻²·s⁻¹, and maximum stomatal conductance (Gs) was 410 mmol·m⁻²·s⁻¹. The estimated light compensation point was 89 μmol·m⁻²·s⁻¹ and estimated light saturation occurred at about 1280 μmol·m⁻²·s⁻¹. Diurnal water potential measurements for A. compacta ranged from −1.67 to −2.65 MPa. This is one of the first ecophysiological studies of a tropical alpine cushion plant.     

Efficacy of Beauveria bassiana (Bals.) Vuill. against the spruce bark beetle, Ips typographus L., in the laboratory under various conditions

Abstract:  In laboratory bioassays, the efficacy of the entomopathogenic fungus Beauveria bassiana against the spruce bark beetle, Ips typographus , was tested under various conditions. Four of the tested isolates and the commercial product Boverol^® caused 99–100% mortality when tested at a concentration of 1.0 × 10⁷ conidia/ml at 25°C. Using B. bassiana isolate 138 at a concentration of 1.0 × 10⁶, the median survival time (MST) was 6.1 d and significantly longer compared with the MST of 4.2 and 4.0 d at 1.0 × 10⁷ and 1.0 × 10⁸ conidia/ml, respectively. In the next experiment, the beetles were maintained on spruce bark, filter paper or artificial diet during the bioassay with Boverol^®, and significant differences in the MST of 3.6, 2.5 and 5.3 d, respectively, were noticed. The experiment with Boverol^® at different temperatures showed that the beetles lived significantly longer at 15°C (MST 8.7 d) than at 20, 25, 30 and 35°C. At 25°C, the beetles died most rapidly (MST 3.5 d). At different relative humidities (RH) of 40, 70 and 100%, nearly all beetles were dead after treatment with a suspension of Boverol^® at 1.0 × 10⁷ conidia/ml. At 40% RH, 49% of the untreated beetles died after 7 d. The best effects were achieved with the following bioassay: beetles were fed for three days on artificial diet, then dipped into a solution of 1.0 × 10⁷ conidia/ml and transferred on a piece of spruce bark in Petri dishes at 25°C and 70% RH.     

Intergenerational acclimation in aphid overwintering

Abstract.  1. When first instar nymphs and adults of the grain aphid Sitobion avenae (Fabricius) (Hemiptera: Aphidiae) were maintained in long-term cultures (>6 months) at 20 °C and 10 °C, the LT₅₀ decreased from −8 and −8.8 °C to −16.0 and −13.5 °C, respectively.
2. When aphids from the 20 °C culture were transferred to 10 °C, there was a progressive increase in cold tolerance through three successive generations. Transfer of newly moulted pre-reproductive adults reared at 10 °C for three generations back to 20 °C resulted in a rapid loss of cold hardiness in their nymphal offspring.
3. In all generations reared at 10 °C, first born nymphs were more cold hardy than those born later in the birth sequence. The LT₅₀ of nymphs produced on the first day of reproduction in the first, second and third generations maintained at 10 °C were −14.8, −17.0 and −16.6 °C, respectively. Thereafter, nymphal cold hardiness decreased over the subsequent 14 days of reproduction in each generation at 10 °C with mean LT₅₀ values of −10.3, −12.6 and −14.8 °C, respectively. By contrast, the cold tolerance of first born nymphs of aphids reared continuously at 20 °C did not differ in comparison with later born siblings. The LT₅₀ of adult aphids was also unaffected by ageing.
4. The ecological relevance of these findings is discussed in relation to the overwintering survival of aphids such as S. avenae .     

Ibotenic Acid Analogues as Inhibitors of [3H]Glutamic Acid Binding to Cerebellar Membranes

Abstract: The L-[³H]glutamic acid binding capability of rat cerebellar membranes prepared with or without preincubation at 37°C followed by washing was investigated. The two preparations ( K_D = 820 nM, B_max= 54.5 pmol/mg protein; K_D = 509 nM, B_max=13.0 pmol/mg protein) showed no difference in specificity of the binding of the ibotenic acid analogues, consistent with the removal of an endogenous inhibitor by the preincubation at 37°C followed by washing. The order of potency of the ibotenic acid analogues as inhibitors of L-[³H]glutamic acid binding is different from the order of potency in vivo , suggesting that the binding sites found are different from the physiological glutamic acid receptor.     

Glucose fermentation to acetate and alanine in resting cell suspensions of Pyrococcus furiosus: Proposal of a novel glycolytic pathway based on 13C labelling data and enzyme activities

Abstract Suspensions of maltose-grown cells of the hyperthermophilic archaeon Pyrococcus furiosus , when incubated at 90°C with 35 mM [1-¹³C]glucose or [3-¹³C]glucose, consumed glucose at a rate of about 10 nmol min⁻¹ (mg protein)⁻¹. Acetate (10 mM), alanine (3 mM), CO₂ and H₂ were the fermentation products. The ¹³C-labelling pattern in alamine and acetate were analyzed. With [1-¹³C]glucose the methyl group of both alanine and acetate was labelled; with [3-¹³C]glucose only the carboxyl group of alanine was labelled whereas acetate was unlabelled. Extracts of maltose-grown cells contained glucose isomerase (12.8 U mg⁻¹, 100°C), ketohexokinase (0.23 U mg⁻¹, 100°C), and fructose 1-phosphate aldolase (0.06 U mg^{−1, 100°C). Enzymes catalyzing the formation of fructose 1,6-bisphosphate from fructose 1-phosphate or fructose 6-phosphate could not be detected. As publihed previously by our group and other authors P. furiosus also contains enzymes of glyceraldehyde conversion to 2-phosphoglycerate according to a non-phosphorylated Entner-Doudoroff pathway, of dihydroxyacetone phosphate conversion to 2-phosphoglycerate according to the Embden-Meyerhof pathway, and of 2-phosphoglycerate conversion - via pyruvate - to acetate and alanine. Based on the enzyme activities in P. furiosus , the following pathway for glucose degradation to alanine and acetate in cell suspensions is proposed which can explain the [13}C]glucose labelling data: glucose→ fructose → fructose 1- phosphate → dihydroxyacetone phosphate + glyceraldehyde and further conversion of both trioses to alanine and acetate via pyruvate.     

Macromolecular synthesis by yeasts under frozen conditions

Although viable fungi have been recovered from a wide variety of icy environments, their metabolic capabilities under frozen conditions are still largely unknown. We investigated basidiomycetous yeasts isolated from an Antarctic ice core and showed that after freezing at a relatively slow rate (0.8°C min⁻¹), the cells are excluded into veins of liquid at the triple junctions of ice crystals. These strains were capable of reproductive growth at −5°C under liquid conditions. Under frozen conditions, metabolic activity was assessed by measuring rates of [³H]leucine incorporation into the acid-insoluble macromolecular fraction, which decreased exponentially at temperatures between 15°C and −15°C and was inhibited by the protein synthesis inhibitor cycloheximide. Experiments at −5°C under frozen and liquid conditions revealed 2–3 orders of magnitude lower rates of endogenous metabolism in ice, likely due to the high salinity in the liquid fraction of the ice (equivalent of ≈ 1.4 mol l⁻¹ of NaCl at −5°C). The mesophile Saccharomyces cerevisae also incorporated [³H]leucine at −5°C and −15°C, indicating that this activity is not exclusive to cold-adapted microorganisms. The ability of yeast cells to incorporate amino acid substrates into macromolecules and remain metabolically active under these conditions has implications for understanding the survival of Eukarya in icy environments.     

Key Residues Defining the μ-Opioid Receptor Binding Pocket: A Site-Directed Mutagenesis Study

Abstract: Structural elements of the rat μ-opioid receptor important in ligand receptor binding and selectivity were examined using a site-directed mutagenesis approach. Five single amino acid mutations were made, three that altered conserved residues in the μ, δ, and κ receptors (Asn¹⁵⁰ to Ala, His²⁹⁷ to Ala, and Tyr³²⁶ to Phe) and two designed to test for μ/δ selectivity (Ile¹⁹⁸ to Val and Val²⁰² to Ile). Mutation of His²⁹⁷ in transmembrane domain 6 (TM6) resulted in no detectable binding with [³H]DAMGO (³H-labeled d -Ala², N -Me-Phe⁴,Gly-ol⁵-enkephalin), [³H]bremazocine, or [³H]ethylketocyclazocine. Mutation of Asn¹⁵⁰ in TM3 produces a three- to 20-fold increase in affinity for the opioid agonists morphine, DAMGO, fentanyl, β-endorphin_1–31, JOM-13, deltorphin II, dynorphin_1–13, and U50,488, with no change in the binding of antagonists such as naloxone, naltrexone, naltrindole, and nor-binaltorphamine. In contrast, the Tyr³²⁶ mutation in TM7 resulted in a decreased affinity for a wide spectrum of μ, δ, and κ agonists and antagonists. Altering Val²⁰² to Ile in TM4 produced no change on ligand affinity, but Ile¹⁹⁸ to Val resulted in a four- to fivefold decreased affinity for the μ agonists morphine and DAMGO, with no change in the binding affinities of κ and δ ligands.     

Pharmacology and Regional Distribution of the Binding of 6-[3H]Nitro-7-Sulphamoylbenzo[f]-Quinoxaline-2,3-Dione to Rat Brain

Abstract: 6-Nitro-7-sulphamoylbenzo[ f ]quinoxaline-2,3-dione (NBQX) is a competitive antagonist selective for α-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors. Here we report the pharmacological characteristics and anatomical distribution of [³H]NBQX binding to rat brain. The association rate of [³H]NBQX to rat cerebrocortical membranes was rapid, with peak binding occurring within 10 min at 0°C. The off-rate was also rapid, with near-complete dissociation of the radioligand within 5 min of addition of 1 m M unlabelled l -glutamate. [³H]NBQX bound to a single class of sites with K _D and B _max values of 47 n M and 2.6 pmol mg⁻¹ of protein, respectively. The rank order of inhibition of [³H]NBQX binding by AMPA receptor ligands was NBQX ≫ 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) ≥ ( S )-5-fluorowillardiine ≥ AMPA ≫ l -glutamate. The chaotrope KSCN had no effect on the IC₅₀ value of unlabelled NBQX displacement of [³H]NBQX binding. The kainate receptor-selective ligands NS102 and kainate were only very weak displacers. It is interesting that NBQX and CNQX displaced significantly more [³H]NBQX than any of the agonists tested. Autoradiographic analysis of the binding of [³H]NBQX to coronal sections showed a distribution compatible with that of [³H]AMPA binding. These data indicate that [³H]NBQX provides a useful novel tool to characterise the antagonist binding properties of AMPA receptors.     

Measurement of the Membrane Potential of Isolated Nerve Terminals by the Lipophilic Cation [3H]Triphenylmethylphosphonium Bromide

Abstract: The lipophilic cation [³H]triphenylrnethylphosphonium bromide ([³H]TPMP⁺) was investigated as a measure of the membrane potential of synaptosomes. Conditions under which [³H]TPMP⁺ achieved an equilibrium distribution were tested. The toxicity of TPMP has been studied relative to its inhibitory effects on [³H]y-aminobutyric acid ([³H]GABA) transport. In some experiments the distribution of ⁸⁶RbZ⁺ and [³H]TPMP⁺ was changed upon incubation in the presence of elevated levels of K⁺, ouabain, or KCN, or at 0°C in a way that would be expected from the membrane potential. In normal incubation conditions a membrane potential of ∼−60 mv was calculated.