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PubMed1979

Catecholamine and 5-hydroxytryptamine synthesis and metabolism following intracerebroventricular injection of dibutyryl cyclic AMP.

Two to seven days after bilateral implantation of polyethylene tubings into the lateral ventricles rats were injected intracerebroventricularly with dibutyryl cAMP or dibutyryl cGMP. The accumulation of dopa and 5-hydroxytryptophan after inhibition of the aromatic amino acid decarboxylase with 3-hydroxybenzylhyrazine HCl, served as a measure of catecholamine and 5-hydroxytryptamine synthesis in vivo. A dose of 100 microgram dibutyryl cAMP per rat but not dibutyryl cGMP increased the accumulation of dopa in all brain regions by 70 to 130%. Both nucleotides stimulated the formation of 5-hydroxytryptophan in the dopamine-rich part of the limbic system and diencephalon and in addition dibutyryl cAMP increased 5-hydroxytryptophan in C. striatum and cerebellum. Dibutyryl cAMP (25-200 microgram per rat) did not change the dopamine levels in the dopamine-rich parts of the brain but decreased the noradrenaline level of the C. striatum in a dose-dependent manner. The same doses of dibutyryl cAMP elevated the 5-hydroxytryptamine level in brain stem and the level of 5-hydroxyindole acetic acid in diencephalon and the limbic system. The disappearance of noradrenaline but not of dopamine after inhibition of catecholamine synthesis with alpha-methyl-p-tyrosine methylester HCl was accelerated in most brain regions. The data are compatible with the view that dibutyryl cAMP stimulates tyrosine and tryptophan hydroxylase directly. In addition, dibutyryl cAMP appears to enhance the utilization of noradrenaline but not that of dopamine. The increased utilization of 5-hydroxytryptamine may be restricted to the diencephalon and the limbic system.

5-Hydroxytryptophan↗
PubMed2005

Striatal dopamine transporter availability with [123I]beta-CIT SPECT is unrelated to gender or menstrual cycle.

OBJECTIVES: The effect of gender and female menstrual cycle on human striatal dopamine transporters (DATs) was investigated with single-photon emission computed tomography (SPECT) using the ligand 2beta-carbomethoxy-3beta-(4-[(123)I]iodophenyl)tropane. METHODS: Ten female subjects aged 18-40 years (25.3+/-7.3 years) were scanned twice during the early follicular and the mid-luteal phases to detect any hormone-mediated changes in DAT availability in the striatum or serotonin transporter (SERT) availability in brainstem-diencephalon. Plasma estradiol and progesterone levels were obtained at the time of SPECT and confirmed the expected increases from the follicular to the luteal phases. Finally, in a post hoc analysis of a previously published healthy-subject sample, striatal DAT availability was compared between 70 male and 52 female subjects who ranged in age from 18 to 88 years. RESULTS: In the ten menstrual cycle subjects, DAT availability (V(3)'') in striatum and SERT availability in brainstem-diencephalon did not differ between follicular and luteal phases. Moreover, change in V(3)'' for striatum or brainstem-diencephalon was uncorrelated with change in plasma estradiol or progesterone from the follicular to the luteal phase. In the larger healthy-subject sample, there was no significant effect of gender or the interaction of age and gender on striatal V(3)''. CONCLUSIONS: These findings suggest that in using DAT or SERT ligands in the study of neuropsychiatric disorders, matching of female subjects according to a menstrual cycle phase is unnecessary. Although the present investigation did not confirm previous reports of gender differences in striatal DAT availability, controlling for gender in such studies still seems advisable.

Adolescent↗
PubMed1998

Astroglial architecture of the carp (Cyprinus carpio) brain as revealed by immunohistochemical staining against glial fibrillary acidic protein (GFAP).

The present paper is the first comprehensive study on the astroglia of a teleost fish that is based on the immunohistochemical staining of GFAP (glial fibrillary acidic protein, an immunohistochemical marker of astroglia). The ray-finned fishes (Actinopterygii) and their largest group, the Teleostei, represent a separate pathway of vertebrate evolution. Their brain has a very complex macroscopic structure; several parts either have no equivalents in tetrapods or have a very different shape, e.g., the telencephalon. The results show that the teleost brain has a varied and highly specialized astroglial architecture. The primary system is made up of radial glia, which are of ependymal origin and cover the pial surface with endfeet. The tendency is, however, that the more caudal a brain area is, the less regular is the radial arrangement. A typical radial glia dominates some parts of the diencephalon (median eminence, lobus inferior and habenula) and the telencephalon. In the rest of the diencephalon and in the mesencephalon, the course of the glial fibers is modified by brain tracts. The most specialized areas of the teleost brain, the optic tectum and the cerebellum, display elaborate variations of the original radial system, which is adapted to their layered organization. In the cerebellum, an equivalent of the Bergmannglia can be found, although its fiber arrangement shows meaningful differences from that of mammals or birds. In the lower brain stem radial glia are confined to fibers separating the brain tracts and forming the midline raphe. A dense ependymoglial plexus covers the inner surface of the tectum and the bottom of the rhombencephalic ventricle, intruding into the vagal and facial lobes. The structure and the position of the rhombencephalic plexus suggest that it corresponds to a circumventricular organ that entirely occupies the bottom of the ventricle. Perivascular glia show an unusual form as they consist of long fibers running along the blood vessels. In the large brain tracts long glial fibers run parallel with the course of the neural fibers. At least in the diencephalon, these glial fibers seem to be modified radial fibers. Real astrocytes (i.e., stellate-shaped cells) can be found only in the brain stem and even there only rarely. The glial specialization in the various areas of the teleost brain seems to be more elaborate than that found either in amphibia or in reptiles.

Animals↗
PubMed1977

Ascending projections of the locus coeruleus in the rat. I. Axonal transport in central noradrenaline neurons.

Axonal transport of protein and metabolites of L-[3H(G)]3, 4-dihydroxyphenylalanine ([3H]DOPA) was studied in the central noradrenaline neurons of the pontine nucleus locus coeruleus and was correlated with regional alterations of noradrenaline content following destruction of the nucleus. Unilateral lesions of the locus coeruleus produce a partial depletion of noradrenaline in the ipsilateral hypothalamus and telencephalon, indicating that these neurons project widely to the ipsilateral forebrain. Twenty-four to 48 h following local injections of 50 micronCi [3H]proline, locus coeruleus neurons take up labeled material and transport it, presumably as protein, to ipsilateral structures in the midbrain, diencephalon and telencephalon including the neocortex. Similarly 8 h after injection of 25 micronCi [3H]DOPA into the locus coeruleus, transport of material including catecholamines occurs to ipsilateral diencephalon and telencephalon. Axonal transport of proteins to telencephalic structures is greatly diminished by selective lesions of catecholamine terminals with 6-hydroxydopamine (6-OHDA) and following destruction of the medial forebrain bundle. These results provide further support for the view that noradrenaline neurons of the locus coeruleus nucleus project widely within the neuraxis to ipsilateral structures of the brain stem, diencephalon and telencephalon, including all cortical areas. In addition, evidence is presented for a contralateral projection with a similar distribution. The rate of axonal transport of labeled protein and metabolites of [3H]DOPA including [3H]catecholamines in central noradrenaline neurons is estimated to be 3-4 mm/h and is accordingly similar to that reported for noradrenaline neurons of the peripheral sympathetic nervous system.

Adrenergic Fibers↗
PubMed1988

Decrease in tetrahydrobiopterin content and neurotransmitter amine biosynthesis in rat brain by an inhibitor of guanosine triphosphate cyclohydrolase.

To investigate the regulatory role of tetrahydrobiopterin in neurotransmitter amine biosynthesis, 2,4-diamino-6-hydroxypyrimidine, a potent inhibitor of guanosine triphosphate cyclohydrolase which is a rate-limiting enzyme of tetrahydrobiopterin biosynthesis, was administered intraperitoneally to weanling rats. Four h after 4 injections at 4-h intervals, the biopterin contents in plasma and liver were reduced to the level of 9 and 3.5%, respectively, of those in the control group injected with saline; while the contents in the whole brain, neocortex + striatum, diencephalon, and brainstem were 34, 50, 33 and 28%, respectively, of the control level. When in vivo tyrosine and tryptophan hydroxylase activities were measured over a 30-min period after the inhibition of aromatic amino acid decarboxylase, the accumulation of dihydroxyphenylalanine was reduced to 74, 77, 67 and 69% of the control in the whole brain, neocortex + striatum, diencephalon, and brainstem, respectively; and the accumulation of 5-hydroxytryptophan, to 71, 74, 66 and 65% of the control, respectively. On the other hand, 5-hydroxytryptamine and 5-hydroxyindole acetic acid contents were not altered in any brain regions, although norepinephrine and dopamine contents were reduced to approximately 70% of the control in the brainstem and the contents of dopamine metabolites were significantly decreased in the diencephalon and brainstem. Plasma phenylalanine level was significantly elevated, while the plasma tyrosine level was reduced, compared with the control level of these amino acids. These results indicate that the drug-treated rats could be an animal model for tetrahydrobiopterin-deficient disease involving neurological disorder.

5-Hydroxytryptophan↗
PubMed1984

Stereotyped and variable growth of redirected Mauthner axons.

To assay the axon tract organizing capabilities of different regions of the vertebrate CNS, Mauthner axons were redirected by grafting supernumerary hindbrains in Xenopus embryos. The 63 redirected Mauthner axons thus produced included donor axons projecting into the host CNS and host axons that grew through the graft or that were redirected in the host CNS. Two major phenomena were observed. Caudal to the optic chiasm, the Mauthner axons followed a single ipsilateral stereotyped route--the basal substrate pathway--extending in the ventral and ventrolateral marginal zone from the diencephalon to the caudal spinal cord. In contrast, rostral to the optic chiasm, these same Mauthner axons followed variable ipsilateral and contralateral routes. Even pairs of Mauthner axons entering the optic chiasm side-by-side eventually followed different routes in normal forebrains. The contrasting behaviors of the Mauthner axons growing in the rostral diencephalon and telencephalon and of the same Mauthner axons growing elsewhere suggest that there are differences in the effective guidance cues between these two regions of the developing brain. This is consistent with other types of neuroanatomical and neuroembryological evidence indicating a fundamental division between the rostral and the caudal diencephalon.

Animals↗
PubMed1983

Effects of chronic ingestion of alcohol in the pregnant rat on catecholamine-sensitive adenylate cyclase in the brain of mothers and their offspring.

To study the effects of maternal alcohol ingestion on brain adenylate cyclase in offspring, rats were given ethanol in the drinking water before, during and after gestation. Fifteen-day-old pups from alcoholic mothers had reduced body and wet brain weight and protein content with normal concentrations of DNA and RNA in brain; pups from mothers deprived of alcohol from the 21st day of gestation had normal growth and brain protein, DNA, RNA and cyclic nucleotide concentrations. Adenylate cyclase activity was less in homogenates from the diencephalon, mesencephalon and cerebellum of alcoholic and alcohol-deprived mothers than in the corresponding homogenates from control mothers. A reduction of the enzyme activity was only observed in homogenates from diencephalon of offspring from alcoholic and alcohol-deprived mothers as compared with controls. Sensitivity of adenylate cyclase to noradrenaline was reduced in the corpora striata and diencephalon from alcoholic mothers, while these changes were absent in alcohol-deprived mothers and their offspring. Ingestion of ethanol in pregnant mothers does not seem to affect significantly the adenylate system in the brain of the offspring.

Adenylyl Cyclases↗
PubMed1987

Effects of chronic administration of dextroamphetamine on enzymes of energy metabolism in regions of the rat brain.

In the present study the effects of chronic administration of dextroamphetamine on energy metabolism in the brain of the rat were examined. The enzymes studied were: hexokinase (soluble and particulate forms), phosphofructokinase, pyruvate kinase, lactate dehydrogenase, citrate synthase, NAD+ and NADP+-dependent isocitrate dehydrogenases, succinate dehydrogenase and malate dehydrogenase. All the activities of the enzymes were assayed in four regions of the brain of the rat (cerebellum, medulla oblongata and pons, cererbral cortex and diencephalon). Rats were injected intaperitoneally once daily with dextroamphetamine for 20 consecutive days. The initial dose was 5 mg/kg/day and the dose was then increased by 1 mg/kg/every 5 days up to a total of 8 mg/kg/day on days 16-20. In the glycolytic enzymes a reduction of the activity of phosphofructokinase was found in the diencephalon and an increase of the activity of pyruvate kinase and lactate dehydrogenase in the diencephalon and medulla oblongata and pons, respectively. Citrate synthase was the only enzyme in the Krebs' cycle affected by chronic administration of dextroamphetamine. The results presented here show that chronic administration of dextroamphetamine produced important changes in some enzymes of glycolysis and the Krebs' cycle in the brain of the rat.

Animals↗
PubMed1976

Neurochemical correlates of behavior: levels of amino acids in four areas of the brain of the rat during drug-induced behavioral excitation.

The levels of GABA, aspartate, glutamate, glycine and alanine were determined in 4 specific brain areas (telencephalon, diencephalon-mesencephalon cerebellum and pons-medulla oblongata) of rats killed during a period of drug-induced behavioral excitation. Behavioral excitation was obtained in adult, male Wistar rats working on a Sidman shock-avoidance schedule following administration of 2 mg/kg tetrabenazine (TBZ) 18 hr after iproniazid (50 mg/kg) pretreatment. When compared to trained animals (working on the avoidance schedule but receiving no drugs), the excited rats had increased levels of GABA in the telencephalon and diencephalon-mesencephalon, decreased levels of aspartate in all 4 brain areas, and a lower content of glycine in the pons-medulla region. The changes in the levels of aspartate in all areas of the brain, GABA in the diencephalon-mesencephalon, and glycine in the pons-medulla were significantly correlated (p less than 0.01) with the degree of excitation. It was observed that avoidance training alone produced increases in the levels of four amino acids: aspartate in telencephalon and cerebellum, GABA in cerebellum, and glycine and glutamate in the pons-medulla. The injection of iproniazid alone or iproniazid followed by TBZ into naive animals had little effect on the levels of the five amino acids. The data are discussed in terms of aspartate and GABA interacting as neurotransmitters with cholinergic and catecholaminergic and/or serotonergic neurons to produced the behavioral excitation.

Amino Acids↗
PubMed1986

Ontogeny of gamma-melanocyte-stimulating hormone in the brain and hypophysis of the rat: an immunohistochemical analysis.

The ontogeny of gamma-melanocyte stimulating hormone (gamma-MSH)-like immunoreactive (gamma-MSHI) structures in the brain and hypophysis was investigated in the rat by means of indirect immunofluorescence. gamma-MSHI neurons in the arcuate nucleus appeared at Day 13 of gestation, in the anterior hypophysis at Day 16, in the intermediate lobe at Day 18, and in the nucleus commissuralis after birth. gamma-MSHI fibers first appeared at Day 15 of gestation in the hypothalamic area and extended dorsally to reach the surface of the diencephalon. At Day 16 of gestation, another gamma-MSHI fiber bundle was found that ran laterally along the ventral surface of the diencephalon. At Day 19, immunoreactive fibers first appeared in the forebrain, diencephalon, midbrain and upper pons, and thereafter they increased in number, reaching a maximum at postnatal Day 15. gamma-MSHI fibers in the lower pons and medulla oblongata first appeared after birth. They also increased in number with age and reached a maximum at postnatal Day 15. The present ontogenetical study has demonstrated the different times of first appearance of each of the 3 major gamma-MSHI structures-containing cell groups in the brain, i.e., the arcuate nucleus, hypophysis and nucleus commissuralis. These findings may reflect the different functions of 3 groups of cells.

Animals↗
PubMed1992

Plasticity and ontogeny of the central 5-HT transporter: effect of neonatal 5,7-dihydroxytryptamine lesions in the rat.

5,7-Dihydroxytryptamine (5,7-DHT) is unique as a serotonin (5-HT) neurotoxin in that i.p. injection of neonatal rats increases concentrations of 5-HT in brainstem while depleting 5-HT in cortex, hippocampus and spinal cord. To study the mechanism of this effect we measured the 5-HT transporter or uptake site, a presynaptic marker, using [3H]paroxetine binding. There were significant regional differences in Bmax of vehicle-injected rats: brainstem, diencephalon > striatum, cortex, spinal cord > hippocampus, cerebellum. There were also regional differences in the ontogeny of bindings sites: at postnatal day 7, [3H]paroxetine sites were 39% of adult levels in cortex compared to 63% in brainstem. Thirty days after 100 mg/kg 5,7-DHT i.p., Bmax of [3H]paroxetine binding was significantly increased in brainstem (+67%) and diencephalon (+136%), whereas it decreased in cortex (-59%), hippocampus (-94%) and spinal cord (-99%), striatum (-41%) and cerebellum (-37%). KD remained unaltered. In dose-response studies (0-200 mg/kg), 50 mg/kg was the threshold dose for Bmax effects and 200 mg/kg was lethal. In weekly time-course studies, changes were apparent 1 week after 5,7-DHT lesions. Binding site increases in diencephalon and brainstem were not maximal until 3 weeks after injection, whereas percent decreases in cortical sites remained unchanged at each week studied. Lesion effects on the ontogeny of [3H]paroxetine binding sites were region-dependent: cortical sites continued to increase with age but spinal sites did not. There was no significant recovery in spinal cord.(ABSTRACT TRUNCATED AT 250 WORDS)

5,7-Dihydroxytryptamine↗
PubMed1994

Induction of interleukin-1 in various brain regions after peripheral and central injections of lipopolysaccharide.

The presence of bioactive interleukin-1 (IL-1) in various brain regions (cerebellum, cortex, brainstem, diencephalon or hippocampus) after either intraperitoneal (i.p.) or intraventricular (i.c.v.) injection of lipopolysaccharide (LPS) was studied in the rat. To detect IL-1, extracellular fluid and cell lysate were fractionated by gel exclusion chromatography and fractions tested for thymocyte stimulation; presence of IL-1 was confirmed by blockade of stimulation by addition to the assay of a monoclonal antibody (mAb) to IL-1 receptor. When LPS was infused i.c.v., IL-1 was detected in the brainstem and diencephalon 2 h after injection, and in all the brain regions except cerebellum 6 h after injection; IL-1 was not detected in the plasma of these animals. When LPS was injected i.p., IL-1 was detected in the plasma but not in the brain 2 h after the injection, and in all brain regions but not in the plasma 6 h after the injection. In all of these cases, IL-1 was found in extracellular fluid; in some cases (cortex, cerebellum) cell lysate of the region did not produce detectable bioactivity, thereby indicating that IL-1 in these brain regions is processed to active peptide during release, as has been reported in the periphery. In those cases where bioactivity was detected in cell lysate (brainstem, diencephalon), bioactivity was not blocked by IL-1 receptor mAb, indicating presence of a non-IL-1 stimulating factor. These results further support the idea that IL-1 is secreted by cells in the brain, and indicate that it is found in the extracellular fluid of many brain regions following an appropriate stimulus in the periphery as well as in the brain.

Animals↗
PubMed1990

Amino acid content in several brain regions of the active and hibernating frog, Rana esculenta.

1. Total free amino acid contents in the optic lobe and diencephalon increased significantly during hibernation. 2. Free glutamate + glutamine showed significant increases in the cerebral hemisphere, optic lobe, medulla oblongata and diencephalon. 3. Free aspartate + asparagine showed significant increases in the cerebral hemisphere, optic lobe, diencephalon and olfactory lobe. 4. GABA showed a significant change only in the medulla oblongata. 5. Total protein amino acid level in the cerebellum and olfactory lobe decreased significantly during hibernation and most of the amino acids decreased significantly in these regions. 6. The amino acid metabolism during amphibian hibernation differs from that of the mammal.

Amino Acids↗
PubMed1984

The effect of long-term ethanol maternal ingestion and withdrawal on brain regional monoamine and amino acid precursors in 15-day-old rats.

The effects of alcohol on brain monoamines were studied in 15-day-old offspring of rats given ethanol until the 21st day of gestation or the 15th day post partum. Increased noradrenaline concentrations were found in limbic system, hemispheres, diencephalon and brain stem of pups from mothers under alcohol treatment, and in hemispheres, diencephalon and brain stem of pups from deprived mothers. Serotonin and 5-hydroxyindol acetic acid were augmented in limbic system, diencephalon and brain stem of pups from mothers under alcohol whereas tyrosine was decreased in all brain regions studied in pups from alcoholic and deprived mothers. Consequently, chronic ethanol ingestion by pregnant rats has deleterious effects on CNS development of 15-day-old offspring which persists 15 days after alcohol withdrawal.

Alcoholism↗
PubMed1983

Development of the serotonergic system in the rat embryo: an immunocytochemical study.

The development of central serotonergic neurons has been examined immunocytochemically utilizing an antiserum to serotonin (5-HT). Cells of the B4-B9 complex are first detected early on embryonic day 13 (E13; 7 mm crown rump length, CRL) and increase rapidly in number through E15 when they appear as bilateral columns situated from just caudal to the mesencephalic flexure to the pontine flexure. Aggregation of cells into subgroups is apparent soon after 5-HT neurons leave the ventricular zone, allowing the identification of certain subdivisions of the B4-B9 complex long before they assume their adult locations. The initial detection of 5-HT immunoreactive cells in the medulla occurs 1-2 days after the appearance of cells in the B4-B9 complex, although it has been reported that the time of origin of medullary raphe neurons (B1-B3) occurs before that of raphe neurons in the midbrain and pons (B4-B9). The first medullary 5-HT neurons, comprising the B3 subdivision occur ventro-laterally on E14 (10-11 mm CRL) at least 1-2 days before midline 5-HT neurons are visualized in the B1 and B2 groups. Thus, in contrast to cells in the B4-B9 complex, medullary 5-HT neurons complete much of their migration before they can be detected immunocytochemically, indicating that the time of onset of transmitter synthesis and storage may differ during differentiation of cells sharing a common neurotransmitter phenotype. The formation of ascending 5-HT fiber projections occurs rapidly from cells of the B4-B9 complex. Within 24 hours after the initial detection of 5-HT fiber immunoreactivity in such cells at E13, their axons are seen entering the caudal diencephalon (E14). These fibers have traversed the diencephalon and floor of the telencephalon by E15-E16 and reach the frontal neocortical pole by E17. The main ascending bundle of 5-HT axons courses through the diencephalon in the vicinity of the medial forebrain bundle, although some fibers also diverge and travel along certain pre-existing non-5HT pathways. However, examples are also found of acute directional changes in 5-HT fiber growth which do not appear to be associated with pre-formed non-5HT pathways. The pattern of ascending fiber outgrowth suggests a priority routing system which provides certain regions with 5-HT axons in a preferential sequence irrespective of the distance of these areas from 5-HT cell groups or from major bundles of ascending 5-HT fibers.

Afferent Pathways↗
PubMed1988

Serotonin uptake in the central nervous system of rats fed a corn-diet.

1. Endogenous serotonin (5-HT), 5-hydroxyindol acetic acid (5-HIAA) content and exogenous 5-HT uptake (Km and Vmax) were measured in different brain regions (cerebellum, diencephalon, brain stem and telencephalon) of rats fed with a corn diet and restricted protein (8%) diet during 6 weeks. 2. A reduction of 5-HT levels was found in all regions studied of animals fed a corn diet, whereas, 5-HIAA was only decreased in brain stem and diencephalon. 3. An important increase in Km and Vmax were registered in brain stem and diencephalon of protein restricted animals, whereas, an increase of 5-HT uptake affinity in cerebellum, brain stem and telencephalon (35, 42 and 33% respectively) was observed. Simultaneously, under corn diet conditions, the Vmax decreased 40, 30 and 34% respectively in those regions. 4. It is suggested that the brain stem was the more sensitive area under nutritional restricted conditions and the development of some possible compensatory mechanisms of the 5-HTergic system is discussed.

Animals↗
PubMed1993

Navigational errors made by growth cones without filopodia in the embryonic Xenopus brain.

We have developed an exposed brain preparation for observing growth cone pathfinding behavior while performing in vivo pharmacological manipulations, and we used it to test whether Xenopus retinal growth cones need filopodia to navigate. Time-lapse video observation showed that cytochalasin B acted quickly and reversibly when applied; cytochalasin B-treated growth cones lacked filopodia, but had active lamellipodia and continued to advance slowly. Whereas normal retinotectal axons visualized with horseradish peroxidase turn caudally in the mid-diencephalon to reach the tectum, cytochalasin B-treated axons grew past the normal turning point and, instead, continued straight within the diencephalon. In dose-response experiments, pathfinding became abnormal in the same concentration range in which filopodia disappeared. These results suggest that filopodia are necessary for retinal growth cones to respond to guidance signals in the diencephalon.

Animals↗
PubMed2004

Pituitary adenylate cyclase-activating polypeptide in the brain, spinal cord and sensory organs of the zebrafish, Danio rerio, during development.

The distribution of Pituitary adenylate cyclase-activating polypeptide (PACAP) was investigated in the brain, pituitary and sensory organs of the zebrafish, Danio rerio, during development, in juvenile and adult specimens, using the immunofluorescence method. In 24 h post fertilization (hpf) embryos, PACAP immunoreactive cells appeared in the rostral telencephalon, dorsal diencephalon, caudal and medial rhombencephalon, spinal cord and retina. At 48 hpf stage, positive cells were present in the dorsal diencephalon, medial rhombencephalon, spinal cord, retina and olfactory placode (Op). At 72 hpf stage, additional immunoreactive elements appeared in the medial telencephalon, hypothalamus, mesencephalic tegmentum, retina and otic sensory epithelium (Ose). At day 5, new immunoreactive cells were found in the anterior rhombencephalon and pituitary pars distalis. At day 13, positive cells were mainly concentrated in the mesencephalic tegmentum and spinal cord. In the telencephalon, diencephalon, rhombencephalon and pituitary, the distribution of positive cells was similar to that previously reported. At 1 month stage, positive cells were detected in the hypothalamus, nucleus of the medial longitudinal fascicle (nMlf), rhombencephalic griseum centrale (Gc) and pituitary pars distalis. At 2-3 month stages, immunoreactive elements were found in several hypothalamic nuclei, in the mesencephalic nucleus isthmi, cerebellum and pituitary. In adults, PACAP immunoreactivity was confined to a few brain regions and the pituitary. PACAP immunoreactivity was transiently expressed in several regions suggesting that the peptide may have a role in the control of cells differentiation and proliferation during zebrafish ontogeny. The finding of positive fibers in the pituitary from day 5 onward indicates that PACAP may function from this stage as a hypophysiotropic peptide.

Age Factors↗