Distribution Of Catecholamines Research Articles

Dopamine-beta-hydroxylase-like immunoreactive neurons in two insect species, Calliphora erythrocephala and Periplaneta americana.

The localization of dopamine-beta-hydroxylase in the cephalic central nervous system of the blowfly (Calliphora erythrocephala) and the cockroach (Periplaneta americana) was investigated. Immunoreactive neurons were demonstrated in both species. The results were compared with the known distribution of catecholamines in the brain of both species. In certain cell groups and neuropilar regions of both species D beta H-immunoreactivity coincides with the presence of catecholamines. Additionally D beta H immunoreactivity was found in several cell bodies and neuropilar regions in which no catecholamines could be detected. A correlation between the presence of octopamine and anti-D beta H labelling was not found. Thus it seems that the D beta H-immunoreactivity neither indicates the presence of octopamine nor is it limited to noradrenaline-containing neurons. Parallel findings in vertebrates are discussed.

Vicious Cycle of Catecholamines and K+ in Cardiac Ischemia

The distribution of catecholamine (CA)-containing neurones and tissue noradrenaline (NA) concentration was investigated in pig hearts with and without myocardial ischemia. All hearts were shock-frozen using a Wollenberger clamp, and tissue samples were obtained from the mid- and apical left ventricular wall (nonischemic and ischemic areas, respectively, in hearts with coronary artery occlusion). The distribution of CA-containing neurones was assessed morphometrically from histofluorescence sections, and the NA content measured using high pressure liquid chromatography (HPLC). In hearts without myocardial ischemia, both the density of CA-containing neurones (0.96 +/- 0.07 compared with 1.09 +/- 0.07% of total field area) and the NA content (738 +/- 63 compared with 884 +/- 84 ng/g wet weight, p less than 0.025) were less in apical samples compared with those obtained at the midventricular level. These differences were more pronounced in samples from hearts obtained following 5 min (fluorescence, 0.96 +/- 0.19 compared with 1.38 +/- 0.18%, p less than 0.01; NA content, 543 +/- 109 compared with 816 +/- 82 ng/g wet weight, p less than 0.01) or 15 min (fluorescence, 0.77 +/- 0.07 compared with 1.12 +/- 0.09%; NA content, 521 +/- 43 compared with 753 +/- 79 ng/g wet weight, p less than 0.01) of coronary artery occlusion, indicating a release of CA in the ischemic myocardium.

Distribution of catecholamines in the brain stem and spinal cord of the lizard Varanus exanthematicus: An immunohistochemical study based on the use of antibodies to tyrosine hydroxylase

Antibodies to tyrosine hydroxylase were used to study the distribution of nerve cells, fibers and terminals, containing catecholamines, in the lizard Varanus exanthematicus, by means of the indirect immunofluorescence technique. Tyrosine hydroxylase-containing cell bodies occurred in the hypothalamus, the ventral and dorsal tegmentum mesencephali, the substantia nigra, the isthmic reticular formation, in and ventrolaterally to the locus coeruleus, in the nucleus tractus solitarii and in a lateral part of the nucleus reticularis inferior. In addition tyrosine hydroxylase-containing cell bodies were found throughout the spinal cord, ventral to the central canal. Tyrosine hydroxylase-immunoreactive terminal areas in the brain stem were seen in the nucleus interstitialis of the fasciculus longitudinalis medialis, the nucleus raphes superior, the locus coeruleus, several parts of the reticular formation and the nucleus descendens nervi trigemini. Ascending catecholaminergic pathways could be traced from the ventral mesencephalic tegmentum as well as from the dorsal isthmic tegmentum rostralwards, through the lateral hypothalamus. These pathways correspond to the mesostriatal and isthmocortical projections respectively, as described in mammals. Furthermore, ascending catecholaminergic fibers could be traced from the catecholaminergic cell groups in the medulla oblongata to the isthmus, where they intermingle with the locus coeruleus neurons. These pathways correspond to the medullohypothalamic projection and to the dorsal periventricular system in mammals. Descending catecholaminergic fibers to the spinal cord pass via the dorsomedial part of the lateral funiculus, and mainly terminate in the dorsal horn. The results obtained in the present study have been placed in a comparative perspective, which illustrates the constancy of catecholaminergic innervation throughout phylogeny.

Immunohistochemical localization of dopamine in the brain of the insect Locusta migratoria migratorioides in comparison with the catecholamine distribution determined by the histofluorescence technique

As part of a follow-up study to our previous investigation of the catecholaminergic neurosecretory cells in the brain of adult female locusts (Locusta migratoria migratorioides) by means of the formaldehyde-induced fluorescence method, we have attempted to specify the identity of the amines present in these cells by an immunohistological technique. Using a recently developed anti-dopamine serum, we have demonstrated that the majority of the catecholaminergic median neurosecretory cells contain dopamine. Moreover, dopamine is present in some cell bodies of other zones of the brain, i.e. the median subocellar neurosecretory cells, perikarya in external areas of the protocerebrum, below the calyces, around the pedunculus, in the optic lobes (between the lobula and the medulla, between the medulla and the lamina), and in external zones of the tritocerebrum. Among the structured neuropils, which were particularly fluorescent in the formaldehyde-induced fluorescence method, only the pedunculus, the posterior part of the central body, the external zones of the alpha- and beta lobes and the proximal part of the lamina contain little dopamine.

Predominance of l-dopa in fetal plasma and the amniotic fluid during late gestation in the rat

The purpose of this study was to reevaluate catecholamine distribution in fetal and maternal compartments during late gestation in the rat. Fetal and maternal plasma and amniotic fluid were collected from anesthetized rats on consecutive days from day 17 to day 22, the day of parturition. The fluid was analyzed for dihydroxyphenylalanine (l-dopa), dopamine, norepinephrine, and epinephrine by radioenzymatic assays. Amniotic fluid volume was determined by a direct weighing method. l-Dopa concentrations constituted approximately 50% of total fetal plasma catecholamines and were significantly higher in fetal than in maternal circulation. Dopamine concentrations in fetal plasma were tenfold lower than those of l-dopa but were also significantly higher in fetal than in maternal plasma; norepinephrine levels were similar in both. Maternal plasma epinephrine levels remained relatively constant, whereas fetal epinephrine levels increased fiftyfold from day 17 to day 22. l-Dopa concentrations in the amniotic fluid were tenfold higher than those of dopamine, and the concentrations of both increased markedly during the last 2 days of gestation. However, this apparent rise could be attributed to the concomitant fivefold reduction in the amniotic fluid volume observed at this time. It is concluded that l-dopa is the predominant catecholamine in both the fetal plasma and the amniotic fluid during late gestation in the rat. At the present time, neither the source nor the possible physiologic functions of l-dopa during fetal life are known.

Regional distribution of free and sulfoconjugated catecholamines in the bovine adrenal cortex and medulla

The intraadrenal distribution of free and sulfoconjugated catecholamines and the activity of the catecholamine sulfoconjugating enzyme phenolsulfotransferase in the bovine adrenal gland are described. In the adrenal cortex all three free catecholamines, epinephrine, norepinephrine, and dopamine, were detected in various concentrations. In the adrenal medulla, the relative proportion of the main adrenal catecholamines, free epinephrine and free norepinephrine, was found to be higher in the medullary sections derived from the subcortical areas where the ratio of epinephrine to norepinephrine was 8:1 than in the centre of the organ where norepinephrine levels reached those of epinephrine. Free dopamine, representing about 1.0% of the total catecholamine content was distributed in the same pattern as epinephrine or the sum of epinephrine and norepinephrine, revealing a significant positive correlation between regional dopamine and epinephrine plus norepinephrine (r = 0.97) as well as between dopamine and epinephrine (r = 0.96). Phenolsulfotransferase activity was present in both medulla and cortex. Dopamine sulfate was detected in relatively small concentration in all cortical and medullary layers, but norepinephrine sulfate and epinephrine sulfate were not present. The meaning of this distribution of individual catecholamines and phenolsulfotransferase activity in both cortex and medulla is discussed.

Adrenal medullary implants in the dopamine-denervated rat striatum. I. Acute catecholamine levels in grafts and host caudate as determined by HPLC-electrochemistry and fluorescence histochemical image analysis

Rats with unilateral 6-hydroxydopamine-induced degeneration of the left nigrostriatal dopamine system were given intrastriatal implants of one cortex-free adrenal medulla divided into 4 pieces. Two pieces were placed in the center of the anterior part of the denervated caudate and two pieces in a more posterior position in lateral caudate. The distribution of catecholamines (CA) in grafts and host brain was studied 2, 100 and 400 min after grafting by HPLC-electrochemistry and Falck-Hillarp fluorescence histochemistry combined with computer-aided image analysis. Two minutes after implantation the chromaffin tissue grafts contained large amounts of adrenaline (A) and noradrenaline (NA) and small amounts of dopamine (DA). The chromaffin cells had a relatively normal fluorescence histochemical appearance. From the grafts, CA had spread into the surrounding host brain tissue where high levels of A and NA and low levels of DA were now found in the denervated host striatum. Fluorescence histochemistry and image analysis showed the CA to have spread 1–1.5 mm in all directions from the grafts. The CA concentrations decreased almost linearly with increasing distance from the grafts. At 100 min after implantation approximately a third of the chromaffin cells were still strongly fluorescent while the rest of the cells were very weakly fluorescent or non-fluorescent. The amounts of A, NA and DA in the host brain had decreased considerably, while the size of the fluorescent halo around the grafts had not diminished. At 400 min after grafting, only scattered cells in the chromaffin implants were strongly fluorescent and the surrounding host striatum contained low amounts of CA. It is concluded that intrastriatal adrenal medullary implants acutely release or leak large amounts of CA into surrounding host brain tissue. Taken together with results from the accompanying paper 24 these data show that the grafts can maintain CA levels in host striatum high enough to elicit strong rotational responses during approximately 200 min.

Chromogranin A, the major catecholamine storage vesicle soluble protein. Multiple size forms, subcellular storage, and regional distribution in chromaffin and nervous tissue elucidated by radioimmunoassay.

Chromogranin A (CgA), the major catecholamine storage vesicle (CSV) soluble protein, may index exocytotic sympathoadrenal secretion. To explore CgA in adrenergic tissues, we developed a radioimmunoassay for bovine CgA. Within adrenal medulla CSV, several minor chromogranins had similar amino acid compositions and peptide maps to that of CgA and also showed parallel, partial cross-reactivity in the CgA radioimmunoassay. CgA immunoreactivity represented 7 +/- 1% of total adrenal medulla cell protein and was localized to adrenal CSV, representing 46 +/- 2% of CSV soluble protein. In brain, there was 1000-fold less CgA than in adrenal medulla, with a widespread regional distribution (maximal in neocortex) and an unusual subcellular distribution (maximal in cytosol), both of which differ from reported catecholamine distribution. Brain chromogranin immunoreactivity also had a lower Stokes radius than adrenal CgA. Sympathetic nerve and serum had 6,000-fold and 30,000-fold less CgA than that in adrenal medulla. The results suggest a "family" of adrenal medulla chromogranins, similar structurally and immunoligically. Adrenal medulla and brain chromogranin differ in concentration, subcellular localization, and molecular size. Finally, CgA in serum may provide a useful tool for sympathoadrenal studies in intact organisms.

Bombesin-, gastrin/CCK-, 5-hydroxytryptamine-, neurotensin-, somatostatin-, and VIP-like immunoreactivity and catecholamine fluorescence in the gut of the elasmobranch, Squalus acanthias.

The presence of peptides and 5-hydroxytryptamine (5-HT) in neurons and endocrine cells in the gastrointestinal tract of the spiny dogfish, Squalus acanthias, was investigated by means of immunohistochemistry, and the distribution of catecholamines by use of the Falck-Hillarp fluorescence-histochemical technique. Bombesin-like immunoreactivity was present in numerous nerves in all layers and all parts of the gut, and also in endocrine cells in the mucosa throughout the stomach, rectum and intestine. VIP-like immunoreactivity occurred in an abundance of nerve fibres and in nerve cell bodies in all parts of the gut except the oesophagus, while 5-HT-like immunoreactivity was found sparsely in nerve fibres and more frequently in endocrine cells throughout the gut. Gastrin/CCK-like immunoreactivity was present in numerous nerve fibres in the rectum, but only in scattered fibres in the other parts of the gut. Endocrine cells showing gastrin/CCK-like immunoreactivity were present in the intestine only. Somatostatin-like immunoreactivity occurred in both nerve fibres and endocrine cells of the stomach and intestine, but only in nerves in the rectum. Neurotensin-like immunoreactivity was confined to endocrine cells of the intestine. Falck-Hillarp fluorescence histochemistry revealed 5-HT in endocrine cells and catecholamines in nerve fibres (and possibly also in endocrine cells) throughout the gut. Bombesin-, VIP-, gastrin/CCK- and somatostatin-like immunoreactivities and catecholamine fluorescence were present in nerve fibres of the rectal gland and, with the exception of gastrin/CCK-like immunoreactivity, also in nerve bundles in the walls of the coeliac and mesenteric arteries. The findings of the present study form an anatomical basis for the assumption that several of the neuropeptides and amines could function as neurotransmitters or neuromodulators in the gut of Squalus.

Distribution of catecholamines in individual hypothalamic nuclei of Japanese quail.

The dissection of eight hypothalamic nuclei or areas of Japanese quail brain (nucleus hypothalamicus medialis anterior, nucleus periventricularis magnocellularis, nucleus hypothalamicus lateralis anterior, nucleus hypothalamicus medialis posterior, nucleus hypothalamicus lateralis posterior, eminentia medialis, nucleus tuberis and area hypothalamica posterior) and a subsequent radioenzymatic measurement of the basic concentrations of noradrenaline, adrenaline and dopamine in these structures of female Japanese quail were demonstrated in this study. Individual hypothalamic nuclei were identified according to the histological map prepared. All the above mentioned structures were isolated from serial 200 micrometers thick frozen coronal sections of the hypothalamus under a stereomicroscope by means of a needle punch of 500 micrometers or 300 micrometers internal diameter. The highest levels of all biogenic amines measured were found in the area hypothalamica posterior which significantly differed from all other nuclei examined. Their lowest concentrations occurred in the nucleus hypothalamicus medialis posterior and in the nucleus hypothalamicus lateralis posterior. The results present the first picture of the quantitative distribution of catecholamines in the hypothalamus of the female Japanese quail.

Catecholamine distribution patterns in rat thalamus

Detailed distribution patterns for norepinephrine and dopamine in rat thalamus were determined by liquid chromatographic assays. The norepinephrine distribution is in good accord with that suggested previously by neuroanatomical tracing techniques. Detailed anterior-posterior concentration profiles for norepinephrine are summarized. An unexpected finding was a strong animal variability in thalamic dopamine content. Thalami were distinguished as having either low, intermediate or extremely high dopamine concentrations.

The occurrence and synthesis of octopamine and catecholamines in the cestode Hymenolepis diminuta

The occurrence and regional distribution of octopamine and catecholamines in the cestode Hymenolepis diminuta were determined by specific radioenzymatic assays. Octopamine was found in the anterior and middle regions (2630 ± 490 and 520 ± 290 pg/mg protein, respectively) but not in the posterior region. On the other hand, dopamine was detected in the anterior region only (4.65 ± 1.42 pg/mg protein). Adrenalin and noradrenalin were not detected in any of the regions. The ability of intact H. diminuta to synthesize octopamine, dopamine and noradrenalin was investigated by incubating the worms in vitro in radiolabelled precursors, followed by separation of the 3H-labelled products by thin-layer chromatography. Only traces of [ 3H]dopamine and [ 3H]noradrenalin were produced in the tissues following incubation in [ 3H]tyrosine or [ 3H]tyramine. However, substantial amounts of [ 3H]octopamine were synthesized from both radiolabelled tyrosine and tyramine. These findings are discussed in the light of octopamine as a possible neurotransmitter/neuromodulator in H. diminuta.

A modification of the glyoxylic acid induced histofluorescence technique for demonstration of catecholamines and serotonin in tissues of Aplysia californica

A modified glyoxylic acid technique was used to examine central and peripheral nervous tissues in Aplysia californica. In addition to confirming the distribution of catecholamines and serotonin in the central nervous system, the method demonstrated the presence of monoamines in the opaline gland and bag cell clusters where they may act as transmitters. In conjunction with electrophysiological techniques this method may be useful to identify other monoamine-containing neurons in Aplysia.

Regional distribution of catecholamines in nucleus accumbens of the rabbit.

The nucleus accumbens is an important telencephalic region, which is the target limbic and mesolimbic pathways. Because of an ongoing physiological study of the effects of dopamine, we wanted to determine regional differences of dopamine and norepinephrine concentrations in the nucleus. As determined by radioenzymatic assays, dopamine levels were not significantly different in the anterior-posterior dimension, averaging approximately 187 ng dopamine/mg protein. Substantial amounts of norepinephrine were found throughout the nucleus, but the levels were significantly higher in the caudal portions of the nucleus, being approximately 4.5 times higher than in the anterior portions.

Absence of dopamine-β-hydroxylase in some catecholamine-containing sympathetic ganglion cells of the dog: evidence for dopaminergic autonomic neurones

Previous pharmacological and biochemical findings have suggested the presence of dopaminergic vasomotor neurones in the sympathetic outflows to the kidney and the digits of the dog. In the present study, we have used histochemical techniques to compare the distributions of catecholamines and dopamine-β-hydroxylase-like immunoreactivity in ganglion cell bodies of the sympathetic chain. The results indicate that some neurones in the ganglia supplying the kidney and the hindlimb store a catecholamine but lack dopamine-β-hydroxylase. This finding supports the existence of dopaminergic sympathetic neurones.

Catecholamines and serotonin in the caudal medulla of the rat: Combined neurochemical-histofluorescence study

The distribution of monoamine transmitters among the nuclei of the caudal medulla was determined through the combined use of the Falck-Hillarp histofluorescence method for cellular localization of catecholamines and serotonin and quantitative micropunch neurochemical assays of norepinephrine (NE), dopamine (DA) and 5-hydroxytryptamine (5-HT) content using high performance liquid chromatography with electrochemical detection. These combined methods permit a direct correlation between neurotransmitter levels and fluorescent fiber and perikaryal profiles. Eight medullary nuclei were sampled: 1. (1) dorsal motor nucleus of X and ventral nucleus solitarius. 2. (2) the dorsomedial reticular formation. 3. (3) the hypoglossal nucleus. 4. (4) the ventromedial reticular formation. 5. (5) the nucleus ambiguus. 6. (6) nuclei raphe obscurus and pallidus. 7. (7) the inferior olivary nucleus. 8. (8) the descending (spinal) nucleus of V. Micropunched regions containing neurons which contribute projections to or receive sensory input from the vagus nerve were found to contain relatively high levels of NE, DA and 5-HT, consistent with the high density of catecholamine and 5-HT containing terminals observed in these nuclei. The dorsal motor nucleus of X and ventral nucleus solitarius contained the highest levels of NE, DA and 5-HT (218 ± 10, 31 ± 2 and 75 ± 8 pmole/ mg protein, respectively), whereas the lowest of these amines were found in the descending (spinal) nucleus of V (28 ± 1, 3.6 ± 0.8 and 32 ± 2 pmole/ mg protein, respectively). In general the distribution of catecholamines and 5-HT determined by micropunch/microchemical assay agrees well with the distribution of monoamine terminals detected by fluorescence histochemical techniques.

Increased dopamine in the failing hamster heart: Transvesicular transport of dopamine limits the rate of norepinephrine synthesis

An earlier study demonstrated an increase in dopamine and a decrease in norepinephrine in the failing myopathic hamster heart. There was evidence to suggest that this abnormal distribution of catecholamines was secondary to a marked increase in cardiac sympathetic nerve traffic rather than a peculiarity of hamster cardiomyopathy. In this study the hamster model was used to dissect further the step limiting the conversion of dopamine into norepinephrine. In heart failure, cardiac norepinephrine was reduced from a mean value (± standard error of the mean) of 1,192 ± 176 to 441 ± 61 ng/g (probability [p] < 0.005) and cardiac dopamine was increased from 51 ± 5 to 458 ± 70 ng/g (p < 0.001). Cardiac decompensation also induced increases in both cardiac tyrosine hydroxylase (84 percent, p < 0.001) and cardiac dopamine-beta-hydroxylase (46 percent, p < 0.001) relative to values In the control hearts. In preparations of noradrenergic granules 44 percent of norepinephrine stores and 33 percent of dopamine stores were associated with the vesicular pellet of normal hearts. Congestive heart failure led to a significant decrease in norepinephrine, particularly in the intravesicular compartment, whereas dopamine exhibited an approximately 20-fold rise, reflected exclusively by the supernatant (extravesicular) fraction. Ganglionic blockade restored catecholamine content and distribution to normal. Inhibition of monoamine oxidase doubled the norepinephrine content of failing hearts (p < 0.01) in the absence of a change in dopamine. Catecholamine histofluorescence observations revealed a marked reduction in the number of nerves in apposition to the muscle of myopathic hearts In addition to a sprouting of new adrenergic fibers into connective tissue and in the lipofuscin pigment debris. It is concluded that an increase in sympathetic activity, at least in the myopathie hearts, can lead to a shift in the rate-limiting step for norepinephrine synthesis from the hydroxylation of tyrosine to the transport of dopamine across the membrane of the noradrenergic granule. Furthermore, it appears that one or more deaminated metabolites of dopamine in the extravesicular space contribute to this shift by inhibiting the dopamine uptake system.

Non-neurogenic mechanism for reserpine-induced release of catecholamines from the adrenal medulla of neonatal rats: Possible modulation by opiate receptors

Neonatal rats do not have functional splanchnic nerve connections to the adrenal medulla until approximately one week of postnatal age, yet they are able to respond to some drugs or stresses by releasing adrenal catecholamines. Reserpine (5 mg/kg s.c.) resulted in significant loss (20–40%) of neonatal catecholamines within 4 h; unlike the acute effects of reserpine in the adult, depletion was not prevented by pretreatment with a nicotinic blocking agent, demonstrating that the effect in the neonate is non-neurogenic. Depletion did not result simply from inhibition of catecholamine storage by reserpine, but rather, the non-neurogenic depletion represented net movement of the amines from the storage granules into the extracellular space, as evaluated by the subcellular distribution of catecholamines. These results suggest that the non-neurogenic mechanism represents release of catecholamines. The immature response mechanism disappeared by 11 days of age and was replaced by a completely neurogenic release, demonstrating that the special mechanism is lost within several days of the onset of functional innervation of the adrenal. Experiments also were carried out to test whether endogenous opioids and/or opiate receptors are involved in the non-neurogenic mechanism. Naloxone (5 mg/kg s.c.) potentiated the depletion of catecholamines by reserpine, while methadone (2.5 mg/kg s.c.) inhibited the non-neurogenic response. In contrast, no potentiation of release by naloxone or inhibition by methadone was seen in adult rats. Thus, opiate receptors may modulate only the immature secretory mechanism.

Catecholamine-containing neurons in cultures of fetal rat hypothalamus: distribution, morphology, and maturation.

The distribution, morphology, and maturation of catecholamine (CA) neurons have been studied in hypothalamic explants from late-gestation rats. CA-containing neurons were identified using the glyoxylic acid technique. CA-containing processes were present from all hypothalamic areas except the preoptic region. Several fiber types were identified. CA neurons in vitro resemble CA neurons in adult hypothalamus. This tissue culture system may be useful in the study of a number of properties of hypothalamus CA neurons.

A Histochemical Study of the Gas Gland Innervation in the Atlantic God,Gadus morhua

A histochemical study was made of the distribution of catecholamines and cholinesterases in two autonomic ganglia closely associated with the swimbladder of the Atlantic cod, Gadus morhua. The “swimbladder nerve ganglion” comprised large (40 μm) neurones, the majority of which are positive for both catecholamines and acetylcholinesterase. It is argued that these neurones are mainly adrenergic postganglionic elements of sympathetic pathways which pass through the vago-sympathetic trunk. The “gas gland ganglion” comprised small (20 μm) neurones, positive for acetylcholinesterase but showing no catecholamine reaction. It is argued that these neurones are cholinergic postganglionic elements of the parasympathetic vagal innervation of gas gland cells.