Dopamine as trace amine in the dorsal root ganglia

It has been shown that in the chick dorsal root ganglion (DRG) about 8% of neurons, belonging to both the A and B classes of sensory neurons exhibit a clear dopamine immunoreactivity. In the present study are reported the results of measurements, by mean of HPLC-electrochemical detection (HPLC-ED), of DA and of the DA metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the rat DRG and their central nerves. Very low levels of DA, about 10 folds lower than the levels found in the dorsal horn of the spinal cord, were found in the DRG. However the levels of DOPAC and HVA were approximately equivalent to the levels found in the cord. The immunocytochemical study performed in parallel has shown that some dopaminergic-immunoreactive fibers in the DRG are located around the blood vessels. Few dopamine-immunoreactive sensory neurons were identified in the DRG and immunoreactive fibers, not linked to blood vessels, were identified in the dorsal root nerves. The present work indicates that there is a dopaminergic innervation of the blood vessels in the rat DRG but that dopamine may also be, as in the chick, a transmitter of primary afferent fibers.     

Good riddance to dopamine: Roles for the dopamine transporter in synaptic function and dopamine-associated brain disorders

The neurotransmitter dopamine (DA) plays a critical role in CNS circuits that provide for attention, executive function, reward responses, motivation and movement. DA is inactivated by the cocaine- and amphetamine-sensitive DA transporter (DAT), a protein that also provides a pathway for non-vesicular DA release. After a brief review of DAT function and psychostimulant actions, we consider the importance DAT in relation to the distinct firing patterns of DA neurons that permit awareness of novelty and reward. Finally, we review recent efforts to gather direct support for DAT-linked disorders, with a specific focus on DAT mutations recently identified in subjects with ADHD.     

A quantitative analysis of the biogenic amines in the central ganglia of the pond snail,Lymnaea stagnalis (L.)

Biogenic amines in the central ganglia of Lymnaea stagnalis have been identified, quantified and localised using the techniques of high performance liquid chromatography (HPLC), radioenzymatic assay (REA) and fluorescence microscopy. HPLC indicated the presence of dopamine (DA), (5-hydroxytryptamine (5-HT), noradrenaline (NA) and 3,4-dihydroxyphenylacetic acid (DOPAC) in Lymnaea central nervous tissue. REA confirmed the presence of DA and NA in this tissue and, in addition, indicated that some adrenaline (Adr) may be present. Quantitatively, DA and 5-HT were the most significant amines detected, with the pedal ganglia containing the highest concentrations of both. High degrees of variation in DA and 5-HT concentrations were observed, both between animals from within the same sample group and between the mean values determined from separate groups. Whilst there was no obvious explanation for the differences between the sample group means, some evidence accrued to suggest that underlying seasonal variability may have been a contributory factor.     

An analysis of the inhibitory responses of dopamine and octopamine on Helix central neurons

1. Electrophysiological recordings were made from identified neurons in the isolated suboesophageal ganglionic mass of Helix aspersa. Cells were voltage clamped at testing membrane potential. 2. Bath addition of 1 microM dibutyryl cAMP caused a time dependent enhancement of an evoked IPSP and the dopamine (DA) and octopamine (OA) induced outward currents obtained in these neurons. Forskolin, 0.1 microM, which enhances and MDL 12,330A, 0.12 microM, which depresses adenylate cyclase activity also modified these responses. 3. The DA and OA inhibitory responses were both shown to be potassium mediated events. They were preferentially antagonised by low micromolar concentrations of 4-aminopyridine. Two other potassium channel antagonists, tetraethylammonium and apamin had little effect on the DA and OA responses. 4. Cell sensitivity to DA and OA was greatly enhanced in calcium free/2 mM cobalt Ringer. The reversal potential of the DA response was shifted to a more negative value in calcium free Ringer. Sodium free Ringer was also found to alter the responses to DA or OA but those results were not consistent.     

Geographic distribution of suitable hosts explains the evolution of specialized gentes in the European cuckoo Cuculus canorus

Background

We recently discovered two composite long terminal repeat (LTR)-retrotransposon-like elements which we named DA (~300 kb) and Xiao (~30 kb), meaning big and small in Chinese respectively. Xiao and DA (three types of DA identified) were found to have been derived from several donor sites and have spread to 30 loci in the human genome, totaling to 5 Mb. Our bioinformatics analyses with the released human, chimp, rhesus macaque, orangutan, and marmoset genomic sequences indicate that DA and Xiao emerged ~25 million years (Myr) ago.

Results

To better understand the evolution of these two complex elements, we investigated various internal junctions of DA and Xiao as well as orthologous genomic sites of the 30 DA/Xiao loci in non-human primates including great apes, lesser apes, Old World monkeys, New World monkeys, and a prosimian. We found that Xiao and type I DA first emerged in the genome between 25 and 18 Myr ago, whereas type II and Type III DAs emerged between 14 and 7 Myr ago. Xiao and DA were most active in great apes, with their amplification peaking during 25-14 and 14-7 Myr ago, respectively. Neither DA nor Xiao seem to have been active in the human and chimp genomes during last 6 Myr.

Conclusion

The study has led to a more accurate age determination of the DA and Xiao elements than our previous bioinformatics analyses, and indicates that the amplification activity of the elements coincided with that of group I HERV-Es during evolution. It has also illustrated an evolutionary path with stepwise structural changes for the elements during past 25 Myr, and in doing so has shed more light on these two intriguing and complex elements that have reshaped our genome.     

Characterization of seven DA/DAPI-positive bisatellited marker chromosomes by in situ hybridization

Summary Seven dicentric bisatellited marker chromosomes, ascertained at amniocentesis, chorionic villus sampling, and in blood from an abnormal liveborn were characterized cytogenetically. All seven markers demonstrated brilliant bands by the DA/DAPI technique corresponding to C-band positive regions. Although some dicentric DA/DAPI-positive bisatellited markers have been identified as inverted duplicated 15s, recent literature has suggested that DA/DAPI lacks specificity for chromosome 15. Our evaluation of DA/DAPI-positive bisatellited marker chromosomes by in situ hybridization shows that some originate from chromosome 15 whereas DA/DAPI negative bisatellited markers may not be derived from 15. The morphological variations noted in our studies are discussed with respect to nomenclature.     

A10 dopamine neurons: role of autoreceptors in determining firing rate and sensitivity to dopamine agonists

The present experiments investigated the relationship between the spontaneous basal firing rate of A10 dopamine (DA) neurons and their sensitivity to the rate-suppressant effects of intravenously administered apomorphine (APO) and d-amphetamine (AMP) as well as microiontophoretically ejected DA. The results indicated highly significant inverse relationships between basal neuronal activity and sensitivity to DA and DA agonists, i.e. the faster the spontaneous rate of an A10 DA neuron, the less sensitive that cell was to agonist-induced suppression. This relationship was not found for the rate suppressant effects of iontophoretic gamma-aminobutyric acid. There were no significant differences between the effects of iontophoretic DA on pre-glutamate and glutamate-driven activity of the same A10 DA neurons indicating that faster firing rates, per se, did not determine the sensitivity of these cells to DA agonists. Rather, these results suggest that both spontaneous activity and sensitivity to DA agonists may be determined by the density (or sensitivity) of DA autoreceptors on A10 DA neurons. This hypothesis was supported by the finding that antidromically identified mesocortical DA neurons, which were significantly less responsive to DA, APO and AMP exhibited significantly faster firing rates than other A10 DA neurons. Thus, this subpopulation of A10 DA neurons is primarily made up of cells with low autoreceptor density (or sensitivity).     

Pseudo-nitzschia in New Zealand and the role of DNA probes and immunoassays in refining marine biotoxin monitoring programmes

Domoic acid (DA) was first detected in shellfish in New Zealand after the implementation of a comprehensive biotoxin monitoring programme for amnesic, paralytic, diarrhetic and neurotoxic shellfish toxins, following a suspected neurotoxic shellfish poisoning (NSP) event in early 1993. Both phytoplankton monitoring and shellfish flesh testing programmes have led to an extensive database which has helped link species of Pseudo-nitzschia to specific DA outbreaks. In 1994, P. pungens and P. turgidula were associated with DA contamination of shellfish, and cultured isolates of these species proved to be toxin producers. During 1996 the use of species-specific ribosomal RNA (rRNA)-targeted oligonucleotide probes and DA immunoassays led to the discovery of toxin production by P. fraudulenta, and showed the nontoxic P. heimii to be a major bloom former. Pseudo-nitzschia delicatissima, P. pseudodelicatissima and P. multiseries, also identified using rRNA-targeted probes, have been linked to DA contamination of New Zealand shellfish; P. australis is the main cause of DA in scallops. The relative amnesic shellfish poisoning (ASP) risk associated with different species, largely determined by DA immunoassays of cultured isolates, is now used by some regulators to refine risk assessments. Species identification is therefore vital so that shellfish growers, and health and industry officials, can make safe and economically sound harvesting decisions. The development and field trialling of DNA probes is proving invaluable in this context.     

Multiple receptors for brain dopamine in behavior regulation: concept of dopamine-E and dopamine-I receptors

From current knowledge, it is possible to substantiate the original concept of DAe and DAi receptors including the predicted correlation with the anatomical, histochemical, biochemical and functional features of the distinct neuronal structures, in which they occur; labelling them as neostriatal DAe receptors and mesolimbic DAi receptors appears to be justified. Available data warrant a revision of currently employed behavior and pharmacological tests. When revised in terms of the DAe-DAi concept, assessment of such tests reveals that agents such as (-)NPA, 6, 7ADTN and certain ergot alkaloids like lergotrile, lisuride and, to a less degree, CB-154 are weak DAe agonists and strong DAi antagonists (Table I). The discovery that mesolimbic α-like NE receptors which regulate the DA activity at the level of the mesolimbic DAi, but not neostriatal DAe, receptors show adaptational changes following priming or subacute treatments with apomorphine or haloperidol, opens new perspectives for understanding phenomena such as the development of hypersensitivity to apomorphine etc. Presynaptic DA receptors located within the DA synaps and DA receptors located at DA cell-bodies resemble closely the DAe receptors, although absence of linkage to the enzyme adenylyl cyclase hints at some distinction. It is possible that the distinct classes of DA receptors identified by behavior and pharmacological studies in mice correspond with the DAe and DAi receptors in snails, rats and cats. There is no evidence to suggest that DAe and DAi receptors are directly related to a) so-called DA1 and DA2 receptors which are coupled and uncoupled respectively to the enzyme adenylyl cyclase, or to b) any of the DA-specific binding sites identified with radiolabelled DA agonists or antagonists. Nonetheless, it cannot be excluded that DAe receptors may correspond with DA-specific binding sites identified with tritiated DA and/or haloperidol, and DAi receptors with a particular subclass of DA-specific binding sites identified within certain mesolimbic structures with radiolabelled spiperone. Thus, future work is still required to relate DAe and DAi receptors to particular, molecular entities within the brain.     

Mutations in the zebrafish unmask shared regulatory pathways controlling the development of catecholaminergic neurons

The mechanism by which pluripotent progenitors give rise to distinct classes of mature neurons in vertebrates is not well understood. To address this issue we undertook a genetic screen for mutations which affect the commitment and differentiation of catecholaminergic (CA) [dopaminergic (DA), noradrenergic (NA), and adrenergic] neurons in the zebrafish, Danio rerio. The identified mutations constitute five complementation groups. motionless and foggy affect the number and differentiation state of hypothalamic DA, telencephalic DA, retinal DA, locus coeruleus (LC) NA, and sympathetic NA neurons. The too few mutation leads to a specific reduction in the number of hypothalamic DA neurons. no soul lacks arch-associated NA cells and has defects in pharyngeal arches, and soulless lacks both arch-associated and LC cell groups. Our analyses suggest that the genes defined by these mutations regulate different steps in the differentiation of multipotent CA progenitors. They further reveal an underlying universal mechanism for the control of CA cell fates, which involve combinatorial usage of regulatory genes.     

2株产纤维素酶放线菌的筛选及分类鉴定

从海南热带生境中采集样品14份,以CMC-Na为唯一碳源的培养基,筛选获得纤维素酶活性较高的放线菌DA08506和DA08602,CMC酶活力分别为276.642U/mL、773.982U/mL,FPA酶活力分别为82.638U/mL、325.674U/mL。通过多相分类,初步鉴定菌株DA08506可能是Streoptomyces的一个新种;菌株DA08602鉴定为Streptomyces chartreusis。     

Genome-wide linkage analysis of chronic relapsing experimental autoimmune encephalomyelitis in the rat identifies a major susceptibility locus on chromosome 9

The immunization of inbred Dark Agouti (DA) rats with an emulsion containing homogenized spinal cord and CFA induces chronic relapsing experimental autoimmune encephalomyelitis (EAE), a disease with many similarities to multiple sclerosis. We report here the first genome-wide search for quantitative trait loci regulating EAE in the rat using this model. We identified one quantitative trait locus on chromosome 9, Eae4, in a [DA(RT1av1) x BN(RT1n)]F2 intercross showing linkage to disease susceptibility and expression of mRNA for the proinflammatory cytokine IFN-gamma in the spinal cord. Eae4 had a larger influence on disease incidence among rats that were homozygous for the RT1av1 MHC haplotype (RT1av1 rats) compared with RT1n/av1 rats, suggesting an interaction between Eae4 and the MHC. Homozygosity for the DA allele at markers in Eae4 and in the MHC was sufficient for EAE. Thus, Eae4 is a major genetic factor determining susceptibility to EAE in this cross of DA rats. In addition, there was support for linkage to phenotypes of EAE on chromosomes 1, 2, 5, 7, 8, 12, and 15. The chromosome 12 region has been shown previously to predispose DA rats to arthritis, and the chromosome 2 region is syntenic to Eae3 in mice. We conclude that Eae4 and probably the other identified genome regions harbor genes regulating susceptibility to neuroinflammatory disease. The identification and functional characterization of these genes may disclose critical events in the pathogenesis of multiple sclerosis; understanding these events could be essential for the development of new therapies against the disease.     

Putative nociceptive neurotransmitters in the mesencephalic reticular formation

The neurotransmitter(s) involved in the transmission of nociceptive information in the mesencephalic reticular formation (MRF) of the rat have not been identified. Acetylcholine (ACh), substance P (SP), neurotensin (NT), norepinephrine (NE) and dopamine (DA) have all been implicated as putative neurotransmitters involved in nociception. All of these compounds were microiontophoretically administered in the MRF of rats to determine which, if any, mimicked the effects produced by a nociceptive stimulus (foot pinch). This is only one of several criteria that a substance should meet to be considered a nociceptive neurotransmitter in the MRF. ACh and NE mimicked the effects of the nociceptive stimulus in 61% and 67% respectively of the cells tested; NT, DA and SP mimicked the effects of the nociceptive stimulus less frequently (33%, 30%, 23% respectively). Therefore, the nociceptive neurotransmitters in the MRF appear to be ACh and NE; NT, DA and SP may be neurotransmitters with a less important role in nociception in the MRF.     

A genetic screen for behavioral mutations that perturb dopaminergic homeostasis in mice

Disruption of dopaminergic (DA) systems is thought to play a central role in the addictive process and in the pathophysiology of schizophrenia. Although inheritance plays an important role in the predisposition to these disorders, the genetic basis of this is not well understood. To provide additional insight, we have performed a modifier screen in mice designed to identify mutations that perturb DA homeostasis. With a genetic background sensitized by a mutation in the dopamine transporter (DAT), we used random chemical mutagenesis and screened for mutant mice with locomotor abnormalities. Four mutant lines were identified with quantitatively elevated levels of locomotor activity. Mapping of mutations in these lines identified two loci that alter activity only when dopamine levels are elevated by a DAT mutation and thus would only have been uncovered by this type of approach. One of these quantitative trait loci behaves as an enhancer of DA neurotransmission, whereas the other may act as a suppressor. In addition, we also identified three loci which are not dependent on the sensitized background but which also contribute to the overall locomotor phenotype.     

Projection-specific modulation of dopamine neuron synapses by aversive and rewarding stimuli

Midbrain dopamine (DA) neurons are not homogeneous but differ in their molecular properties and responses to external stimuli. We examined whether the modulation of excitatory synapses on DA neurons by rewarding or aversive stimuli depends on the brain area to which these DA neurons project. We identified DA neuron subpopulations in slices after injection of "Retrobeads" into single target areas of adult mice and found differences in basal synaptic properties. Administration of cocaine selectively modified excitatory synapses on DA cells projecting to nucleus accumbens (NAc) medial shell while an aversive stimulus selectively modified synapses on DA cells projecting to medial prefrontal cortex. In contrast, synapses on DA neurons projecting to NAc lateral shell were modified by both rewarding and aversive stimuli, which presumably reflects saliency. These results suggest that the mesocorticolimbic DA system may be comprised of three anatomically distinct circuits, each modified by distinct aspects of motivationally relevant stimuli.