Effect of rearing density on thyroid and interrenal gland activity and plasma and hepatic metabolite levels in rainbow trout, Salmo gairdneri Richardson

There were significant inverse correlations between rearing density of rainbow trout, Salmo gairdneri Richardson, and final body weight, plasma L-thyroxine (T₄), trüodo-L-tryronine (T₃), cortisol and protein concentrations, plasma T₄/T₃ ratios and thyroid epithelial cell height. In addition, hepatosomatic indices and plasma free fatty acid concentrations were higher in fish reared at low (134 g 1⁻¹) density compared with groups reared at medium (210g1⁻¹) or high density (299g 1⁻¹), and the post-feeding (3.5-4h) elevation in plasma glucose and triglyceride levels evident in trout maintained at low rearing density was not found in those fish reared at higher densities. There were no significant effects of rearing density on hematocrit, carcass composition, hepatic glycogen and lipid levels and interregnal nucleus size.     

Changes in Glycoprotein Metabolism in the Cerebral Cortex Following First Exposure of Dark-reared Rats to Light

Abstract: The level of incorporation of [³H]fucose or [³H]lysine into subcel-lular fractions of the visual and motor cortices of 50-day-old dark-reared (D) and light-exposed (L) rats was determined. No differences were found between D and L rats in the incorporation of either precursor into subcellular fractions of the motor cortex, or in any fraction of the visual cortex except the synaptic-membrane fraction. After a 3-h light exposure the incorporation of [³H]fucose into the visual cortex synaptic-membrane fraction was elevated (L/D = 136%). Incorporation of [³H]lysine was elevated in the visual cortex synaptic-membrane fraction of L compared to D rats after a 1-h exposure (L/D = 118%). However, after a 3-h exposure the incorporation was depressed in this fraction (L/D = 79%). No differences could be found in the levels of activity of fucosyl transferase following first exposure to light but dark-rearing itself resulted in increased enzyme activity in the motor cortex compared to normal controls. First exposure of 20-day-old dark-reared rats to light led to an increase in the incorporation of [³H]fucose into soluble glycoproteins of both the visual and motor cortex and into particulate glycoproteins of the visual cortex only. These results are in contrast with those found with 50-day-old animals and suggest that the effects of light-exposure on [³H]fucose incorporation may be age-dependent.     

INCREASED BINDING OF [3H]COLCHICINE TO VISUAL CORTEX PROTEINS OF DARK-REARED RATS ON FIRST EXPOSURE TO LIGHT

Abstract— The binding of [³H]colchicine (or a functionally similar metabolite) to acid-insoluble material in vivo was measured in the motor and visual cortices of littermates rats which were either dark-reared (D), exposed to light for 3 h or 24 h (L), or raised normally (N) in 12 h light/12 h dark animal house conditions. No significant differences were found in the binding in the motor cortex of the 3 h or 24 h L, D or N animals, but in the visual cortex after 3 h of light exposure a 23% elevation in binding was measured in L compared with D animals and a small though non-significant (10%) increase in binding was also observed in this region in L compared with N animals. After 24 h of light exposure, binding of the label in the L animals fell near to that of the N and D animals. The results of vinblastine precipitation experiments suggested that much of the radioactivity was bound to the protein tubulin, and this was confirmed when no increased binding of an analogue of colchicine, lumi-colchicine, was observed after 3 h of light exposure in L compared with D animals. It is suggested that these experiments show that colchicine can be used as a marker for changes in the tubulin population in light exposed animals, and demonstrate the transient nature of the increase in tubulin quantity, as opposed to a lasting effect on its synthesis. Further, they argue strongly in support of the idea that a component of protein flow from neuronal cell body to axons and dendrites in light exposed animals, is subject to environmental modification.     

Simultaneous Measurement of Nitrite and Nitrate Levels as Indices of Nitric Oxide Release in the Cerebellum of Conscious Rats

Abstract: We examined the modulation of nitric oxide production in vivo by measuring levels of nitrite (NO₂⁻) and nitrate (NO₃⁻) in the dialysate of the cerebellum in conscious rats, by using an in vivo brain microdialysis technique. The levels of both NO₂⁻ and NO₃⁻ were decreased by the intraperitoneal injection of N ^G-nitro- l -arginine methyl ester, an inhibitor of nitric oxide synthase, whereas N ^G-nitro- d -arginine methyl ester had no effect. l -Arginine by itself increased NO₂⁻ and NO₃⁻ levels and diminished the reduction of their levels caused by N ^G-nitro- l -arginine methyl ester. Direct infusion of l -glutamate, N -methyl- d -aspartate, or KCl into the cerebellum through a dialysis probe resulted in an increase in NO₂⁻ and/or NO₃⁻ levels. The effects of N -methyl- d -aspartate and KCl were dependent on extracellular calcium. Furthermore, the stimulatory effects of l -glutamate and N -methyl- d -aspartate were inhibited by N ^G-nitro- l -arginine methyl ester and (±)-3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP), an N -methyl- d -aspartate receptor antagonist. These results suggest that NO₂⁻ and NO₃⁻ levels may be related to nitric oxide production in vivo.     

Effect of chlorination on β-D-galactosidase activity of sewage bacteria and Escherichia coli

The effect of chlorine on β- D- galactosidase activity of sewage bacteria and Escherichia coli was studied. β- D- galactosidase activity of sewage was more resistant to chlorine than faecal coliform cultivability. At low initial dosage (0·05 mg Cl₂ l⁻¹) neither cultivability (colony-forming units (cfu)), nor enzyme activity of E. coli suspensions were severely impaired. When initial chlorine concentration was increased to 0·1 mg Cl₂ l⁻¹, the cfu number decreased whereas enzyme activity remained high, i.e. the enzyme activity calculated cfu⁻¹ increased. At higher chlorine doses both cfu and enzyme activity were reduced, but non-cultivable cells retained assayable activity after chlorination. Mean values of the enzyme activity calculated cfu⁻¹ decreased when the chlorine dosage was increased from 0·1 to 0·5 mg Cl₂ l⁻¹, but were not significantly different ( P > 0·05) for dosages of 0·2–0·7 mg Cl₂ l⁻¹. After chlorination, β- D- galactosidase activity of E. coli was less reduced than cfu and direct viable count numbers, but more reduced than 5-cyano-2-3, ditolyl tetrazolium chloride and total cell counts, and the enzyme activity represented an alternative activity parameter of chlorinated samples.     

Quantitative Ultrastructural Investigations on Synaptogenesis in the Cerebellum and the Optic Tectum of Light-Reared and Dark-Reared Rainbow Trout (Salmo gairdneri Rich.)

The morphogenetic differentiation of synapses in the cerebellum and the optic tectum of darkand light- reared rainbow trout was investigated at critical stages of development. During normal differentiation the cerebellum is characterized by the appearance of 'indented', spinelike synapses. This type of synapses increases with age and prevails from day 60 on. At the same time the number of 'flat' synapses decreases. In the cerebellum the highest synaptic density (123 ± 12 synapses/1,000 μm²) is reached 30 days after hatching when the larvae begin to swim. The optic tectum is characterized by a preponderance of flat synapses in early postnatal and adult life; maximal synaptic density (66 ± 5 synapses/1,000 μm²) is reached 60 days after hatching when the larvae have reached optimal visual acuity.
Light deprivation causes a considerable and significant reduction in the number of synapses per unit area in the cerebellum and the optic tectum. The length of synaptic contacts do not change. If light-deprived, the density of synaptic vesicles decreases significantly in the optic tectum of a 25-day-old trout (74 ± 3 instead of 132 ± 7 vesicles/μm²). In the cerebellum this effect is absent.     

Comparison of Voltage-Dependent 45Ca2+ Uptake Rates by Synaptosomes Isolated from Rat Brain Regions

Abstract: ⁴⁵Ca²⁺ uptake by synaptosomes isolated from cerebral cortex, cerebellum, midbrain, and brain stem of male Sprague-Dawley rats was measured at 1-, 3-, 5-, 15-, 30-, and 60-s time periods. The fastest rate of depolarization-dependent calcium uptake occurred in each brain region between 0 and 1 s. Uptake rates dropped off quickly with 3–5-s rates at approximately 15–20% of those observed at 0–1 s in cerebral cortex, cerebellum, and midbrain. Uptake rates at the 1–3-s interval were maintained at a relatively high rate in these three brain regions suggesting mixed fast- and slow-phase processes. The magnitude and rate of ⁴⁵Ca²⁺ uptake were similar in synaptosomes from cerebral cortex, cerebellum, and midbrain but were significantly less in brain stem synaptosomes. These results suggest a fast and a slow component to voltage-dependent ⁴⁵Ca²⁺ uptake by presynaptic nerve terminals from various brain regions.     

Differential Interaction of 1-Methyl-4-Phenylpyridinium Ion with the Putatively Vesicular Binding Site of [3H]Tyramine in Dopaminergic and Nondopaminergic Brain Regions

Abstract: The neurotoxin 1-methyl-4-phenylpyridinium ion (MPP⁺) in the brain striatum has recently been shown to bind at a putatively vesicular site labeled by [³H]tyramine ([³H]TY). Whereas in the rat and mouse striatum MPP⁺ antagonized TY binding competitively, in the cerebellum there was a mixed-type antagonism, which suggests the simultaneous occupancy of two different sites. K _i values from displacement curves revealed a fourfold difference in the affinity of MPP⁺ for TY sites in the two brain regions. The degeneration of central noradrenergic terminals induced by an intraperitoneal injection of the toxin N -(2-chloroethyl)- N -ethyl-2-bromobenzylamine in rats decreased by 80% the maximal number of cerebellar TY binding sites, while not affecting striatal binding. Furthermore, guanethidine, a marker for noradrenaline (NA) vesicles, potently inhibited TY binding in NA-innervated regions, such as the cerebellum and the parietal cortex, and poorly in the striatum. It is concluded (a) that both MPP⁺ and TY may also label NA vesicles and (b) that the vesicular carriers for dopamine and NA have different characteristics, which may underlie a regional specificity in the rate of endovesicular sequestration of MPP⁺, with either neurodegenerative or neuroprotective consequences, depending on the brain area involved.     

Sodium-dependent dynamic assembly of membrane complexes in sodium-driven flagellar motors

The bacterial flagellar motor is driven by the electrochemical potential of specific ions, H⁺ or Na⁺. The motor consists of a rotor and stator, and their interaction generates rotation. The stator, which is composed of PomA and PomB in the Na⁺ motor of Vibrio alginolyticus , is thought to be a torque generator converting the energy of ion flux into mechanical power. We found that specific mutations in PomB, including D24N, F33C and S248F, which caused motility defects, affected the assembly of stator complexes into the polar flagellar motor using green fluorescent protein-fused stator proteins. D24 of PomB is the predicted Na⁺-binding site. Furthermore, we demonstrated that the coupling ion, Na⁺, is required for stator assembly and that phenamil (an inhibitor of the Na⁺-driven motor) inhibited the assembly. Carbonyl cyanide m -chlorophenylhydrazone, which is a proton ionophore that collapses the sodium motive force in this organism at neutral pH, also inhibited the assembly. Thus we conclude that the process of Na⁺ influx through the channel, including Na⁺ binding, is essential for the assembly of the stator complex to the flagellar motor as well as for torque generation.     

Kinetics, Pharmacology, and Autoradiographic Distribution of l-[3H]Nitroarginine Binding Sites in Rat Cerebellum

Abstract: The kinetics and pharmacology of N ^G-nitro- l -[2,3,4,5-³H]arginine ( l -[³H]NOARG) binding to rat cerebellum were investigated using in vitro radioligand binding. Specific l -[³H]NOARG binding in cerebellum was of nanomolar affinity, reversible, saturable, and best fit to a single-site model. Specific binding was Ca²⁺ dependent and sensitive to pH (with an optimum of 5.5–7.0). Added calmodulin (1.5–40 µg/ml) had no influence on specific l -[³H]NOARG binding. However, the calmodulin antagonists W-5, W-13, and calmidazolium inhibited l -[³H]NOARG binding with IC₅₀ values in the micromolar range, and calmodulin (10 µg/ml) competitively reversed this inhibition. Nitric oxide synthase (NOS) inhibitors ( N ^G-nitro- l -arginine methyl ester and N ^G-monomethyl- l -arginine acetate) and l -arginine displaced l -[³H]NOARG binding with IC₅₀ values in the nanomolar range, whereas d -arginine and basic amino acids ( l -lysine and l -histidine) displaced l -[³H]NOARG binding with IC₅₀ values in the millimolar range. A comparison of the NOS functional assay with l -[³H]NOARG binding in rat cerebellum showed similar profiles of Ca²⁺ dependency and inhibitory kinetics. Quantitative autoradiographic distribution of l -[³H]NOARG binding sites was significantly higher in the molecular layer than in the granular layer of cerebellum. These studies confirm the potential use of l -[³H]NOARG binding to study the regional distribution and functional properties of NOS.     

Regional Calcium Accumulation and Cation Shifts in Rat Brain by Kainate

Abstract: Following local application of kainic acid, changes in the contents of Na⁺, K⁺, Ca²⁺, and Mg²⁺ of the striatum, cerebellum, and hippocampus of the rat were observed at various times after surgery. Within 1 h the levels of K⁺ decreased 20% whereas the levels of Na⁺ and Ca²⁺ increased at least 50% and 20%, respectively. These changes persisted for more than 8 weeks. Ca²⁺ levels rose further, to more than 10-fold during 8 weeks. The Mg²⁺ levels were slightly and only transiently decreased. Unilateral injections of kainate into the striatum affected the contents of these cations not only in this area, but also in the overlying cerebral cortex, the olfactory tubercle, and the ipsilateral substantia nigra. The Ca²⁺ increases were less when rats were kept on a diet deficient in Ca²⁺ and vitamin D. ⁴⁵Ca²⁺, intravenously administered, accumulated significantly more in the kainate-lesioned striatum and substantia nigra than in the homotopic contralateral areas. Electron microscopic examination of the localization of Ca²⁺ with the oxalate-pyroantimonate technique showed the appearance of extracellularly located deposits and the accumulation of Ca²⁺ in (possibly degenerating) myelinated axons in kainate-lesioned striata. This study provides evidence that calcification of cerebral tissue is closely associated with neurodegenerative processes and shows that kainate may serve as a tool to elucidate the mechanism of brain calcification. The results are discussed in relation to idiopathic calcinosis (striopallidodentate calcinosis, Fahr's disease).     

Denitrification in stream epilithon: Seasonal variation in Gelbæk and Rabis Bæk, Denmark

Abstract Seasonal variations of denitrification activity were determined and compared with various environmental parameters in undisrupted epilithon communities (biofilm) from two Danish lowland streams. In the nutrient-rich Gelbæk, denitrification activity followed extensive changes in biofilm thickness, dry weight and chlorophyll a content during the season. The absolute maximum of denitrification (1.4 mmol N m⁻² d⁻¹, dark incubation) was recorded when the biofilm was best developed in the spring (April and May). Activity decreased dramatically after most of the biofilm suddenly disintegrated and peeled off in early summer. Photosynthetic O₂ production was an important controlling factor of denitrification on a diurnal scale, since the activities were always 2–3 fold lower in the light (50 μE m⁻² s⁻¹) than in the dark. In the more nutrient-poor Rabis Bæk, the biofilm was much less developed and denitrification activities were relatively small (maximum of 0.2 mmol N m⁻² d⁻¹ under dark incubation). The results indicate that productivity of the micro-algae regulates both the seasonal and diurnal patterns of denitrification in the biofilms.     

Alterations in beta 1- and beta 2-adrenergic receptor density in the cerebellum of aging rats

Abstract: The densities of β¹, and β₂-adrenergic receptors were determined in homogenates of cerebral cortex and cerebellum of rats between 3 and 14 mo of age. No change in either receptor population occurred in the cortex during this period. In the cerebellum, a 20–25% decrease in the density of β₂, receptors and a 3509% increase in the density of β₁, receptors occurred. The increase in β₁ receptors in the cerebellum may be the result of a decrease in the function of the noradrenergic projections from the locus coeruleus which synapse on cerebellar Purkinje cells.     

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.     

Involvement of Different Types of Voltage-Sensitive Calcium Channels in the Presynaptic Regulation of Noradrenaline Release in Rat Brain Cortex and Hippocampus

Abstract: Transmitter release at the nerve terminal is mediated by the influx of Ca²⁺ through voltage-sensitive calcium channels (VSCCs). Many types of VSCCs have been found in neurons (T, N, L, and P), but uncertainty remains about which ones are involved in neuronal excitation-secretion coupling. Specific ligands for the L- and N-type VSCCs were used to determine which of these subtypes might be involved in the K⁺-evoked [³H]noradrenaline release from superfused rat brain cortical and hippocampal synaptosomes. In cortical presynaptic terminals the 1,4-dihydropyridine agonist Bay K 8644 enhanced the K⁺ (15 m M )-evoked [³H]noradrenaline release. This effect was reversed by the 1,4-dihydropyridine antagonists nimodipine and nitrendipine. The L-type VSCC ligands had no effect on hippocampal synaptosomes. In contrast, the N-type VSCC blocker ω-conotoxin markedly reduced the K⁺-evoked [³H]noradrenaline release in nerve terminals from both regions. Inhibition was greater in hippocampal synaptosomes. When applied together the inhibitory actions of nimodipine and ω-conotoxin were approximately additive. These findings indicate that both L- and N-type VSCCs participate in noradrenaline release in rat brain cortex and suggest that noradrenergic terminals in the two regions examined may have distinct populations of VSCCs: L type in cortex and N type in hippocampus.     

The role of sulphur compounds for breeding success of Ips typographus L. (Col., Scolytidae) on Norway Spruce (Picea abies [L.] Karst.)

Bark beetles, especially Ips typographus L. represent a severe biotic threat for spruce ( Picea abies [L.] Karst.) at low altitudes in Europe. We compared sulphur (total S, SO₄²⁻, glutathione, cysteine, methionine), nitrogen (total N, NO₃⁻, total protein, free amino acids), carbon, total phosphorus and PO₄³⁻, tree vigour index (TVI) and water content of the phloem after felling, and their dependent changes (tdc) with the breeding success of I. typographus . Twenty trees were classified according to age (34/90 years) and crown density (high/intermediate/low). Water content was higher in young trees than in old trees, higher in the crown than at breast height, and decreased significantly within the 8-week study period. In old trees, breeding success, length of mother galleries and SO₄²⁻ were significantly higher, while total protein, NO₃⁻ and water content were significantly lower than in young trees. Trees with intermediate crown density provided the best breeding success for I. typographus and had significantly higher arginine content and C/N ratio as well as low amounts of phosphate and glutamine. During the period of bark beetle breeding, total sulphur, glutathione, protein, NO₃⁻, aspartate, glutamine, glutamate, arginine and γ-aminobutyrate decreased significantly. The results support previous investigations that I. typographus develops best in physiologically weakened trees.     

Interaction of potassium ions and gibberellin in the control of hypocotyl growth in Amaranthus caudatus

Potassium promotes growth in several plant tissues. Elongation growth of the hypocotyls of Amaranthus caudatus L. ev. Lalsag is mainly controlled by gibberellins, but K⁺ also promotes growth. In the present study the interaction of K⁺ with gibberellin (GA₃) and chlorocholine chloride (CCC) has been investigated. When K⁺ was applied externally in the dark, hypocotyl growth was promoted in the seedlings. External application of GA₃ did not promote growth in the dark. GA₃ was effective in the light and K⁺ was synergistic with GA₃ in promoting elongation. Application of CCC in the dark makes the seedlings sensitive to GA₃. The inhibition of growth by CCC was also reversed by K⁺. The results indicate a possible role of K⁺ in GA₃ induced elongation of hypocotyls.     

Colonization of Vibrio pelagius and Aeromonas caviae in early developing turbot (Scophthalmus maximus L.) larvae

Polyclonal antisera made in rabbits against whole washed cells of Vibrio pelagius and Aeromonas caviae were used for detection of these bacterial species in the rearing water and gastrointestinal tract of healthy turbot ( Scophthalmus maximus ) larvae exposed to V. pelagius and/or Aer. caviae . The results demonstrated that this method is suitable for detection of V. pelagius and Aer. caviae in water samples and larvae at population levels higher than 10³ ml⁻¹ and 10³ larva⁻¹. Populations of aerobic heterotrophic bacteria present in the gastrointestinal tract of turbot larvae, estimated using the dilution plate technique, increased from approximately 4 × 10² bacteria larva⁻¹ on day 3 post-hatching to approximately 10⁵ bacteria fish⁻¹ 16 days post-hatching. Sixteen days after hatching, Vibrio spp. accounted for approximately 3 × 10⁴ cfu larva⁻¹ exposed to V. pelagius on days 2, 5 and 8 post-hatching. However, only 10³ of the Vibrio spp. belonged to V. pelagius . When larvae were exposed to Aer. caviae on day 2 post-hatching, the gut microbiota of 5-day old larvae was mainly colonized by Aeromonas spp. (10⁴ larva⁻¹), of which 9 × 10³ belonged to Aer. caviae . Later in the experiment, at the time when high mortality occurred, 9 × 10⁵ Aer. caviae were detected. Introduction of V. pelagius to the rearing water seemed to improve larval survival compared with fish exposed to Aer. caviae and with the control group. It was therefore concluded that it is beneficial with regard to larval survival to introduce bacteria ( V. pelagius ) to the rearing water.     

Influence of Melatonin on the Rate of Rana pipiens Tadpole Metamorphosis In Vivo and Regression of Thyroxine-Treated Tail Tips In Vitro

Metamorphosis of Rana pipiens tadpoles may be retarded when the light phase of the light/dark (LD) cycle is shortened or when thyroxine (T₄) is given in the dark because melatonin peaks during the dark. Injection of premetamorphic tadpoles in spontaneous metamorphosis with melatonin (15 μg) retarded tail growth and hindlimb development on 18L:6D but had no significant effect on 6L:18D. During induced metamorphosis (30 μg/liter T₄), melatonin injections retarded tail resorption on 18L:6D and accelerated it on 6L:18D, but did not affect the hindlimb. When melatonin was injected during T₄ immersion at different times in the photophase on 18L:6D (L onset 0800 hr), tail regression was retarded by melatonin at 1430 or 2030 hr. At 0830 hr, shrinkage of tail length was accelerated whereas tail height was not affected. Tail tips in vitro induced to resorb by 0.2 μg/ml T₄ in Niu-Twitty solution regressed more slowly in the presence of melatonin (10 or 15 μg/ml) than with T₄ alone on both 6L:18D and 18L:6D. The findings implicate melatonin in LD cycle effects on tadpole metamorphic rate in vivo , show the importance of the time of melatonin injections, and indicate that melatonin antagonizes the metamorphic action of T₄ at the tissue level.     

Aged endothelial nitric oxide synthase knockout mice exhibit higher mortality concomitant with impaired open-field habituation and alterations in forebrain neurotransmitter levels

Endothelial nitric oxide synthase (eNOS) has been implicated in various brain and peripheral pathologies such as renal failure, heart failure or stroke. Consequently, the mortality rate of aged eNOS knockout mice (eNOS^–/–) was higher than that of age-matched (18–22 months old) controls. Only seven of the original 14 eNOS^–/– animals that participated in the study reached the age of 18 months or older, whereas no control mice died during this life span. In order to assess the behavioral and neurochemical consequences of chronic eNOS deficiency we examined whether the surviving aged eNOS^–/– mice showed changes in terms of motor, emotional, exploratory and neurochemical parameters. Aged eNOS^–/– mice showed reduced exploratory activity in the open-field with no habituation observable neither within sessions nor after repeated exposures. Pole test performance of eNOS^–/– mice was comparable to controls. In the elevated plus-maze eNOS^–/– mice did not differ from controls in terms of time spent in and entries into arms, but showed less locomotion on the open arms. The most prominent neurochemical alterations in the forebrains of aged eNOS^–/– mice were: (a) increased acetylcholine levels in the neostriatum; (b) decreased noradrenaline concentrations in the ventral striatum; and (c) lower serotonin levels in the frontal cortex and ventral striatum. The present findings suggest that mice which survived chronic eNOS-deficiency into old age, show some behavioral and neurochemical phenotypes distinct from adult eNOS^–/– mice.