5-Hydroxytryptophan (5-HTP)-immunoreactive neurons in the rat brain tissue

We demonstrated the presence of 5-hydroxytryptophan (5-HTP), the immediate precursor of serotonin (5-HT), in the rat brain tissue using a glutaraldehyde-coupled immunohistochemical technique. The immunoreactivity of 5-HTP was intensified in the colchicine-pretreated rat. The distribution of labelled cells was the same as for 5-HT-immunoreactive cells, but they were fewer in number.     

Voltammetric detection of the release of 5-hydroxyindole compounds throughout the sleep-waking cycle of the rat

Summary In the present work, differential pulse voltammetry (DPV) measurements of the extracellular fraction of 5-hydroxyindole compounds were performed in rats under long-term chronic conditions. In the nucleus Raphe Dorsalis (n.RD), the voltammetric signal measured at +300 mv (peak 3) disappeared completely 70 to 90 min after injection of Clorgyline (10 mg/kg), a monoamine oxidase inhibitor type A (MAOI-A); the signal measured in such conditions is thus dependent upon extracellular 5-hydroxyindoleacetic acid (5-HIAA peak 3). Deprenyl, an MAOI type B, at the same dose, induced only a slight increase in peak 3 height; according to the fact that MAO-B is selectively located in the 5-HT neurons and since their inhibition does not decrease 5-HIAA peak 3 nor the endogenous 5-HIAA content as measured with High Performance Liquid Chromatography (HPLC), 5-HIAA measured with DPV in the extracellular fluid of untreated animals might come from 5HT released and metabolized by MAO-A outside the 5-HT neurons. In animals implanted for measurements of both voltammetric and polygraphic parameters, the 5-HIAA peak 3 measured mainly in the anterior and ventral part of the n.RD exhibited large increases in its height during slow-wave sleep (SWS: +39%) and paradoxical sleep (PS=+71%) as compared to the waking state (W=100%); these variations could reflect the dendritic release of 5-HT. In the Caudate nucleus (n.Cd) the same voltammetric signal presented reverse fluctuations, i.e. an increase during W and a decrease during SWS and PS. Intracerebroventricular administration of Corticotropin-Like Intermediate lobe Peptide (CLIP, 10 ng/2 l) induced an increase in PS duration (+51%) preceded and accompanied by an increase in the n.RD 5-HIAA peak 3 height (+50%).     

Detection of the release of 5-hydroxyindole compounds in the hypothalamus and the n. raphe dorsalis throughout the sleep-waking cycle and during stressful situations in the rat: a polygraphic and voltammetric approach

Summary In the present work, voltammetric method combined with polygraphic recordings were used in animals under long-term chronic conditions; the extracellular concentrations of 5-hydroxyindole compounds (5-OH^les) and in particular 5-hydroxyindoleacetic acid (5-HIAA) were measured in the hypothalamus and in the nucleus Raphe Dorsalis (n.RD). The hypothesis that extracellular detection of 5-HIAA, in animals under physiological conditions, might reflect serotonin (5-HT) release is suggested by the following observations: — serotoninergic neurons are reported to contain only monoamine oxidase type B (MAO-B); — an inhibitor of such an enzyme, MDL 72145 (1 mg/kg), fails to decrease the extracellular 5-HIAA peak 3 height; — MAO type A is contained in non-5-HT cells or neurons; — only the inhibitor of this last type of enzyme (Clorgyline 2.5 mg/kg) induces a complete disappearance of the voltammetric signal. The 5-HIAA measured in the extracellular space thus comes from the 5-HT released and metabolized outside the 5-HT neurons. Throughout the sleep-waking cycle, 5-OH^les release occurs following two different modes: 1 — during sleep, in the vicinity of the 5-HT cellular bodies in the n.RD; this release might come from dendrites and be responsible for the 5-HT neuronal inhibition occurring during sleep; 2 — during waking, at the level of the axonal nerve endings impinging on the hypothalamus; this release might be related to the synthesis of hypnogenic factors. Finally, we have observed that in the hypothalamus, 30 min. of immobilization-stress (IS) induces a larger increase of the voltammetric signal (+ 80%) than a painful stimulation of the same duration (+ 30%); the possible link between the 5-OH^les release occurring in this area during an IS and the subsequent paradoxical sleep rebound is discussed.     

The contribution of sleep to improvements in working memory scanning speed: a study of prolonged sleep restriction

Working memory scanning and motor response speeds were assessed in chronically sleep restricted participants using the Sternberg item recognition paradigm (SIRP). Twenty-two healthy volunteers (ages 21-30) living in a controlled hospital environment were allowed either 4h of sleep opportunity (50% of habitual sleep) or 8h of sleep opportunity (100% of habitual sleep) for 12 days. Working memory scanning efficiency (time taken to access an item in working memory) was tested for the first 9 days of sleep restriction and improved over time in participants permitted an 8h sleep period, but did not change significantly in participants permitted a 4h sleep period. Speed of motor response (reaction time independent of cognitive processing) did not change significantly in either group. These results indicate that the efficiency of working memory scanning can improve with repeated practice given sufficient sleep, and that prolonged sleep restriction to 50% of habitual sleep prevents this improvement.     

Binocular and monocular/unihemispheric sleep in the domestic chick (<Emphasis Type="Italic">Gallus gallus</Emphasis>) after a moderate sleep deprivation

Binocular (Bin-sleep) and monocular/unihemispheric sleep (Mo-Un sleep) were studied in domestic chicks (Gallus gallus) after an 8 h period of sleep deprivation. Eleven-day-old chicks were divided into three groups: two non-deprived (N-DEP1 and N-DEP2) and one deprived for 8 h (DEP-8H). Deprivation was performed by placing chicks on a treadmill on which they were forced to walk continuously. Sleep behaviour was recorded for 6 h consecutively immediately after the end of sleep deprivation. During the recovery period, sleep-deprived chicks slept for a longer duration, spent significantly more time in binocular sleep and slept for significantly longer episodes that did control chicks. Regarding Mo-Un sleep, sleep deprivation seems to affect the right hemisphere by reducing the number of episodes and the time spent sleeping with the left eye closed. These results suggest that sleep-deprivation significantly influences the pattern of sleep in domestic chicks allowing for a better recovery. The authors state that they have adhered to the “Principles of laboratory animal care” (NIH publication N° 86-23, revised 1985) and to the legal requirements of our country during the present study.     

Reorganization in the auditory cortex of the rat induced by intracortical microstimulation: a multiple single-unit study

Many manipulations are able to change or perturb various aspects of single neuron properties and interneuronal relationships. Changes of cerebral cortex organization have been observed in different cortical areas and at different time scales in relation to peripheral stimulation, peripheral damage, associative learning, and electrical stimulation. Here we describe studies on separable multineuron recordings in the rat's auditory cortex under two different anesthetics. Acoustic stimuli were used as a normal, physiological input, and weak electrical intracortical microstimulation (ICMS) as a perturbation that forces a rapid cortical reorganization. ICMS induced fast changes in the cortical map and in the receptive field properties of cells at the electrically stimulated and adjacent electrodes. In effect there was an enlargement of the cortical domain tuned to the acoustic frequency that had been represented at the stimulating electrode. ICMS also incremented afterdischarge responses; these consisted of an initial response to the auditory stimulus followed by less intense repetitive activity that was stimulus-time locked and had a period of 8–12 Hz, similar to that of the spontaneous synchronous activity. Cortical activity under ketamine differed from that under pentobarbital sodium, although in both situations we observed that cortical neurons were highly synchronous.     

The role of growth/differentiation factor 5 (GDF5) in the induction and survival of midbrain dopaminergic neurones: relevance to Parkinson's disease treatment

Growth/differentiation factor-5 (GDF5) is a member of the transforming growth factor-beta superfamily which has potent effects on dopaminergic neurones in vitro and in vivo. GDF5 is under investigation as a potential therapeutic agent for Parkinson's disease (PD), which is caused by the progressive degeneration of dopaminergic neurones projecting from the substantia nigra (SN) to the striatum. In the rat ventral mesencephalon (VM; the developing SN), GDF5 expression peaks at embryonic day 14, the time at which dopaminergic neurones undergo terminal differentiation. Addition of GDF5 protein to cultures of embryonic rat VM increases the survival and improves the morphology of dopaminergic neurones in these cultures. GDF5 treatment also increases the number of cells which adopt a dopaminergic phenotype in cultures of VM progenitor cells. Intracerebral administration of GDF5 has potent neuroprotective and restorative effects on the nigrostriatal pathway in animal models of PD. Furthermore, addition of GDF5 protein to embryonic rat dopaminergic neuronal transplants improves their survival and function in a rat model of PD. Thus, GDF5 has potential applications to PD therapy as a dopaminergic neuroprotective agent and as a factor that may induce a dopaminergic neuronal fate in unrestricted progenitor cells.     

The role of 5-HT receptor subtypes in the ventrolateral orbital cortex of 5-HT-induced antinociception in the rat

The present study examined the involvement of 5-HT in the ventrolateral orbital cortex (VLO) on descending antinociception and determined which subtypes of 5-HT receptors mediated this effect. This study focused on the effects of 5-HT microinjection in the VLO of lightly anesthetized male rats on the radiant heat-evoked tail flick (TF) reflex, as well as the influence of 5-HT(1A), 5-HT(2), 5-HT(3), and 5-HT(4) receptor subtype antagonists on the effect of 5-HT. Results showed that 5-HT microinjection (2, 5, 10 microg, in 0.5 microl) into the VLO depressed the TF reflex in a dose-dependent manner. Pretreatment with 5-HT receptor antagonists (1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl] piperazine hydrobromide (NAN-190), cyproheptadine hydrochloride (CPT) and 1-methyl-N-(8-methyl-8-azabicyclo[3.2.3]-oct-3-yl)-1H-indazole-3-carboxamide maleate salt (LY-278,584)), specific for 5-HT(1A), 5-HT(2) and 5-HT(3) receptors, respectively, partially reversed the 5-HT-evoked inhibition. In contrast, the 5-HT(4) receptor antagonist, 1-[2-[(methylsulfonyl)-amino]ethyl]-4-piperidinyl]methyl1-methyl-1H-indole-3-carboxylate (GR 113808), had no effect on the inhibition of 5-HT. Microinjections of NAN-190, CPT and LY-278,584 alone into the VLO had no effect on the TF reflex. These results suggest that 5-HT(1A), 5-HT(2) and 5-HT(3), but not 5-HT(4) receptors, are involved in mediating 5-HT-induced antinociception in the VLO. According to different properties and distribution patterns of the 5-HT receptor subtypes on neurons, the possible mechanism of 5-HT activation of the VLO-periaqueductal gray (PAG) descending antinociceptive pathway is discussed.     

Urinary excretion of 5-hydroxyindoleacetic acid (5-HIAA) in the rat after immobilization stress

An increase of 100% in urinary 5-hydroxyindoleacetic acid, as a measure of release and catabolism of serotonin, is associated with the stress of 18 hr of restraint. Seventy percent of the rats developed gastric ulcers. Increased urinary 5-hydroxyindoleacetic acid levels were not observed in rats deprived of food for 42 hr.     

The light/dark difference in meal size in the laboratory rat on a standard diet is abolished during REM sleep deprivation

Using a modified flowerpot technique, by which it was possible to use test animals as their own controls, the meal patterns of eight adult male rats were recorded before, during and after rapid eye movement sleep (REMs) deprivation. The results demonstrated that the prominent light/dark (LD) difference in meal size, typical of the baseline meal pattern, was abolished during REMs deprivation. This equalization was entirely due to reduction in meal size during the dark hours. After termination of REMs deprivation the size of the dark meals increased almost to the baseline level. However, the LD difference was not restored since the size of the light meals was also significantly increased. Changes in meal frequency were confined to the period of REMs deprivation. The possible role of REMs as a modulator of meal size in relation to the LD cycle of illumination is discussed.     

The role of 5-methoxytryptophan in pediatric-onset lupus nephritis: A retrospective cohort study

BackgroundThis clinical study investigates the role of 5-methoxytryptophan (5-MTP) in pediatric systemic lupus erythematosus (SLE), with a particular interest in lupus nephritis (LN).Patients and methodsOne hundred ten children with SLE were enrolled in the cohort study. Among the patients, seventy-seven (70%) had active LN and thirty-three (30%) were not present with LN during their first visit to the clinic. The diagnoses of LN were biopsy-proven. Serum samples were collected before and after administration of immunosuppressive medications to evaluate 5-MTP levels and regular laboratory data. Data were analyzed longitudinally.ResultsBefore any treatment started, patients with active LN had significantly higher 5-MTP levels as compared to patients with no LN (1.021 ± 0.709 vs. 0.719 ± 0.606, P = 0.0456). Also, in patient with active LN, 5-MTP level was significant decreased after treatment, compared with the levels before treatment (1.021 ± 0.709 vs. 0.802 ± 0.597, P = 0.0484). Patients who reached complete remission also had significantly higher initial serum 5-MTP levels than that in patients with no remission (1.244 ± 0.784 vs. 0.846 ± 0.556, P = 0.0488). There was an overall reduction in 5-MTP levels after six months of immunosuppressive treatment, regardless of the disease outcome. Subgroup analysis further revealed a significantly higher 5-MTP level during the active stage of LN (1.127 ± 0.149 vs. 0.742 ± 0.092, P = 0.0384).ConclusionWe demonstrated that serum 5-MTP level is positively correlated to the disease activity, prognosis, and remission status of pediatric LN in vivo.     

C5a anaphylatoxin as a product of complement activation up-regulates the complement inhibitory factor H in rat Kupffer cells

The 155-kDa complement regulator factor H (FH) is the predominant soluble regulatory protein of the complement system. It acts as a cofactor for the factor I-mediated conversion of the component C3b to iC3b, competes with factor B for a binding site on C3b and C3(H2O) and promotes the dissociation of the C3bBb complex. The primary site of synthesis is the liver, i.e. FH-specific mRNA and protein were identified in both hepatocytes (HC) and Kupffer cells (KC). Previous studies in rat primary HC and KC had shown that the proinflammatory cytokine IFN-gamma influences the balance between activation and inhibition of the complement system through up-regulation of the inhibitory FH. In this study we show that C5a, as a product of complement activation, stimulates the expression of FH-specific mRNA and protein in KC and thus induces a negative feedback. Quantitative-competitive RT-PCR showed an approximate threefold C5a-induced up-regulation of FH. ELISA analyses revealed a corresponding increase in FH protein in the supernatants of KC. The up-regulation of FH was completely inhibited by the C5a-blocking monoclonal antibody 6-9F. Furthermore, an involvement of LPS and IFN-gamma was excluded, which strongly indicates a direct effect of C5a on the expression of FH in KC.     

The role of CD5 expression in thymic carcinoma: possible mechanism for interaction with CD5+ lymphoid stroma (microenvironment)

Recently, an increasing number of TFE3 rearrangement‐associated tumours have been reported, such as TFE3 rearrangement‐associated perivascular epithelioid cell tumours (PEComas), melanotic Xp11 translocation renal cancers and melanotic Xp11 neoplasms. We have suggested that these tumours belong to a single clinicopathological spectrum. ‘Xp11 neoplasm with melanocytic differentiation’ or ‘melanotic Xp11 neoplasm’ have been proposed to designate this unique neoplasm. Herein, we describe the first case of an Xp11 neoplasm with melanocytic differentiation to be described in the prostate, bearing the novel NONO–TFE3 gene fusion. This study both adds to the spectrum regarding melanotic Xp11 neoplasms and expands its gene fusion spectrum. Moreover, we discuss the relationship of these rare tumours to neoplasms such as conventional PEComas, alveolar soft part sarcomas, malignant melanomas, clear cell sarcomas and Xp11 translocation renal cancers.     

Intra-ventricular infusion of the NMDA antagonist AP5 impairs performance on a non-spatial operant DRL task in the rat

Summary Rats were trained to lever press on a differential reinforcement of low rates (DRL-18 s) schedule. They were then allocated to four treatment groups. These were: hippocampal aspiration lesions [HIPP]; implantation of osmotic minipumps for intraventricular infusion of either (a) the NMDA receptor antagonist 30 mM D, L-2-amino-5-phosphonopentanoic acid [AP5] or (b) vehicle [VEH]; and an unoperated control group [UNOP]. In subsequent DRL testing, the HIPP group showed a profound and enduring loss of efficiency, resulting from an increased tendency to respond too early; the AP5 group showed a qualitatively similar, but less severe, impairment followed by full recovery once the minipumps had expired; the VEH and UNOP groups both maintained their pre-operative levels of efficiency. We conclude that AP5 infusion disrupts temporary memory storage in the hippocampus, and that the hippocampus is concerned with the retention of memories outside the purely spatial domain.     

The ontogenetic development of serotonin (5-HT1) receptors in various cortical regions of the rat brain

Summary The distribution of serotonin (5-HT₁) receptors in various cortical regions of the rat brain has been examined during ontogenesis by quantitative autoradiography.An increase in binding site density between the first postnatal day and adult age was observed and could be approximated by a sigmoid shaped (logistic) growth curve. A marked heterochrony in the increase of binding site density is found in the 13 analyzed cortical regions. Binding sites develop earlier in neocortex than in allocortical areas. Fifty pereent of the binding site density of adult age is reached in the motor cortex at the 9th postnatal day, followed by the primary somatosensory cortex one day later, by the medial prefrontal cortex on the 12th day, by the fascia dentata on the 14th day and by the CA1-region on the 20th day. A detailed analysis of the frontal, medial prefrontal and hippocampal regions also shows a heterochrony within these regions. Adult values of binding site densities are also reached at different ages in the various cortical regions. The highest receptor densities were observed in the dorsal subiculum, the lowest in the primary somatosensory cortex.     

蛋白激酶C和激活蛋白-1信号级联对哮喘大鼠IL-5基因转录的调控作用

目的　探讨蛋白激酶C(PKC)和激活蛋白 -1(AP- 1)信号转导级联在支气管哮喘(简称哮喘)大鼠外周血T淋巴细胞白细胞介素5 (IL -5 )基因转录中的作用。方法　建立大鼠哮喘模型。从每只大鼠外周血中分离T淋巴细胞,并随机分为空白对照组、PKC激动剂12 肉豆蔻酰 13 乙酸佛波酯(PMA)组、PMA和AP -1顺式诱骗元件(CDODNs)组及PMA和AP -1突变顺式诱骗元件(突变CDODNs)组进行培养。两种CDODNs分别经脂质体转染至后两组细胞。用电泳迁移率改变分析(EMSA)检测AP -1活化,用逆转录聚合酶链反应(RT- PCR)检测IL -5mRNA表达。结果　(1)PMA组的AP -1活化(86 777.2 2±2 2 5 7.79)和IL 5mRNA表达(0 .810 0±0 .0 316 )增加,与空白对照组(分别为2 0 70 8.5 4±96 8.0 4和0 .30 5 0±0 .0 12 9)相比较,差异有统计学意义(P <0 .0 1)。(2 )PMA和AP -1顺式诱骗元件组的AP -1活化(2 3475 .90±75 7.99)被抑制,IL -5mRNA表达(0 .345 0±0 .0 12 9)亦减少,与PMA组相比较,其差异有统计学意义(P <0 .0 1)。PMA和AP 1突变顺式诱骗元件组的AP 1活化(86 95 5 .71±16 0 0 .38)和IL- 5mRNA表达(0 .8175±0 .0 2 2 2 )与PMA组相比较,差异无统计学意义(P >0 .0 5 )。(3)T淋巴细胞AP -1活化和IL 5mRNA表达呈显著正相关(P <0 .0 1)。结论　哮     

The role of ARMC5 in adrenal pathology: a brief review

Recent advances in molecular analysis in the past decade have enabled the identification of genes associated with endocrine disease states. ARMC5 is among those genes identified, one which pathologists should know for the purposes of directing care in those patients harboring these variants. This review discusses the role of ARMC5 in adrenal gland pathology, focusing on primary bilateral adrenal cortical macronodular hyperplasia and, specifically, what is known about this disease at this point in time.     

The effect of the timing, quality and quantity of sleep upon the depression (masking) of body temperature on an irregular sleep/wake schedule

SUMMARY  Eight subjects were studied on an irregular sleep/wake schedule which was designed so that prior wake time and the time of day when 4-h sleep periods were taken were both balanced. Rectal temperature and the sleep EEG were measured throughout the 8-d protocol. A purification method was used to estimate the depression of rectal temperature (masking) caused by the first and second halves of each 4-h sleep period. On average this was 0.187 ± 0.015°C during the first 2h and 0.262 ± 0.017°C during the second 2h. Masking increases beyond the first 2 h of sleep, but any effects due to the phase of the rectal temperature when sleep is taken are very weak. However, masking did increase (that is, the temperature was depressed more) when the amount of prior wake time was greater than 4 h. When the effects of sleep content variables were considered also, masking was still greater in the second half of sleep and tended to increase with the amount of slow-wave sleep, but it decreased with increasing amounts of time awake during a sleep period (sleep latency or sleep discontinuity). Some implications of these results for the mechanism of sleep-induced masking are considered.