Dopamine-immunoreactive Cells Research Articles

Effects of moderate alcohol consumption on behavior and neural systems of Wistar rats.

Chronic alcohol consumption affects various neurotransmitters, especially those implicated in the transitioning to alcohol use disorders (particularly dopaminergic and CRFergic systems). Few studies have investigated moderate alcohol consumption and its harmful consequences. The objective of this work was to analyze behavioral and neurochemical (dopaminergic and CRFergic systems) alterations during chronic moderate alcohol consumption. Twelve male Wistar rats were submitted to an intermittent alcohol ingestion protocol (alcohol group) for four weeks. The control group consisted of six rats. Open Field and Elevated Plus Maze tests were used for analysis of motor and anxiety-like behaviors. Immunohistochemistry analysis was performed in dopaminergic and CRFergic systems. Animals exposed to alcohol consumed moderate doses, chronic and intermittently. Behavioral tests detected fewer fecal boli in the alcohol exposed group, and immunohistochemical analysis indicated fewer dopamine-immunoreactive cells in the ventral tegmental area, and more CRF-immunoreactive cells in the anterior cingulate cortex and dorsolateral septum in this group. Thus we concluded that Wistar rats that consumed moderate doses of alcohol voluntarily and chronically showed a discreet anxiolytic effect in behavior, and a hypodopaminergic and hyperCRFergic neurochemical condition, which together are strong inducers of alcohol consumption predisposing to the development of alcohol use disorder (AUD).

Vesicular monoamine transporter 2 (SLC18A2) regulates monoamine turnover and brain development in zebrafish.

We aimed at identifying potential roles of vesicular monoamine transporter 2, also known as Solute Carrier protein 18 A2 (SLC18A2) (hereafter, Vmat2), in brain monoamine regulation, their turnover, behaviour and brain development using a novel zebrafish model. A zebrafish strain lacking functional Vmat2 was generated with the CRISPR/Cas9 system. Larval behaviour and heart rate were monitored. Monoamines and their metabolites were analysed with high-pressure liquid chromatography. Amine synthesising and degrading enzymes, and genes essential for brain development, were analysed with quantitative PCR, in situ hybridisation and immunocytochemistry. The 5-bp deletion in exon 3 caused an early frameshift and was lethal within 2weeks post-fertilisation. Homozygous mutants (hereafter, mutants) displayed normal low locomotor activity during night-time but aberrant response to illumination changes. In mutants dopamine, noradrenaline, 5-hydroxytryptamine and histamine levels were reduced, whereas levels of dopamine and 5-hydroxytryptamine metabolites were increased, implying elevated monoamine turnover. Consistently, there were fewer histamine, 5-hydroxytryptamine and dopamine immunoreactive cells. Cellular dopamine immunostaining, in wild-type larvae more prominent in tyrosine hydroxylase 1 (Th1)-expressing than in Th2-expressing neurons, was absent in mutants. Despite reduced dopamine levels, mutants presented upregulated dopamine-synthesising enzymes. Further, in mutants the number of histidine decarboxylase-expressing neurons was increased, notch1a and pax2a were downregulated in brain proliferative zones. Lack of Vmat2 increases monoamine turnover and upregulates genes encoding amine-synthesising enzymes, including histidine decarboxylase. Notch1a and pax2a, genes implicated in stem cell development, are downregulated in mutants. The zebrafish vmat2 mutant strain may be a useful model to study how monoamine transport affects brain development and function, and for use in drug screening.

On the larva and the zooid of the pterobranch Rhabdopleura recondita Beli, Cameron and Piraino, 2018 (Hemichordata, Graptolithina)

Hemichordates (Enteropneusta and Pterobranchia) belong to a small deuterostome invertebrate group that may offer insights on the origin and evolution of the chordate nervous system. Among them, the colonial pterobranch Rhabdopleuridae are recognized as living representatives of Graptolithina, a taxon with a rich fossil record. New information is provided here on the substrate selection and the life cycle of Rhabdopleura recondita Beli, Cameron and Piraino, 2018, and for the first time, we describe the nervous system organization of the larva and the adult zooid, as well as the morphological, neuroanatomical and behavioural changes occurring throughout metamorphosis. Immunohistochemical analyses disclosed a centralized nervous system in the sessile adult zooid, characterized by different neuronal subsets with three distinct neurotransmitters, i.e. serotonin, dopamine and RFamide. The peripheral nervous system comprises GABA-, serotonin-, and dopamine-immunoreactive cells. These observations support and integrate previous neuroanatomical findings on the pterobranch zooid of Cephalodiscus gracilis. Indeed, this is the first evidence of dopamine, RFamide and GABA neurotransmitters in hemichordates pterobranchs. In contrast, the lecithotrophic larva is characterized by a diffuse basiepidermal plexus of GABAergic cells, coupled with a small group of serotonin-immunoreactive cells localized in the characteristic ventral depression. It is envisaged the use of R. recondita as a novel and easily accessible hemichordate model organism to shed light on the evolution of hemichordates and more generally on the origin of deuterostome developmental mechanisms.

Dopaminergic modulation of spinal neurons and synaptic potentials in the lamprey spinal cord.

It has been shown previously that dopamine-immunoreactive cells and processes are present in the lamprey spinal cord and that dopamine modulates the cycle period of fictive swimming. The present study was undertaken to further characterize the effects of dopamine on the cellular properties of lamprey spinal neurons and on inhibitory and excitatory postsynaptic potentials to determine how dopaminergic modulation may affect the central pattern generator for locomotion. Dopamine reduced the late afterhyperpolarization (late AHP) following the action potential of motoneurons, and in three types of sensory neurons: dorsal cells, edge cells, and giant interneurons. The late AHP was not reduced in lateral interneurons or CC interneurons, both of which are part of the central motor pattern generating neural network. The reduction of the late AHP in motoneurons, edge cells, and giant interneurons resulted in an increase in firing frequency in response to depolarizing current injection. In the six cell classes examined, no changes were observed in the resting membrane potential, input resistance, rheobase, spike amplitude, or spike duration after application of dopamine. The durations of action potentials broadened by application of tetraethylammonium in motoneurons and of calcium action potentials in dorsal cells and giant interneurons were decreased after bath application of 10 microM dopamine. The durations of tetrodotoxin-resistant, N-methyl-D-aspartate-induced membrane potential oscillations in lamprey spinal motoneurons were increased after bath application of 1-100 microM dopamine, due perhaps to reduced calcium entry and thus reduced Ca(2+)-dependent K+ current responsible for the repolarization of the membrane potential during each oscillation. Polysynaptic inhibitory postsynaptic potentials (IPSPs) elicited in lamprey spinal motoneurons by stimulation of the contralateral half of the spinal cord were reduced by bath application of 10 microM dopamine. Polysynaptic excitatory postsynaptic potentials were not reduced by dopamine. Monosynaptic IPSPs in motoneurons elicited by stimulation of single contralateral inhibitory CC interneurons and single ipsilateral axons were reduced by bath application of dopamine (10 microM). Monosynaptic IPSPs in CC interneurons elicited by stimulation of ipsilateral lateral interneurons, however, showed no change after application of dopamine. The lack of dopaminergic effect on the late AHP of the locomotor network neurons, lateral interneurons and CC interneurons, and the selective reduction of IPSPs from CC interneurons suggest that synaptic modulation may play an important role in dopaminergic modulation of cycle period during fictive swimming in the lamprey.

Demonstration of dopamine immunoreactivity in open and closed type endocrine cells of gerbil (Meriones unguiculatus) stomach.

Dopamine-immunoreactive cells were identified in the stomach of gerbils using a selective antibody against glutaraldehyde-conjugated dopamine. In the pyloric area dopamine-containing cells were exclusively localized in the basal part of the mucous membrane, whereas in the fundic area dopaminergic cells were seen throughout the basodistal extent of the mucous membrane. Dopamine-containing cells belong to the class of closed and open type endocrine cells and were found in the fundic area and in the pyloric area respectively. These findings are discussed along with the likely modulatory function of dopamine in peripheral tissues.

The distribution of neurones immunoreactive for ?-tyrosine hydroxylase, dopamine and serotonin in the ventral nerve cord of the cricket, Gryllus bimaculatus

The cellular localization of the biogenic amines dopamine and serotonin was investigated in the ventral nerve cord of the cricket, Gryllus bimaculatus, using antisera raised against dopamine, β-tyrosine hydroxylase and serotonin. Dopamine-(n<-70) and serotonin-immunoreactive (n<-120) neurones showed a segmental arrangement in the ventral nerve cord. Some neuromeres, however, did not contain dopamine-immunoreactive cell bodies. The small number of stained cells allowed complete identification of brain and thoracic cells, including intersegmentally projecting axons and terminal arborizations. Dopamine-like immunostaining was found primarily in plurisegmental interneurones with axons descending to the soma-ipsilateral hemispheres of the thoracic and abdominal ganglia. In contrast, serotonin-immunostaining occurred predominantly in interneurones projecting via soma-contralaterally ascending axons to the thorax and brain. In addition, serotonin-immunoreactivity was also present in efferent cells and afferent elements. Serotonin-immunoreactive, but no dopamine-immunoreactive, varicose fibres were observed on the surface of some peripheral nerves. Varicose endings of both dopamine-and serotonin-immunoreactive neurones occurred in each neuromere and showed overlapping neuropilar projections in dorsal and medial regions of the thoracic ganglia. Ventral associative neuropiles lacked dopamine-like immunostaining but were innervated by serotonin-immunoreactive elements. A colocalization of the two amines was not observed. The topographic representation of neurone types immunoreactive for serotonin and dopamine is discussed with respect to possible modulatory functions of these biogenic amines in the central nervous system of the cricket.

Baroreceptor-aortic nerve-mediated release of endogenous L-3,4-dihydroxyphenylalanine and its tonic depressor function in the nucleus tractus solitarii of rats

We have proposed that l-3,4-dihydroxyphenylalanine ( l-DOPA) is a neurotransmitter and/or neuromodulator in the central nervous system [Misu Y. and Goshima Y. (1993) Trends pharmac. Sci. 14, 119–123]. This study aimed to explore whether or not endogenous l-DOPA, as a neurotransmitter candidate of the primary baroreceptor afferents, tonically functions to activate depressor neurons in the nucleus tractus solitarii of anesthetized rats. By parallel microdialysis in bilateral nucleus tractus solitarii areas, the basal l-DOPA release was in part inhibited by tetrodotoxin perfusion (1 μM) or Ca 2+ deprivation, and was markedly reduced by α-methyl- p-tyrosine(200mg/kg, i.p.), a tyrosine hydroxylase inhibitor. Forty to 100 mM K + concentration-dependently released l-DOPA. Fifty millimoles K + repetitively and constantly released l-DOPA. This release was Ca 2+-dependent. Stimulation of the left aortic nerve (100 Hz, 8 V) repetitively and constantly released l-DOPA and this release was tetrodotoxinsensitive. Phenylephrine i.v. infused produced l-DOPA release and reflex bradycardia, temporally associated with a rise and subsequent recovery of blood pressure. This release and bradycardia were abolished by denervation of the bilateral carotid sinus and aortic nerves. In addition, l-DOPA methyl ester, a competitive l-DOPA antagonist, when microinjected into depressor sites of the left nucleus tractus solitarii, antagonized depressor responses to mild stimulation (20 Hz, 3 V) of the ipsilateral aortic nerve. This antagonist alone, microinjected bilaterally, elicited a dose-dependent hypertension, which was abolished by α-methyl- p-tyrosine. Furthermore, by immunocytochemical analysis seven days after denervation of the left aortic nerve, tyrosine hydroxylase- and l-DOPA-, but not dopamine- and dopamine-β-hydroxylase-immunoreactivities decreased in the ipsilateral nucleus tractus solitarii and dorsal motor vagus nucleus complex area. In the left ganglion nodosum, denervation decreased staining and number of l-DOPA-immunoreactive cells and staining of tyrosine hydroxylase-immunoreactive cells, but no modification of dopamine-immunoreactive cells was seen. Taken together with previous findings that l-DOPA itself is stereoselectively responsible for cardiovascular control in this nucleus, it is probable that l-DOPA is a neurotransmitter of the primary baroreceptor afferents terminating directly in depressor neurons and/or indirectly in some neurons within a microcircuit, including depressor neurons of the nucleus tractus solitarii. Endogenously released l-DOPA itself tonically functions to activate depressor neurons for regulation of blood pressure in the rat nucleus tractus solitarii.

Distribution of tyrosine hydroxylase and dopamine immunoreactivities in the brain of the South African clawed frog Xenopus laevis.

The distribution of dopamine (DA) and the biosynthetic enzyme tyrosine hydroxylase (TH) has been studied immunohistochemically in the brain of the adult South African clawed frog, Xenopus laevis. The goals of the present study are, firstly, to provide detailed information on the DA system of the brain of a species which is commonly used in laboratories as an experimental model and, secondly, to enhance our insight into primitive and derived characters of this catecholaminergic system in amphibians. Dopamine-immunoreactive cell bodies are present in the olfactory bulb, the preoptic area, the suprachiasmatic nucleus, the nucleus of the periventricular organ and its accompanying cells, the nucleus of the posterior tubercle, the posterior thalamic nucleus, the midbrain tegmentum, around the solitary tract, in the ependymal layer along the midline of the caudal rhombencephalon, and along the central canal of the spinal cord. In contrast to the DA antiserum, the TH antiserum fails to stain the liquor-contacting cells in the periventricular organ. On the contrary, the latter antiserum reveals additional immunoreactive cell bodies in the olfactory bulb, the isthmic region and the caudal brainstem. Both antisera yield an almost identical distribution of fibers. Distinct fiber plexuses are observed in the olfactory bulb, the basal forebrain, the hypothalamus and the intermediate lobe of the hypophysis. Features that Xenopus shares with other anurans are the larger number of DAi cells, which are generally smaller in size than those observed in urodeles, and the lack of DAi fibers in pallial structures. On the other hand, the paired midbrain DA cell group and the innervation of the tectum of Xenopus resemble those found in the newt rather than those in frogs. Despite the existence of these species differences, the brain of Xenopus offers an excellent model for studying general aspects of neurotransmitter interactions and the development of catecholamine systems in this class of vertebrates.

Demonstration of dopamine-immunoreactive cells in the proximal convoluted tubule of gerbil (Meriones unguiculatus) kidney.

We investigated dopamine immunoreactivity in the kidney of gerbils (Meriones unguiculatus). For that purpose a sensitive and selective antibody against glutaraldehyde-conjugated dopamine was applied. Dopamine-immunoreactive cells were found in the epithelium of the proximal convoluted tubule, where these cells revealed a typical segment-like distribution pattern. Dopamine-immunoreactive precipitates were small and concentrated at the apical pole of the labeled cells. This study has directly identified dopamine as a constituent of certain cells of the proximal convoluted tubule in gerbils. The functional significance of dopamine in these cells is discussed in relation to the present view of renal dopaminergic actions.

Demonstration of dopamine-immunoreactive cells in the gastrointestinal tract of gerbils (Meriones unguiculatus).

We investigated dopamine immunoreactivity in the small intestine of gerbils (Meriones unguiculatus), using a sensitive and selective antibody against glutaraldehyde-conjugated dopamine. Dopamine-immunoreactive cells were found within the mucosal epithelium along the entire extent of the small intestine. Labeled cells were regularly distributed in the upper half of the intestinal villi, basally attached to the basement membrane and always reaching as far as the epithelial surface. Dopamine-containing cells revealed a spindle-like shape, and both light and electron microscopic characteristics relate them to typical open-type gut endocrine cells. Thus, this current study directly identified dopamine as a probable secretory product in basal granulated cells of the small intestine. The functional significance of these cells is discussed in relation to the current view of dopaminergic actions in peripheral tissues.

Distribution of serotonin- and dopamine-immunoreactivity in the brain of the teleost Clarias gariepinus

The distribution of serotonergic and dopaminergic cell bodies and varicose fibres in the brain of the teleost Clarias gariepinus was studied immunohistochemically using antisera against formaldehyde-conjugated serotonin and dopamine. Many serotonergic and dopaminergic fibres innervated the areas dorsalis telencephali pars medialis and pars lateralis dorsalis, as well as the area ventralis telencephali pars ventralis. In the diencephalon, a large number of serotonergic and some dopaminergic fibres were found in the preoptic nucleus, innervating the cells of this nucleus. In addition, serotonergic and dopaminergic fibres were observed in the pituitary stalk and in all regions of the pituitary gland. Moreover, the diencephalon contained the highest number of serotonin- or dopamine-immunoreactive cell bodies. These cells were confined to the same periventricular nuclei as the nucleus ventromedialis thalami, the nucleus posterior periventricularis, the nucleus lateralis tuberis, the nuclei recessus lateralis and recessus posterioris. Most cells of these nuclei were in contact with the cerebrospinal fluid of the third ventricle. The brainstem contained serotonergic cell bodies in the raphe nuclei and a few serotonergic and dopaminergic fibres. The torus semicircularis was densely innervated by serotonergic fibres and, to a lesser extent, dopaminergic fibres. In the midbrain of Clarias gariepinus, no dopaminergic homologue of the substantia nigra was observed. The results are discussed both in a comparative and a physiological context. In this regard, special attention has been paid to the contribution of hypothalamic monoamines in the regulation of gonadotropin secretion as an essential step in the neuro-endocrine control of reproduction.

Presence of dopamine-immunoreactive cell bodies in the catecholaminergic group A15 of the sheep brain

Antisera were raised in rabbits against dopamine or noradrenaline conjugated to thyroglobulin with glutaraldehyde. These antisera, tested in enzyme linked immunosorbent assay and immunohistochemistry specifically recognized their homologous antigens. With the aid of anti-tyrosine hydroxylase, anti-aromatic aminoacid decarboxylase, anti-dopamine-beta-hydroxylase, anti-dopamine, and anti-noradrenaline antisera, immunohistochemical reactions were performed on glutaraldehyde fixed sections of sheep diencephalon in order to determine the presence of dopamine in the catecholaminergic group A15. Perikarya of this nucleus were stained with anti-tyrosine hydroxylase, anti-aromatic aminoacid decarboxylase and anti-dopamine, but not with anti-dopamine-beta-hydroxylase or anti-noradrenaline. Both of these latter antisera stained fibers within this area. So as recently found in the rat, we could conclude that dopamine is present in group A15 of the sheep.

Host afferents into intrastriatal transplants of fetal ventral mesencephalon

Host afferents into fetal ventral mesencephalic tissue grafted to the neostriatum of adult rats have been studied by using anterograde transport of Phaseolus vulgaris leucoagglutinin (PHA-L) and immunocytochemistry for serotonin (5-HT), Substance P (SP), and dopamine-adenosine 3′:5′-monophosphate-regulated-phosphoprotein-32 (DARPP-32). Numerous fibers of cortical origin were detected in the transplants following multiple (11–15) iontophoretic injections of PHA-L into the frontal and anterior cingulate cortex. The labeled fibers occurred with an apparently random distribution throughout the graft tissue. Their overall density was lower than that of the surrounding striatum but similar to that found in the host nigra-ventral tegmental area. The majority of the PHA-L-labeled fibers in the grafts were thin and tortuous with varicosities or lateral clubs with terminal boutons. Dual labeling showed frequent close appositions between PHA-L-labeled terminals and dopamine-immunoreactive cell bodies. In parallel electron microscopy, synaptic contacts were observed between PHA-L-labeled terminals and unlabeled neuronal profiles in the graft. Other labeled fibers in the grafts were thick and smooth, corresponding probably to labeled myelinated axons observed in the electron microscope. These thick fibers were often seen to give off collaterals of the thin type. The virtual absence of such thick fibers in the normal striatal neuropil suggests that at least some of the cortical afferents to the grafts may have sprouted from axons normally projecting to diencephalic or brian stem regions. Serotonin fibers occurred in patches or as scattered single fibers in both deep and superficial portions of the nigral transplants. In the electron microscope some of these terminals were seen to establish synaptic contacts with nonimmunoreactive elements inthe graft. These fibers were present also when the graft tissue had been pretreated with 5,7-dihydroxytryptamine at the time of transplantation. This treatment eliminated all 5-HT-containing neurons from the grafts without any noticeable adverse effect on the survival of the dopaminergic neurons. The serotonin fibers in the grafts were thus most likely of host origin. SP-positive fibers formed a dense plexus inside the grafts. Since many SP-positive cell bodies were visualized inside the transplant after colchicine pretreatment, it is unclear, however, whether any of these fibers were of host origin. Intrastriatal injections of PHA-L or DARPP-32 immunocytochemistry indicated that the deep portions of the nigral grafts were entirely devoid of host striatal afferents. However, dual tyrosine hydroxylase and DARPP-32 immunocytochemistry showed close appositions between host striatal fibers and grafted dopamine neurons at the junction between the graft and the surrounding striatal tissue, suggesting that more extensive graft-host interaction may occur at the graft-host interface. These observations provide evidence that some of the projection systems available at the graft site can establish afferent connections with the grafted neurons, and they suggest that host cortical and brain stem afferents, in particular, may play a role in the functional regulation of the grafted dopaminergic neurons.

Distribution of dopamine immunoreactivity in the brain of the mormyrid teleost Gnathonemus petersii.

The distribution of dopamine-containing cell bodies and fibers was studied with aid of specific antibodies against dopamine in the highly developed brain of the weakly electric fish Gnathonemus petersii. In the telencephalon, dopamine-containing cell bodies were observed in a small area, i.e., area ventralis pars dorsalis and supracommissuralis. In the diencephalon, moderate numbers of dispersed dopamine-immunoreactive cells were present in the preoptic region, while large numbers of dopamine-containing neurons occurred in the hypothalamic paraventricular organ and neighbouring regions. The paraventricular organ, located around small (anterior, intermediate, and posterior) recesses contained many dopamine-immunoreactive cerebrospinal fluid-(CSF)-contacting neurons. Dopamine-containing cells were also observed in a magnocellular hypothalamic cell group, in the nucleus of the lateral recess, and in the nucleus posterior tuberis. In the mesencephalon only a few dopamine-containing cells were observed in a dorsal tegmental (possibly pretectal) area, whereas in ventral mesencephalic regions dopamine-containing cells were lacking. More caudally, dopamine-containing cells were observed in the presumed locus coeruleus, in the caudal region of the reticular formation, and in the presumed area postrema. Dopamine-immunoreactive fiber density was very high in the medioventral hypothalamus and in the preoptic region, where a dense subependymal plexus was observed along the preoptic recess. Such a plexus was also present in the caudal rhombencephalon, where it probably arises from the area postrema. Moderate numbers of dopamine-immunoreactive fibers were present in medioventral parts of the brain along its total rostrocaudal extent as well as in several subnuclei of the torus semicircularis, in the tectum mesencephali, and in the medial part of the dorsal telencephalic area. Other parts of the dorsal telencephalic area, as well as the large cerebellum and the electrosensory lateral line lobe of Gnathonemus, did not contain detectable amounts of dopamine. In spite of the high differentiation of the brain of Gnathonemus, the distribution of catecholamines as visualized with dopamine immunohistochemistry appears to be basically similar to that described in other teleostean and actinopterygian fishes on the basis of formaldehyde-induced fluorescence or tyrosine hydroxylase immunohistochemistry.(ABSTRACT TRUNCATED AT 400 WORDS)

Immunohistological evidence of dopamine cells in the cephalic nervous system of the silkworm Bombyx mori. Coexistence of dopamine and ? endorphin-like substance in neurosecretory cells of the suboesophageal ganglion

Evidence of dopamine cells in the brain and the suboesophageal ganglion of the silkworm Bombyx mori was obtained immunohistologically in larvae and pupae. From six to eight and eight (two symmetrical groups of four) immunoreactive cells are present respectively in median and lateral protocerebral areas of the brain. In the suboesophageal ganglion, two cell clusters with dopamine immunoreactivity were observed. There was no clear difference in the nature of the immunohistochemical reaction and the number of cells between diapause- and non-diapause-egg producers, in both brains and suboesophageal ganglia. By examination of adjacent sections, it was possible to show that dopamine-immunoreactive cells in larval suboesophageal ganglia also contain an α endorphin-like substance.