The effect of MPTP on the neuronal uptake of monoamines]

The results of kinetic analysis of synaptosomal uptake of dopamine, noradrenaline, adrenaline and serotonin showed the presence of their own carrier systems with high or low affinity for each monoamine. The low affinity system of the uptake of monoamines by nerve endings differs from extraneuronal one by higher affinity. MPTP noncompetitively inhibits the system of highly effective uptake of the studied monoamines by nerve endings, competitively inhibiting synaptosomal uptake with low affinity of noradrenaline, adrenaline and noncompetitively serotonin and dopamine. The constant values of inhibition showed that MPTP most strongly blocks the system of synaptosomal uptake of low affinity serotonin and approximately 2-times weaker affects its system of high affinity. Carrier systems of high affinity of dopamine, adrenaline and noradrenaline block MPTP 150-500 times weaker than that of serotonin, and as for low affinity--in 2000-4000 times. It may be supposed that synaptosomal uptake of low affinity serotonin is most perceptible to the effect of MPTP and is of a particular importance in the development of Parkinson's disease symptoms.     

Catecholamines and indolalkylamines of the pia mater and spinal cord in vertebrates

Fluorimetric methods were used to determine adrenaline, dopamine, noradrenaline, serotonin and tryptamine in the pia mater of the brain and spinal cord of various vertebrates (fishes, birds, mammals) and of man. The histochemical method using glyoxylic acid showed the presence of biogenic monoamines in adrenergic nerve fibres and in the monoaminocytes. Their total amount in the pia mater is roughly the same, except in man, in whom it is significantly lower. From the higher adrenergic axon concentration on the one hand and the lower number of monoaminocytes on the other, it can be concluded that the neuronal factor has a more important role in the regulation of brain haemodynamics in man.     

Changes in the balance of biogenic monoamines and their metabolites in the organs of rats with oxygen-induced epilepsy

The content of some biogenic monoamines and their metabolites in rat brain and heart in different periods of oxygen epilepsia was studied using high performance liquid chromatography with electrochemical detection. It was shown that already at the 5th minute of exposure to oxygen adrenaline, DOPA and some noradrenaline metabolites disappeared in the brain and noradrenaline level reduced. At this period in rat heart the reduction of catecholamine content was the most distinct and serotonin level was unchanged. At the beginning of convulsive period the modifications of biogenic amines content were nonparallel in brain regions: in the heart the reduction of catecholamine level went on, especially in right ventricle. In the terminal phase of oxygen epilepsia brain biogenic amines increased, however, not up to normal meaning, heart catecholamines at this period were at the same level as at the beginning of the convulsive period.     

7株野生虫草菌的鉴定及其菌丝体醇提取物对HepG2细胞的抑制活性

结合形态学与ITS序列分析对7株野生虫草真菌进行分类鉴定。MTT法分析它们的菌丝体醇提取物对肝癌HepG2细胞增殖的抑制活性。鉴定结果表明菌株MF7、MF9、MF14为细脚棒束孢Isaria tenuipes,菌株MF11、MF12、MF13为蝉棒束孢Isaria cicadae,菌株MF10为球孢白僵菌Beauveria bassiana;MTT结果显示分离到的3株细脚棒束孢和3株蝉棒束孢的菌丝体醇提取物对HepG2的抑制活性较差,IC₅₀均大于500μg/mL;球孢白僵菌MF10对HepG2细胞有一定抑制作用,IC₅₀值为221.6μg/mL,略强于蝙蝠蛾拟青霉发酵菌丝粉产品金水宝胶囊（IC₅₀=364μg/mL）和中华被毛孢发酵菌丝粉产品百令胶囊（IC₅₀=268.7μg/mL）。另外,发现供对比试验的3株蛹虫草菌株（MF1、MF5、MF15）对HepG2细胞均有较好的抑制作用,其中MF15的发酵菌丝体醇提取物活性最强,IC₅₀为55.56μg/mL,暗示蛹虫草发酵菌丝体具有重要的研究价值。     

The effects of biogenic monoamines and related agonists and antagonists on the isolated,perfused branchial heart of sepia officinalis L. (cephalopoda)

1. The chronotropic and inotropic effects of biogenic monoamines were examined on the isolated and perfused branchial heart of the common cuttlefish Sepia officinalis (L.).2. Adrenaline, noradrenaline and dopamine caused concentration-dependent increases in pressure amplitude, dopamine being 100-fold less potent than adrenaline and noradrenaline. The catecholamines hardly affected frequency.3. Octopamine, histamine and GABA did not influence normal heartbeat.4. Serotonin was either ineffective or produced variable responses, whereby the efficacy of the substance was not graded to the concentrations applied. It is supposed that serotonin is not involved in branchial heart regulation.5. The actions of different adrenergic agonists and antagonists indicate the presence of a myocardial adrenoceptor which closely resembles the α₁-type.     

Biogenic Amines in the Vomeronasal Organ

The vomeronasal organ of frog and mouse was investigated forthe presence and content of serotonin and catecholamines bymeans of high-performance liquid chromatography. Measurableamounts of serotonin, adrenaline and noradrenaline were foundin the vomeronasal organ of adult individuals of both species.The amine content varied with sex of adult frogs and mice andsexual maturity of mice. In preliminary experiments, acute exposureto male urine containing pheromone affected the amine contentin the vomeronasal organ of adult female mice. These data suggestthat functional sex dimorphism is present in the vomeronasalorgan, and biochemical changes therein take place accordingto stage of sexual maturity. The role of biogenic amines inthe vomeronasal organ deserves further study. Chem. Senses 22:439–445, 1997.     

Adrenergic innervation of the oesophagus in the cat (Fellis domestica) and rhesus monkey (Macacus rhesus)

Summary The distribution of monoamines in the pharynx and oesophagus of the rhesus monkey (Macacus rhesus) and the cat (Felis domestica) was investigated by means of fluorescence microscopical and chemical methods. Fluorimetric determinations reveal the presence of varying amounts of noradrenaline in the pharynx and oesophagus of the rhesus monkey. The lowest amount (0.05 (g/g) was found in the lower part of the oesophagus, the so-called sphincter-segment. The middle and upper part of the oesophagus contain medium amounts of noradrenaline (0.06–0.09 g) whereas the highest concentration was detected in the pharynx (0.14 (g/g). Neither dopamine nor adrenaline occurred in the tissue pieces analyzed. Fluorescence microscopically noradrenaline was found to be located in varicose intramural nerve fibre plexus which innervate mucous glands and blood vessels in the pharynx of both species. In the rhesus monkey, the lamina muscularis mucosae of all parts of the oesophagus is supplied by a well developed noradrenergic ground-plexus. Preterminal and terminal varicose nerve fibres are distributed in myenteric and submucous ganglia of the oesophagus; the number of such ganglia decreases towards the lower segment. The density of the adrenergic innervation is higher in myenteric when compared to submucous ganglia. The arrangement of the intraganglionic terminals suggests that both axosomatic and axodendritic contacts occur in Auerbach's ganglia whereas axodendritic contacts seem to predominate in Meissner's ganglia. Myenteric ganglia situated close to the submucosa as well as true submucous ganglia may be occasionally seen to be traversed by faintly fluorescent non-varicosed fibres which do not establish any synaptic contacts. The fluorescence intensity of intraganglionic varicosities varies considerably; accordingly the transmitter content of individual varicosities seems to be very variable. The adrenergic innervation of the lamina muscularis is restricted to single contorted fibres being sparsely distributed throughout the longitudinal smooth muscle layer. The circularly arranged smooth musculature of the sphincter-segment lacks an adrenergic nerve supply. The vagus nerve carries sympathetic adrenergic fibres to the lower oesophagus and the cardia. Species differences between the innervation pattern in rhesus monkeys and cats are outlined: No adrenergically innervated ganglia occur in the submucosa of the cat. However, part of the myenteric ganglia in cats exhibit an adrenergic innervation pattern similar to that seen in submucous ganglia of the rhesus monkey. They might therefore be regarded as morphologically equivalent to the plexus submucosus which is, however, present in the whole gut. The density of the noradrenergic ground-plexus in the muscularis mucosae of the cat's oesophagus is less than that of the corresponding plexus in rhesus monkeys.The influence of noradrenaline upon the smooth musculature and the neurons from myenteric as well as submucous ganglia is discussed. From the point of view of the adrenergic innervation there is no structure corresponding to the sphincterlike lower oesophageal segment.Supported by the Deutsche Forschungsgemeinschaft and the Joachim-Jungius-Gesellschaft zur Förderung der Wissenschaften, Hamburg.     

Identification and quantitative measurements of biogenic monoamines in the central nervous tissue of some gastropods

Investigations were carried out to determine levels of biogenic monoamines in the central nervous tissue of seven species of terrestrial gastropods (taxonomic family: Helicidae) using high-performance liquid chromatography (HPLC) with electrochemical detection (ELCD). The central nervous tissue of all species examined contained assayable amounts of dopamine (DA) and serotonin (5-HT). Noradrenaline (NA) was occasionally present; adrenaline (A) could not be detected. The central ganglia of two Mediterranean species generally contain 3-4 times more 5-HT relative to DA than the ganglia of the other species which contain amounts of 5-HT only slightly higher than DA.     

Maackiain in Cicer bijugum Rech. f. associated with resistance to Botrytis grey mould

Resistance in Cicer bijugum Rech. f. a wild relative of chickpea, to Botrytis grey mould (BGM), caused by Botrytis cinerea Pers., was shown to be associated with high concentrations of maackiain when compared to three susceptible species. The two BGM resistant accessions of C. bijugum contained between 200 and 300 μg maackiain g⁻¹ of foliage whereas the BGM susceptible species C. arietinum, C. echinospermum and C. reticulatum contained less than 70 μg g⁻¹. Furthermore, the concentration of maackiain increased to more than 400 μg g⁻¹ in the resistant wild species after being inoculated with the pathogen whereas no significant increase was recorded in the susceptible species. The germination of spores of Botrytis cinerea, treated with maackiain, was inhibited in a dose dependent manner; less than 10% of spores germinated when treated with 500 μg ml⁻¹. The data indicate that maackiain may be an important component in BGM resistance in the wild chickpea C. bijugum and that the resistance is enhanced in the presence of the pathogen.     

The effect of biogenic monoamine antagonists on the development of preimplantation mouse embryos cultured in vitro

The effect of serotonin and adrenaline antagonists was tested on the early embryos of mice of three lines. All the substances tested produced an arrest or inhibition of cleavage division and the appearance of anomalies. Serotonin introduced in the incubation medium was effective against some serotoninolytics. We were unable to test the protective effect of adrenaline, as in the concentrations used it has its own effect on the development. From the data obtained, a conclusion is made of the existence in early mouse embryos of the structures sensitive to serotonin and adrenaline antagonists. The assumptions is made from the previously obtained data on the presence of biogenic monoamines in early mouse embryos, of functional activity of prospective mediators of the nervous system at the earliest stages of embryonic development of mammals.     

蝙蝠蛾幼虫肠道中的冬虫夏草菌体研究

西藏林芝地区冬虫夏草天然产区野生虫草蝙蝠蛾幼虫的肠道液pH范围为9.69-10.99,体外在此pH下培养冬虫夏草菌长势较差。同时,在该pH的体外模拟中肠液中,虫菌体也会在极短的时间内消亡,而对自然侵染的蝙蝠蛾幼虫的血淋巴、消化道及其“反吐物”（肠液）进行切片和显微镜检却发现,蝙蝠蛾幼虫消化道、呕吐物中均存在与血淋巴中相同的冬虫夏草菌菌体形态,生存状态良好,正常增殖,表明蝙蝠蛾幼虫肠道的生理状态可以维系并支撑冬虫夏草菌菌体某种形态的存在与生长,并证实冬虫夏草菌由口器及消化道入侵蝙蝠蛾幼虫的可能性;肠道虫菌体与血淋巴虫菌体不一定同时存在,表明冬虫夏草菌对蝙蝠蛾幼虫的侵染可能同时存在表皮侵染和肠道侵染两种途径。本文为冬虫夏草菌对蝙蝠蛾幼虫的侵染途径研究提供更加多样化的信息支持。     

Computer-operated microspectrofluorimetry to identify formaldehyde-induced fluorophores of biogenic monoamines and precursor substances in models and tissue sections

By means of a histochemical reaction using formaldehyde vapour (Falck and Owman 1965), biogenic monoamines and precursor substances, i.e., L-DOPA, dopamine, noradrenaline, adrenaline, 5-hydroxytryptophan and 5-hydroxytryptamine, may be converted into fluorophores with specific spectral characteristics, i.e., the emission spectrum, excitation spectrum and fading curve. The registration and correction of the spectral properties and changes induced by acidification with hydrochloric acid vapour or treatment with ammonia vapour, of these formaldehyde-induced fluorophores, are performed by an automated microspectrofluorimeter, developed by modification of a Leitz MPV 2 system. This work deals with the instrumental configuration and certain methodological features in order to identify the fluorogenic biogenic monoamines and precursor substances in models and tissue sections. Registrations of excitation peak values, for the first time extended to a wavelength range from 240-460 nm, are discussed, which enable the calculation of peak ratio values 410/260, 380/320, 320/260 or 385/315, suitable as identification parameters for formaldehyde-induced fluorophores of biogenic monoamines and precursor amino acids.     

Biogenic amine regulation of bovine luteal progesterone production in vivo

Biogenic amines were administered using osmotic pumps placed subcutaneously in the neck region of regularly cycling, non-lactating dairy cows on Days 9-11 (oestrus = Day 0) of the oestrous cycle. Blood samples were collected using indwelling jugular catheters and the plasma progesterone concentrations were measured. Samples were collected at 4-h intervals for the first 12 h of treatment and thereafter at 12-h intervals for the remainder of the 72-h treatment period. After administration of various doses of noradrenaline, adrenaline and serotonin (0.5-2.0 micrograms/kg/h) significant elevation of plasma progesterone was achieved at a dosage of 2.0 micrograms/kg/h (P less than 0.01). The response to adrenaline was greater than that observed for noradrenaline and serotonin (P less than 0.05). Within-treatment comparison to pretreatment samples showed plasma progesterone concentrations to increase within 4 h after the administration of noradrenaline, adrenaline and serotonin (P less than 0.05) and this enhancement was maintained throughout the treatment period (P less than 0.05). The elevation in plasma progesterone concentrations induced by noradrenaline, adrenaline and serotonin was independent of changes in circulating concentrations of luteinizing hormone. These results support a physiological role for endogenous biogenic amines in the control of bovine luteal progesterone production.     

Biogenic monoamines of the brain and spinal cord pia mater in vertebrates

Fluorimetric studies have been made on the content of adrenalin, noradrenaline, serotonin, dopamine, and tryptamine in the pial matter of the brain and spinal cord of fishes, birds and mammals including man. Using histochemical method with glyoxylic acid, biogenic monoamines were revealed in the adrenergic nerve fibers and monoaminocytes. Their total content in the pial matter of the brain is approximately the same in all vertebrates, being significantly lower in man. Higher concentration of adrenergic axons and lower amount of monoaminocytes in human subjects reveal the key role of the nervous influences in regulation of hemodynamics of the brain.     

Computer-operated microspectrofluorimetry to identify formaldehyde-induced fluorophores of biogenic monoamines and precursor substances in models and tissue sections

Summary By means of a histochemical reaction using formaldehyde vapour (Falck and Owman 1965), biogenic monoamines and precursor substances, i.e., L-DOPA, dopamine, noradrenaline, adrenaline, 5-hydroxytryptophan and 5-hydroxytryptamine, may be converted into fluorophores with specific spectral characteristics, i.e., the emission spectrum, excitation spectrum and fading curve. The registration and correction of the spectral properties and changes induced by acidification with hydrochloric acid vapour or treatment with ammonia vapour, of these formaldehyde-induced fluorophores, are performed by an automated microspectrofluorimeter, developed by modification of a Leitz MPV 2 system. This work deals with the instrumental configuration and certain methodological features in order to identify the fluorogenic biogenic monoamines and precursor substances in models and tissue sections. Registrations of excitation peak values, for the first time extended to a wavelength range from 240–460 nm, are discussed, which enable the calculation of peak ratio values 410/260, 380/320, 320/260 or 385/315, suitable as identification parameters for formaldehyde-induced fluorophores of biogenic monoamines and precursor amino acids.Dedicated to Mrs. Prof. Dr. M.H.A. De Groodt-Lasseel, founder of the Institute of Histology and Microscopic Anatomy of the University of Antwerp     

Monoamines in the nervous system of the tube-wormChaetopterus variopedatus (Polychaeta): Biochemical detection and serotonin immunoreactivity

Summary The presence and distribution of biogenic monoamines in the tube-wormChaetopterus variopedatus were investigated by a radioenzymatic method and HPLC with electrochemical detection, and the cellular localization of serotonin by peroxidase-antiperoxidase (PAP) immunohistochemistry with an antibody against serotonin-formaldehyde-protein conjugate. Dopamine, norepinephrine, epinephrine, serotonin (5-HT) and some of their metabolites were detectable, dopamine and norepinephrine being present in substantially larger amounts than 5-HT and epinephrine. With few exceptions, the largest amounts of amines were localized in the most nerve-rich tissues such as tentacles, and those containing cerebral ganglia and the ventral nerve cord. Serotonin-immunoreactive unipolar neurons were widely distributed in the dorso-lateral cerebral ganglia, the neurosecretory pharyngeal ganglion and the segmental ganglia of the anterior (dorsolateral) and posterior (medioventral) nerve cords. Some nerve-fiber tracts stained in the cerebral ganglia, but the neuropiles of segmental ganglia were the most intensely reactive CNS structures. Numerous reactive fibers were also present in connectives, commissures and segmental nerves. All peripheral sensory structures included serotonin-immunoreactive cells and neurites, especially the parapodial cirri and the bristle receptors of the setae. Trunk and parapodial muscles contained reactive varicose fibers and neuronal somata. These results suggest that monoamines are abundant and widespread in these worms and that 5-HT appears to have a key sensory role.     

Monoamine oxidase in adult Ascaridia galli

Monoamine oxidase (MAO), catalysing oxidative deamination of biogenic monoamines, has been detected in adult Ascaridia galli. MAO was present in mitochondria and deaminated noradrenaline at the maximal rate, although serotonin, adrenaline, tyramine and dopamine were also degraded but more slowly. Of the organs studied, the body wall, female reproductive organ and intestine, the body wall (containing neuronal structures) showed highest MAO activity. Km value for chick ascarid mitochondrial MAO using tyramine as substrate was 1.66 X 10(-3) M and it was most active at 2.5 mM tyramine concentration, pH 7.5 and 40 degrees C. MAO of A. galli appeared to be thermolabile as nearly 80% of its activity was lost when the incubation temperature was increased 5 degrees above optimum.     

Carotenoids in petals of perennial Medicago species

The carotenoid content in the petals of fourteen Medicago species was examined, together with eight species studied previously, caratenoids in all the known perennials of the genus are reported. The species can be arranged in relationship groups on the basis of their interfertility. No major carotenoid was species- or groupspecific; a few minor pigments, however, were group- or species-specific. The amount of carotenoids ranged from 7 μg/g dry matter in violet-flowered M. sativa to 2120 μg/g in brownish-yellow M. platycarpos. Xanthophylls constituted 76–99% of the total, with lutein as the major component. β-Carotene, lutein and flavoxanthin were ubiquitous in petals. In M sativa leaves β-carotene, lutein, violaxanthin and neoxanthin constituted 88% of the total. The xanthophylls were esterified in petals but not in leaves.     

Localization of biogenic monoamines in the dura mater of the rat brain

Quantitative and qualitative composition of catecholamines (noradrenaline and dophamine) and indolalkylamines (serotonin and tryptamine) and their localization have been studied in cells and neural fibers of the rat dura mater. Luminiscent-cytophotometric and electron microscopic methods have revealed two types of cells depositing biogenic monoamines. As demonstrate the experiments with rausedil injection, monoaminocytes of the first type (mast cells) contain predominantly indolalkylamines and a small amount of dophamine. Monoaminocytes of the second type (chromaffin cells) synthesize mainly serotonin and, perhaps, tryptamine. In adrenergic neural fibers only noradrenaline has been found to be present.     

Evidence for the existence of serotonin-, dopamine-, and noradrenaline-containing neurons in the gut of Lampetra fluviatilis

Summary Monoamine-containing neurons in the gut of Lampetra fluviatilis are characterized by histochemical, electron microscopical and biochemical methods. Strongly yellow fluorescent, probably serotonin-containing intrinsic neurons are found along the entire length of the intestine. Their processes aggregate to form large bundles of mainly non-terminal axons, constituting a subepithelial fibre plexus. This subepithelial, ganglion cell comprising plexus is connected to a wide-meshed subserosal plexus which has ganglion cells of different size and few varicose, single axons. Intermingled with both plexus there occur — in the anterior and middle but not in the preanal portion of the lamprey intestine — scattered green fluorescent intrinsic perikarya, emanating faintly green fluorescent, poorly varicosed axons.The formaldehyde-induced neuronal fluorophores conform to serotonin (yellow fluorescent compound), noradrenaline, and dopamine (green fluorescent substance), as revealed in microspectrofluorimetric recordings. The electron microscopical analysis of the yellow fluorescent intrinsic neurons in the terminal hindgut shows nerve cell pericarya and axons equipped with a typical population of occasional small granular and many large granular vesicles (750–1600 Å). The number and opacity of cores of the small and the osmiophilia of the cores of the large granular vesicles are significantly increased following short-term treatment with 5,6-dihydroxytryptamine. Long-term treatment with 5,6- or 5,7-dihydroxytryptamine provokes severe signs of ultrastructure impairment and eventual degeneration in the supposed serotonin-containing axons, besides indications of piling-up of organelles in the non-terminal axons due to arrest of axonal transport.Chromatography of acid extracts from the lamprey intestine, gills and kidney reveals the presence of serotonin (besides another unidentified indoleamine) and dopamine and noradrenaline in the gut, but only dopamine in the brain. The detection of serotonin, noradrenaline and dopamine in the lamprey gut is confirmed by chemical determinations.The occurrence of intrinsic serotonin-, noradrenaline- and dopamine-containing neurons in the gut of Lampetra fluviatilis deviates from the established pattern of innervation of the vertebrate intestine and is considered to be a remnant of an autonomic innervation principle common in invertebrates.Supported by grants from the Deutsche Forschungsgemeinschaft.Supported by grants from the Swedish Medical Research Council (No. 14X-712 and 14X-56.The authors are indebted to Lilan Bengtsson, Gertrude Stridsberg, Eva Svensson and Rolf Frank for skilful technical assistance.