Localization of NADPH-diaphorase in the goldfish brain

The distribution of NADPH-diaphorase positive neurons was studied by using the enzyme histochemical method. Numerous neurons were labeled in different brain areas of the goldfish and their distribution showed some differences in comparison with other studied teleosts, indicating a species-specific pattern of NADPH-diaphorase distribution as observed in mammals. The localization of NADPH-diaphorase in the thalamic nuclei, in the paraventricular organ, in the inferior hypothalamic lobe, in the periventricular neurons of the optic tectum, in the nucleus isthmi and in the mesencephalic reticular formation was comparable to the one observed in other teleosts. In addition in the goldfish the telencephalic neurons of the pars centralis and lateralis of the area dorsalis, the habenular neurons, the bipolar neurons of the central grey layer of the optic tectum and the motor neurons of the hypertrophied vagal lobe were labeled. The localization of NADPH-diaphorase positive neurons, compared to the distribution of cholinergic neurons described in fish, indicated that the production of nitric oxide was prevailing in the brain areas where cholinergic circuits are active.     

Circadian oscillations of DNA synthesis in rat brain

The possibility that the synthesis of brain DNA undergoes a circadian fluctuation was examined in male adult Wistar rats, kept under natural lighting conditions or born and raised under artificial lighting conditions. Groups of rats were taken every 4 h during the 24 h, injected subcutaneously with [methyl-3H]thymidine and killed 4 h later. By cosinor analysis, the DNA specific activity of cerebral hemispheres and brainstem was found to show a significant 24 h rhythm with the peak at the beginning of the dark period (waking period). By contrast, in kidney, the peak of the circadian rhythm of DNA specific activity occurred during the light period (sleep period), in agreement with literature data. On the other hand, in 4-week-old rats, born and raised in artificial lighting conditions, brain DNA specific activity followed a 12 h rhythm, in agreement with the lack of a significant diurnal oscillation of the sleep--waking structure. It is concluded that brain DNA synthesis undergoes a circadian fluctuation in association with the circadian rhythm of waking.     

Ultrastructural localization of NADPH-diaphorase in the goldfish brain

The ultrastructural localization of NADPH-diaphorase was studied in the goldfish brain by means of the tetrazolium salt technique. The reaction product was found to be bound to the endoplasmic membranes of neurons in different brain areas. In the synaptic structures both pre and post-synaptic profiles showed the reaction product. Furthermore non-neuronal structures were intensely labeled. Endothelial cells revealed the membranous localization of NADPH-diaphorase and the glial cells of the hypothalamic nuclei and of the paraventricular organ were labeled. In some cases the reaction product was seen also in oligodendrocytes and in microglial cells.     

Characteristics, day-night changes, subcellular distribution and localization of melatonin binding sites in the goldfish brain

Melatonin binding sites in the goldfish brain were characterized by radioreceptor assay using 2-[¹²⁵I]iodomelatonin as the radioligand. Specific binding of 2-[¹²⁵I]iodomelatonin was rapid, stble, saturable and reversible. Saturation experiments demonstrated that 2-[¹²⁵I]iodomelatonin binds to a single class of receptor site with an affinity constant (K_rmd) of 29.8±0.7pM and a total binding capacity (B_max)of11.47 ± 0.33fmol/mg protein at mid-dark, the B_max value decreased significantly to 7.90±0.23fmol/mg protein (Pˇ0.01) with no significant variation in the K_d value (33.8±1.5pM). Competition experiments revealed the following order of pharmacological affinities: 2-iodomelatonin>melatonin> 6-hydroxymelatonin> N-acetyl-5-hydrxytryptamine> 5-methoxytryptamine> 5-methoxytryptophol> 5-methoxyindole-3-acetic acid. 5-Hydroxytryptamine, 5-hydroxytryptophol, 5-hydroxyindole-3-acetic acid, norepinephrine and acetylcholine exhibited no inhibition. Subcellular distribution of melatonin binding sites was demonstrated to be greatest in the P₂ and P₃ fractions as compared with the P₁ fraction. Localization of melatonin binding sites in discrete brain areas was determined to be highest in the optic tectum-thalamus and hypothalamus, intermediate in the telencephalon, cerebellum and medulla oblongata, and lowest in the olfactory bulbs and pituitary gland. These results suggest that characteristics of melatonin receptors are highly conserved during evolution and that in this species melatonin plays neuromodulatory roles in the central nervous system through specific receptors.     

Phylogeny of histamine in vertebrate brain

Measurement of histamine and its metabolizing enzymes in a variety of chordate species indicated that histamine and histamine methyltransferase were present in brain of all vertebrate species with a recognizable brain structure. Diamine oxidase was absent in mammalian brain but was present in brain of lower vertebrates. Histamine levels were especially high in amphibia and fish brains, in which the phylogenetically newer parts of the brain were less well-developed. In the spiny dogfish (as in mammals), brain histamine levels were highest in the midbrain regions. In contrast to brain, histamine levels were low in muscle, skin and intestine of all aquatic species.     

Reversible changes in goldfish brain polyamine concentrations and synthetic enzymes after cold exposure

Exposure of goldfish to the cold (5 degrees C) caused a sharp increase in brain putrescine level during the first week. Such increase continued at a minor rate for the whole period of exposure (2 months). In contrast, the content of spermidine and spermine remained unchanged. Putrescine increase was concomitant with a remarkable rise in ornithine decarboxylase activity (ODC), which reached a maximum stimulation after 1 week of cold exposure, and declined thereafter, remaining significantly higher than the control for the entire period of study. Cold exposure caused also a reduction of S-adenosylmethionine decarboxylase (AdoMetDC) activity and an increase of ornithine level, whereas methionine content was unchanged. When fish exposed to cold temperature were returned to 20 degrees C, the modifications observed on brain polyamine metabolism were completely reversed. Supported by previous observations, our results suggest that the changes in the polyamine metabolism induced in goldfish brain by cold exposure could represent an homeostatic mechanism carried out by the goldfish to minimize the possible effects of thermal changes.     

Melatonin signal transduction in the goldfish, Carassius auratus

Generation and reception of melatonin signals in the goldfish, Carassius auratus, are reviewed. The photoreceptive pineal gland of the goldfish generates circulating melatonin rhythms according to a given photoperiod under light-dark cycles and in a circadian manner under continuous dark conditions. Melatonin is also produced in the retina in a similar fashion. Melatonin produced in the pineal gland and retina is considered to act as internal zeitgeber in the brain and retina, respectively, controlling various physiological events via specific melatonin binding sites that are coupled with G protein. The goldfish exhibit clear diurnal locomotor activity rhythms under light-dark cycles and free-running rhythms under constant conditions. However, the relationship between melatonin and locomotor activity rhythms in the goldfish remains unclear. Further studies should be required to demonstrate the roles of melatonin in the circadian system in this species.     

Melatonin signal transduction in the goldfish, Carassius auratus

Generation and reception of melatonin signals in the goldfish, Carassius auratus, are reviewed. The photoreceptive pineal gland of the goldfish generates circulating melatonin rhythms according to a given photoperiod under light-dark cycles and in a circadian manner under continuous dark conditions. Melatonin is also produced in the retina in a similar fashion. Melatonin produced in the pineal gland and retina is considered to act as internal zeitgeber in the brain and retina, respectively, controlling various physiological events via specific melatonin binding sites that are coupled with G protein. The goldfish exhibit clear diurnal locomotor activity rhythms under light-dark cycles and free-running rhythms under constant conditions. However, the relationship between melatonin and locomotor activity rhythms in the goldfish remains unclear. Further studies should be required to demonstrate the roles of melatonin in the circadian system in this species.     

Circadian variation in heart-rate variability in localization-related epilepsy

PURPOSE: Case-control studies of sudden unexpected death in epilepsy (SUDEP) have reported that SUDEP is more likely to occur during sleep and thus presumably during night hours. The circadian variation of heart-rate variability (HRV) might be of relevance to this risk. We examined night versus daytime HRV in patients with newly diagnosed and refractory localization-related epilepsy, assessing the effects of drug treatment and epilepsy surgery on the night/daytime HRV ratio. METHODS: We used spectral analysis to assess HRV and calculated the night-time (00.00-05.00)/daytime (07.30-21.30) ratio of HRV in 14 patients with newly diagnosed localization-related epilepsy before and during carbamazepine (CBZ) treatment and in 21 patients with temporal lobe epilepsy before and after epilepsy surgery. Both groups were compared with age- and sex-matched controls. RESULTS: No significant differences were found from controls in the night/daytime ratios of HRV whether compared before or after initiation of treatment with CBZ in newly diagnosed epilepsy patients. When patients were used as their own controls, night/daytime ratios of standard deviation of RR intervals (p = 0.04) and total power (p = 0.04) were significantly lower during treatment than before. Compared with those of controls, the night/daytime ratios were lower in epilepsy surgery patients before surgery [low-frequency power (p = 0.04); high-frequency power (p = 0.04)]. Night/daytime ratios did not change significantly after surgery. Conclusions: The HRV of the patients was more affected during night-time when the risk of SUDEP seems to be highest in such patients.     

Circadian variation of nitric oxide synthase activity and cytosolic protein levels in rat brain

The circadian variation of nitric oxide synthase (NOS) activity and cytosolic protein content in the cerebellum, brainstem, hypothalamus, hippocampus, and the remainder of the brain were studied in rats. Both NOS activity and cytosolic protein concentrations were the highest during the dark period and lowest in the light period. Hypothalamic NOS activity exhibited the most pronounced change in activity with time increasing by 120% from mid-light to mid-dark.     

Coefficient of variation of R-R intervals in severe brain damage

The coefficient of variation of R-R intervals (CVRR) was studied in 18 children having severe brain damage with a mean ± standard deviation (S.D.) age of 8.4 ± 5.9 years, who were divided into ten patients complicated with respiratory insufficiency (RI group) and eight patients with severe athetotic cerebral palsy (SA group). CVRR was obtained in the resting supine position, and was compared with that in 22 neurologically normal controls. CVRR in the RI group (mean ± S.D., 2.19 ± 1.28%) was significantly lower than that in controls (5.56 ± 1.53%), while CVRR in the SA group (11.30 ± 3.91%) was significantly higher than that in controls (both P < 0.01, ANOVA). In particular, the four patients with brain death showed extremely low CVRR of 1.00–1.29%. Since CVRR was 4.09% in the patient aged 4 years with birth injury of the upper cervical spinal cord causing absence of spontaneous respiration, the extremely low CVRR in patients with brain death may be directly related to brainstem dysfunction. The cause of the high CVRR in the SA group was not determined. Thus, CVRR may be useful for quantitative evaluation of severe neurological disorder.     

Decreased histamine H1 receptor binding in the brain of depressed patients

The central histaminergic neuron system modulates the wakefulness, sleep-awake cycle, appetite control, learning and memory, and emotion. Previous studies have reported changes in neuronal histamine release and its metabolism under stress conditions in the mammalian brain. In this study, we examined, using positron emission tomography (PET) and [(11)C]-doxepin, whether the histaminergic neuron system is involved in human depression. Cerebral histamine H1 receptor (H(1)R) binding was measured in 10 patients with major depression and in 10 normal age-matched subjects using PET and [(11)C]-doxepin. Data were calculated by a graphical analysis on voxel-by-voxel and ROI (region of interests) basis. Binding potential (BP) values for [(11)C]-doxepin binding in the frontal and prefrontal cortices, and cingulate gyrus were significantly lower in the depressed patients than those in the normal control subjects. There was no area of the brain where [(11)C]-doxepin binding was significantly higher in the depressed patients than in the controls. ROI-based analysis also revealed that BP values for [(11)C]-doxepin binding in the frontal cortex and cingulate gyrus decreased in proportion to self-rating depressive scales scores. The results of this study demonstrate that depressed patients have decreased brain H(1)R binding and that this decrease correlates with the severity of depression symptoms. It is therefore suggested that the histaminergic neuron system plays an important role in the pathophysiology of depression and that its modulation may prove to be useful in the treatment of depression.     

Binding of [3H]histamine to receptor sites in rat brain.

[³H]Histamine binds to crude synaptic membranes from rat brain with a dissociation constant of 0.683 μm. The hypothalamus shows the most binding among five other brain regions. Binding of [³H]histamine is not affected by noradrenaline, dopamine, or serotonin; however, it is inhibited more by H₂ receptor than by H₁ receptor agents, suggesting the prevalence of H₂-histamine receptors in brain.     

Comparison of the size of neuronal and non-neuronal histamine pools in the brain of different rat strains

Summary The size of the neuronal and non-neuronal histamine pools in the brain of three different strains of rats was measured by assuming that the -fluoromethylhistidine-induced maximal decrement of histamine represents the size of the neuronal pool. Although the total histamine levels in the brain showed a considerable interstrain variation, no significant interstrain difference was observed in the neuronal histamine level. These results suggest that the size of the neuronal histamine pool in the brain is relatively stable, whereas the size of the non-neuronal histamine pool is variable.     

Circadian rhythm photic phase shifts are not altered by histamine receptor antagonists

Histamine may play a role in synchronizing endogenous circadian rhythms with exogenous photic cues. Direct application of histamine to the suprachiasmatic nucleus, the site of the mammalian circadian pacemaker, phase shifts the circadian rhythm in neural activity [7]. Intraventricular injections of histamine also phase shift circadian rhythms [14]. The magnitude and direction of the phase shifting effects of histamine depend on circadian phase in a manner similar to light [7,14]. Depletion of brain histamine levels by inhibition of histamine synthesis reduces phase shifts to light [10].     

Regeneration of ascending spinal axons in goldfish

Regeneration of descending spinal cord tracts occur spontaneously in adult goldfish. Very little information is available regarding the fate of ascending fibers. Using Dextran amines as a tracer, we studied the normal and regenerated ascending axonal projection patterns in adult goldfish brain nuclei. Present study includes spinal projections to torus semicircularis, hypothalamus, thalamus and the telencephalon. Regenerated fibers had finer caliber axons and the terminal axonal arbors covered a larger area than the corresponding normal ones.     

Circadian rhythms in spontaneous neuronal discharges of the cultured suprachiasmatic nucleus

The suprachiasmatic nucleus (SCN) is believed to play a major role in the generation and control of circadian rhythms in mammals. In order to obtain further evidence concerning this, single and multiple neuronal discharges were continuously recorded over a period of several days in neonatal rat SCN explants. These organotypic explants, which had been cultured for several weeks in a chemically defined medium, showed alternating high and low levels of spontaneous neuronal discharges with a periodicity around 24 h. Such explants can serve as a useful model to study the neuronal mechanisms underlying the generation of mammalian circadian rhythms.     

Circadian variation of serum thyrotropin in endogenous depression

The circadian variation of serum thyrotropin (thyroid-stimulating hormone; TSH) was studied in nine patients with endogenous depression before and after recovery. Depressed state did not appear to influence the pattern of TSH. When 2 mg of dexamethasone was administered, serum TSH was significantly reduced for 18 hours, whereafter the effect leveled off. The TSH response to thyrotropin-releasing hormone (TRH) was evaluated 25 hours after the administration of dexamethasone and the response was found to be unchanged.     

Decrease in histamine turnover in the brain of spontaneously hypertensive rats

Histamine (HA) turnover rate in the brain of spontaneously hypertensive rats (SHR) was determined by the accumulation of telemethylhistamine after pargyline treatment. The values in these SHR were lower than in the Wistar Kyoto rats, particularly in the hypothalamus and brainstem. However, chronic treatment with L-histidine had no effect on the development of hypertension in the SHR. The functional significance of the decreased HA turnover in SHR is discussed in relation to the pathogenesis of hypertension.