RESUMO
The satiety-promoting action of oleoylethanolamide (OEA) has been associated to the indirect activation of selected brain areas, such as the nucleus of the solitary tract (NST) in the brainstem and the tuberomammillary (TMN) and paraventricular (PVN) nuclei in the hypothalamus, where noradrenergic, histaminergic and oxytocinergic neurons play a necessary role. Visceral ascending fibers were hypothesized to mediate such effects. However, our previous findings demonstrated that the hypophagic action of peripherally administered OEA does not require intact vagal afferents and is associated to a strong activation of the area postrema (AP). Therefore, we hypothesized that OEA may exert its central effects through the direct activation of this circumventricular organ. To test this hypothesis, we subjected rats to the surgical ablation of the AP (APX rats) and evaluated the effects of OEA (10mgkg-1 i.p.) on food intake, Fos expression, hypothalamic oxytocin (OXY) immunoreactivity and on the expression of dopamine beta hydroxylase (DBH) in the brainstem and hypothalamus. We found that the AP lesion completely prevented OEA's behavioral and neurochemical effects in the brainstem and the hypothalamus. Moreover OEA increased DBH expression in AP and NST neurons of SHAM rats while the effect in the NST was absent in APX rats, thus suggesting the possible involvement of noradrenergic AP neurons. These results support the hypothesis of a necessary role of the AP in mediating OEA's central effects that sustain its pro-satiety action.
Assuntos
Área Postrema/efeitos dos fármacos , Tronco Encefálico/efeitos dos fármacos , Ingestão de Alimentos/efeitos dos fármacos , Endocanabinoides/farmacologia , Hipotálamo/efeitos dos fármacos , Ácidos Oleicos/farmacologia , Animais , Área Postrema/fisiologia , Tronco Encefálico/fisiologia , Dopamina beta-Hidroxilase/análise , Dopamina beta-Hidroxilase/metabolismo , Hipotálamo/fisiologia , Masculino , Ocitocina/análise , Ocitocina/metabolismo , PPAR alfa/análise , PPAR alfa/metabolismo , Proteínas Proto-Oncogênicas c-fos/análise , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos WistarRESUMO
The aim of the study was to explore whether the biphasic time course of the vasospastic response following experimental subarachnoid hemorrhage is associated with any concomitant changes in the amount of cerebral dopamine beta-hydroxylase in the noradrenergic central nervous system. A single-hemorrhage animal model was used. Rabbits were sacrificed from day 1 to day 8 after subarachnoid hemorrhage. Intimal corrugation of the basilar artery and the amount of cerebral dopamine beta-hydroxylase in the hypothalamus and brain stem were measured each day. Vasospastic changes occurred in the biphasic manner following subarachnoid hemorrhage. More profound vasospastic corrugation occurred in the acute phase, followed by a slightly less intense corrugation in the chronic phase (between days 5 and 8 after the subarachnoid hemorrhage). Simultaneously, a clear concomitant biphasic time course developed in the form of an increased amount of dopamine-beta-hydroxylase in the noradrenergic nervous system of the rabbit hypothalamus and brain stem during the acute and chronic phases after the subarachnoid hemorrhage. Statistically significant correlation between basilar artery corrugation and the amount of dopamine beta-hydroxylase was found. These results suggest the possible role of the central sympathetic system in the pathogenesis of vasospasm. At the same time, this study demonstrates the chronological similarity of the vasospastic development after subarachnoid hemorrhage in the animal experimental model with the human time course of vasospasm.
Assuntos
Fibras Adrenérgicas/enzimologia , Artéria Basilar/inervação , Tronco Encefálico/enzimologia , Dopamina beta-Hidroxilase/análise , Hipotálamo/enzimologia , Proteínas do Tecido Nervoso/análise , Hemorragia Subaracnóidea/complicações , Vasoespasmo Intracraniano/fisiopatologia , Doença Aguda , Fibras Adrenérgicas/fisiologia , Animais , Artéria Basilar/patologia , Sangue , Doença Crônica , Cisterna Magna , Modelos Animais de Doenças , Feminino , Injeções , Coelhos , Fatores de Tempo , Túnica Íntima/patologia , Vasoespasmo Intracraniano/enzimologia , Vasoespasmo Intracraniano/etiologiaRESUMO
BACKGROUND: The aim of this study of experimental subarachnoid haemorrhage (SAH) and exclusion of the sympathetic nervous system (SNS) in rabbits was to find out if changes in the central noradrenergic areas of the hypothalamus and brain stem could be ascertained, in parallel with measurement of the intensity of chronic cerebral vasospasm in the basilar arteries. METHODS: Histologic specimens were prepared by perfusion fixation on day 8 after the SAH. The spastic effect of experimentally induced SAH in New Zealand rabbits was investigated: firstly, using our previously developed method for measuring the corrugation coefficient (CC) of the vessel intima on precisely defined locations of the basilar artery (BA) with the aid of computer image analysis; and secondly, by immunohistochemical assessment of the concentration and localization of dopamine beta-hydroxylase (DBH), using anti-DBH, at precisely defined sites of the hypothalamus and brain stem of the same rabbit. RESULTS: The intima of the BA, assessed by CC, was significantly less corrugated and had significantly less DBH in group A (the control group without SAH and without additional interventions; mean CC = 1.192, P = 0.004; median DBH = 0.50, P = 0.044), in group C (SAH and alpha-blocker phenoxybenzamine; mean CC = 1.142, P = 0.000; median DBH = 0.75, P = 0.001), and in group D (SAH and cervical gangliectomy; mean CC = 1.210, P = 0.003; median DBH = 0.50, P = 0.002) compared with group B (rabbits with SAH and without medication). Group B showed a significantly more intensive accumulation of DBH (median DBH = 1.15) and, according to the CC (mean CC = 1.369), more intensive corrugation of the intima of BA than all other groups. The correlation between CC and DBH for all the rabbits (groups A, B, C and D together) was significantly positive (Spearman Rho = 0.470; p = 0.010). CONCLUSIONS: The results of this study demonstrated: firstly, an intensive excitatory influence of SAH on the quantity of DBH in central noradrenergic areas in the hypothalamus and brain stem; secondly, a very effective influence of peripheral and systemic sympathetic exclusion on lowering the quantity of central sympathetic DBH; thirdly, that the changes in the BA of individual rabbits occur simultaneously with corresponding changes in DBH-containing neurons, thus suggesting the likelihood of SNS involvement in the pathogenesis of post-SAH vasospasm in rabbits.
Assuntos
Encéfalo/enzimologia , Dopamina beta-Hidroxilase/metabolismo , Hemorragia Subaracnóidea/complicações , Sistema Nervoso Simpático/fisiopatologia , Vasoespasmo Intracraniano/etiologia , Animais , Artéria Basilar/patologia , Artéria Basilar/fisiopatologia , Encéfalo/patologia , Tronco Encefálico/patologia , Tronco Encefálico/fisiologia , Tronco Encefálico/fisiopatologia , Doença Crônica , Modelos Animais de Doenças , Dopamina beta-Hidroxilase/análise , Feminino , Técnicas Histológicas , Hipotálamo/patologia , Hipotálamo/fisiologia , Hipotálamo/fisiopatologia , Imuno-Histoquímica , Coelhos , Hemorragia Subaracnóidea/patologia , Hemorragia Subaracnóidea/fisiopatologia , Sistema Nervoso Simpático/fisiologia , Fatores de Tempo , Túnica Íntima/patologia , Vasoespasmo Intracraniano/patologia , Vasoespasmo Intracraniano/fisiopatologiaRESUMO
Chronic frontal lobe functional deficits after traumatic brain injury (TBI) may be associated with altered catecholamine systems in the frontal cortex. To test this, tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) levels were examined by immunohistochemistry and Western blot at 1, 7, 14, and 28 days after TBI or sham surgery. No alterations in DBH levels were observed by Western blot at any time point examined, but there was a significant increase in TH expression 28 days after TBI (optical density 334 +/- 68% or 3.3-fold, ipsilateral and 218 +/- 39% or 2.2-fold, contralateral) relative to the sham controls. The increase in TH may reflect a compensatory response of dopaminergic neurons to upregulate their synthesizing capacity and increase the efficiency of dopamine neurotransmission chronically after TBI.
Assuntos
Lesões Encefálicas/metabolismo , Dopamina beta-Hidroxilase/metabolismo , Lobo Frontal/enzimologia , Lobo Frontal/lesões , Tirosina 3-Mono-Oxigenase/metabolismo , Animais , Western Blotting , Dopamina beta-Hidroxilase/análise , Lobo Frontal/química , Imuno-Histoquímica , Masculino , Ratos , Tirosina 3-Mono-Oxigenase/análiseRESUMO
Previous studies have shown that gonadectomy in adult male rats induces a complex series of region- and time-specific changes in the density of presumed cerebral cortical dopamine axons that are immunoreactive for tyrosine hydroxylase. The present study asked whether noradrenergic cortical afferents also show hormone sensitivity by assaying axons immunoreactive for the enzyme dopamine-beta-hydroxylase in representative areas of acutely and chronically gonadectomized and sham-operated adult male rats. Catecholamine afferents (both tyrosine hydroxylase-immunoreactive and dopamine-beta-hydroxylase-immunoreactive) were also quantified in gonadectomized rats supplemented with testosterone propionate, with 17-beta-estradiol, or with 5-alpha-dihydrotestosterone. Analyses of noradrenergic (dopamine-beta-hydroxylase) afferents revealed no differences in axon appearance or density among the hormonally intact and hormonally manipulated groups. However, analyses of tyrosine hydroxylase immunoreactivity revealed an unexpected division of labor among ovarian and testicular hormones in ameliorating the effects of acute verses chronic hormone deprivation on these afferents. Estradiol replacement attenuated the decreases in immunoreactivity induced by acute gonadectomy, but was ineffective in suppressing changes in immunoreactivity stimulated by chronic gonadectomy. In contrast, supplementing gonadectomized animals with dihydrotestosterone provided no protection from acute decreases in innervation, but fully attenuated both the supragranular decreases and infragranular increases in tyrosine hydroxylase-immunoreactive axon density that mark the association cortices of chronically gonadectomized rats. Together these findings indicate both long- and short-term effects of gonadectomy on cortical catecholamines, principally target dopamine afferents, and that chronic gonadectomy, which selectively disturbs dopamine innervation in the prefrontal cortices, involves a compromise in androgen signaling pathways.
Assuntos
Axônios/química , Córtex Cerebral/química , Dopamina beta-Hidroxilase/análise , Orquiectomia , Tirosina 3-Mono-Oxigenase/análise , Vias Aferentes/química , Animais , Axônios/efeitos dos fármacos , Catecolaminas/análise , Córtex Cerebral/efeitos dos fármacos , Di-Hidrotestosterona/sangue , Di-Hidrotestosterona/farmacologia , Dopamina beta-Hidroxilase/efeitos dos fármacos , Estradiol/sangue , Estradiol/farmacologia , Hormônios Esteroides Gonadais/sangue , Hormônios Esteroides Gonadais/farmacologia , Gônadas/química , Masculino , Orquiectomia/métodos , Ratos , Ratos Sprague-Dawley , Testosterona/sangue , Testosterona/farmacologia , Tirosina 3-Mono-Oxigenase/efeitos dos fármacosRESUMO
The neuropeptide galanin (Gal) is found throughout the central nervous system. Of particular interest is the fact that Gal is present within the majority of noradrenergic locus coeruleus (LC) neurons. However, very few, if any, Gal-immunoreactive fibers have been identified in many of the major efferent targets of LC, including sensory neocortex and dorsal thalamus. The goal of the present study was to examine the Gal fiber innervation of the rodent trigeminal somatosensory system and its connection to the LC. Our results show that at least two different morphological profiles of Gal-immunoreactive fibers are present within relay nuclei along the ascending trigeminal pathway. Numerous small caliber Gal-immunoreactive fibers with bouton-like swellings were noted within the barrel cortex, the ventroposterior medial (VPM) nucleus, the posterior medial (POm) nucleus, the zona incerta (ZI), the reticular nucleus (nRT) of the thalamus, and the principal (PrV) and spinal (SpV) nuclei of the trigeminal complex. Immunoreactive fibers were prevalent in, but not restricted to, layer I of the barrel cortex. Within the somatosensory thalamus, the density of Gal-immunoreactive fibers was higher in POm than in VPM. Laminae I and II of SpV and the nRT and ZI also contained dense, large-diameter Gal-immunoreactive fibers. These large-diameter Gal-immunoreactive fibers did not co-contain dopamine beta-hydroxylase (DBH). In contrast, virtually every small-caliber Gal-immunoreactive fiber colocalized with DBH. To determine whether Gal-immunoreactive fibers originated from LC, we combined immunohistochemical procedures with fluorescent tracing techniques. After retrograde tracer injections into several trigeminal relay nuclei, we observed that approximately 50% of the labeled LC neuronal population was immunoreactive for Gal. Our results suggest an extensive Gal-immunoreactive fiber innervation of the rodent trigeminal system, much of which may originate from LC neurons in the brainstem.
Assuntos
Galanina/análise , Proteínas do Tecido Nervoso/análise , Tálamo/química , Nervo Trigêmeo/química , Núcleos do Trigêmeo/química , Vias Aferentes/química , Vias Aferentes/ultraestrutura , Animais , Dopamina beta-Hidroxilase/análise , Feminino , Microscopia de Fluorescência , Ratos , Sinapses/químicaRESUMO
In previous work it has been shown that adult male, but not female, offspring of rats that have been injected with protamine zinc insulin (6 IU/kg) on days 15-20 of gestation, develop significant obesity beginning about 50 days of age. This obesity is accompanied by elevated medial hypothalamic extracellular norepinephrine levels. To examine whether the expression of obesity in male offspring is mediated by perinatal testosterone levels, male offspring of insulin-treated or control dams were either castrated or received sham surgery on postnatal day 1. Castrated male offspring of insulin-treated dams did not become obese like their gonadally intact male littermates. This suggests that perinatal testosterone levels may interact with developmental processes mediating the obesity in male offspring of insulin-treated dams. A second question addressed was whether the elevated hypothalamic extracellular norepinephrine levels observed in our earlier work are evident as morphological changes in norepinephrine-containing systems in the medial hypothalamus and locus coeruleus. We found a significant enhancement of dopamine-beta-hydroxylase immunoreactivity in fibers innervating the paraventricular nucleus of the hypothalamus in 121-day-old, gonadally intact male offspring of insulin-injected dams. This suggests that the impact of maternal insulin injections on offspring obesity may be mediated through its organizing action on feeding-related fibers in the paraventricular nucleus.
Assuntos
Peso Corporal/fisiologia , Hipotálamo/embriologia , Insulina/sangue , Norepinefrina/fisiologia , Efeitos Tardios da Exposição Pré-Natal , Animais , Dopamina beta-Hidroxilase/análise , Comportamento Alimentar/fisiologia , Feminino , Gônadas/fisiologia , Gônadas/cirurgia , Hipotálamo/química , Hipotálamo/enzimologia , Imuno-Histoquímica , Masculino , Vias Neurais , Obesidade/embriologia , Obesidade/fisiopatologia , Orquiectomia , Gravidez , Prosencéfalo/química , Prosencéfalo/embriologia , Prosencéfalo/enzimologia , Ratos , Ratos Sprague-DawleyRESUMO
Immunocytochemical staining for the presence of catecholamine synthesizing enzymes, tyrosine hydroxylase and dopamine beta-hydroxylase, was used to characterize the regional distribution of catecholaminergic neurons in the hypothalamus and adjacent areas of domestic cattle, Bos taurus. In steers, heifers and cows, tyrosine hydroxylase-immunoreactive perikarya was located throughout periventricular regions of the third cerebral ventricle, in both anterior and retrochiasmatic divisions of the supraoptic nucleus, suprachiasmatic nucleus, and ventral and dorsolateral regions of the paraventricular nucleus, dorsal hypothalamus, ventrolateral aspects of the arcuate nucleus, along the ventral hypothalamic surface between the median eminence and optic tract, and in the posterior hypothalamus. Immunostained perikarya ranged from small (10-20 microns, parvicellular) to large (30-50 microns, magnocellular) and were of multiple shapes: round, triangular, fusiform or multipolar, often with 2-5 processes of branched arborization. There were no dopamine-beta-hydroxylase immunoreactive perikarya observed within the hypothalamus and adjacent structures. However, both tyrosine hydroxylase and dopamine-beta-hydroxylase immunoreactive fibers and punctate varicosities were observed throughout regions of tyrosine hydroxylase immunoreactivity perikarya. Generally, the location and pattern of hypothalamic tyrosine hydroxylase immunoreactivity and dopamine-beta-hydroxylase immunoreactive were similar to those reported for most other large brain mammalian species, however, there were several differences with commonly used small laboratory animals. These included intense tyrosine hydroxylase immunoreactivity of perikarya within the retrochiasmatic division of the supraoptic nucleus (ventral A15 region), the absence of tyrosine hydroxylase immunoreactive perikarya below the anterior commissure or within the bed nucleus of stria terminalis (absence of the dorsal A15 region), an abundance of tyrosine hydroxylase immunoreactive perikarya within the ependymal layer of the median eminence, heavy innervation of the arcuate nucleus with dopamine-beta-hydroxylase immunoreactive fibers and varicosities, and the paucity of dopamine-beta-hydroxylase immunoreactive throughout the median eminence.
Assuntos
Catecolaminas/biossíntese , Dopamina beta-Hidroxilase/análise , Hipotálamo/enzimologia , Tirosina 3-Mono-Oxigenase/análise , Animais , Avidina , Bovinos , Ventrículos Cerebrais/anatomia & histologia , Ventrículos Cerebrais/enzimologia , Feminino , Hipotálamo/anatomia & histologia , Técnicas Imunoenzimáticas , Imuno-Histoquímica , MasculinoRESUMO
The effects of repeated cocaine administration on cochlear function were evaluated by measuring amplitude-intensity and latency-intensity functions of the whole-nerve action potential of the auditory nerve. Whole-nerve action potential input/output functions obtained using tone-pips of 0.5, 1, 2, 4 and 8 kHz in a group of cocaine-treated subjects were compared with those obtained in saline-treated animals. All measurements were made 24 h after the last treatment. Amplitudes of whole-nerve action potentials were enhanced in the cocaine-treated animals compared to the control group. No statistically significant differences in latency-intensity functions were seen after cocaine treatment. The effect of chronic cocaine exposure also was examined on catecholamine innervation in the cochlea using immunohistochemical techniques. The density of adrenergic innervation was reduced in the cocaine-treated animals.
Assuntos
Cocaína/farmacologia , Cóclea/efeitos dos fármacos , Estimulação Acústica , Fibras Adrenérgicas/efeitos dos fármacos , Animais , Chinchila , Cocaína/administração & dosagem , Cóclea/irrigação sanguínea , Cóclea/química , Cóclea/metabolismo , Cóclea/fisiologia , Dopamina beta-Hidroxilase/análise , Esquema de Medicação , Imuno-Histoquímica , Ketamina/farmacologia , Norepinefrina/metabolismo , Órgão Espiral/efeitos dos fármacos , Tirosina 3-Mono-Oxigenase/análiseRESUMO
A detailed study comparing the distribution of D2 receptors and tyrosine hydroxylase-immunoreactive fibers in the hippocampus and parahippocampal cortices of the rat, cat, and human was conducted. The distribution of [125I]epidepride binding to D2 receptors along the transverse and longitudinal axes of the hippocampus and parahippocampus differed among the species. In rat hippocampus, the number of sites was highest in septal portions of lacunosum-moleculare of CA1 and stratum moleculare of the subiculum. Virtually no binding to D2 receptors existed in the temporal hippocampus. For the cat hippocampus, the highest binding existed in the inner one-third of the molecular layer of the dentate gyrus (DG). There were also significant numbers of D2 receptors in strata radiatum and oriens of the CA subfields, with almost undetectable levels in lacunosum moleculare and subiculum. The number of sites was higher in the septal than temporal hippocampus. In the human hippocampus, highest binding was observed in the molecular layer of DG and the subiculum, with lower levels in strata oriens and lacunosum-moleculare of CA3, and very low binding in CA1. The histochemical demonstration of the pattern of mossy fibers revealed an organization complementary to that of D2 receptors in cat and human. In none of the species was there significant expression of D2 receptors in the entorhinal cortex, except in the caudal extreme of this region in the rat. In that region a trilaminar pattern was exhibited that continued into the perirhinal cortex. A trilaminar pattern of D2 receptor expression was observed in the perirhinal cortex of all species, with the highest values in the external and deep laminae and low expression in the middle laminae. The organization of dopamine fibers was assessed by comparing the distribution of tyrosine hydroxylase-positive and dopamine beta-hydroxylase-immunoreactive fibers in these same regions. It revealed consistent mismatches between the pattern of D2 receptor expression and dopaminergic innervation in all three species. The implications for this mismatch are discussed. It is hypothesized that the distribution of D2 receptors, and not of dopamine fibers, determines what neural systems dopamine influences in the hippocampal complex.
Assuntos
Hipocampo/química , Receptores de Dopamina D2/análise , Tirosina 3-Mono-Oxigenase/análise , Idoso , Idoso de 80 Anos ou mais , Animais , Autorradiografia , Benzamidas/metabolismo , Gatos , Dopamina beta-Hidroxilase/análise , Córtex Entorrinal/química , Feminino , Humanos , Imuno-Histoquímica , Radioisótopos do Iodo , Masculino , Fibras Nervosas/metabolismo , Pirrolidinas/metabolismo , Ratos , Receptores de Dopamina D2/metabolismoRESUMO
Transneuronal tracing techniques were used in order to identify putative spinal interneurons and brainstem sites involved in the control of penile function. Pseudorabies virus was injected into the corpus cavernosus tissue of the penis in rats. After a four day survival period, rats were perfused with fixative and virus-labelled neurons were identified by immunohistochemistry. Postganglionic neurons were retrogradely labelled in the major pelvic ganglia. In the spinal cord, sympathetic and parasympathetic preganglionic neurons were labelled transneuronally. Presumptive interneurons were also labelled in the lower thoracic and lumbosacral spinal cord in locations consistent with what is currently known about such interneurons. In the brainstem, transneuronally labelled neurons were found in the medulla, pons and hypothalamus. Regions consistently labelled included the nucleus paragigantocellularis, parapyramidal reticular formation of the medulla, raphe pallidus, raphe magnus, A5 noradrenergic cell group, Barrington's nucleus and the paraventricular nucleus of the hypothalamus. This study confirmed previous studies from our lab and others concerning the preganglionic and postganglionic neurons innervating the penis. The number, morphology and location of these neurons were consistent with labelling seen following injection of conventional tracers into the penis. The brainstem nuclei labelled in this study were also consistent with what is currently known about the brainstem control of penile function. The labelling appeared to be highly specific, in that descending systems involved in other functions were not labelled. These results provide further evidence that the pseudorabies virus transneuronal tracing technique is a valuable method for identifying neural circuits mediating specific functions.
Assuntos
Fibras Autônomas Pós-Ganglionares/ultraestrutura , Fibras Autônomas Pré-Ganglionares/ultraestrutura , Transporte Axonal , Mapeamento Encefálico , Sistema Nervoso Central/anatomia & histologia , Dopamina beta-Hidroxilase/análise , Herpesvirus Suídeo 1 , Proteínas do Tecido Nervoso/análise , Pênis/inervação , Serotonina/análise , Vias Aferentes/ultraestrutura , Animais , Fibras Autônomas Pós-Ganglionares/química , Fibras Autônomas Pós-Ganglionares/microbiologia , Fibras Autônomas Pré-Ganglionares/química , Fibras Autônomas Pré-Ganglionares/microbiologia , Contagem de Células , Sistema Nervoso Central/química , Sistema Nervoso Central/microbiologia , Sistema Nervoso Central/fisiologia , Ejaculação/fisiologia , Gânglios Parassimpáticos/química , Gânglios Parassimpáticos/microbiologia , Gânglios Parassimpáticos/ultraestrutura , Herpesvirus Suídeo 1/isolamento & purificação , Hipotálamo/química , Hipotálamo/microbiologia , Hipotálamo/fisiologia , Hipotálamo/ultraestrutura , Interneurônios/química , Interneurônios/microbiologia , Interneurônios/ultraestrutura , Masculino , Bulbo/química , Bulbo/microbiologia , Bulbo/fisiologia , Bulbo/ultraestrutura , Ereção Peniana/fisiologia , Pênis/fisiologia , Ponte/química , Ponte/microbiologia , Ponte/fisiologia , Ponte/ultraestrutura , Núcleos da Rafe/química , Núcleos da Rafe/microbiologia , Núcleos da Rafe/fisiologia , Núcleos da Rafe/ultraestrutura , Ratos , Ratos Sprague-Dawley/anatomia & histologia , Medula Espinal/química , Medula Espinal/microbiologia , Medula Espinal/fisiologia , Medula Espinal/ultraestruturaRESUMO
Immunodetection of catecholamine biosynthetic enzymes is frequently used for the visualization of central nervous catecholaminergic systems. Because of the method's limited specificity for the different catecholamines, interpretation of the results often presents difficulties. To determine criteria for the identification of dopaminergic, noradrenergic, and adrenergic afferents to the rat amygdaloid complex, comparative immunolabelling for tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH), and phenylethanolamine-N-methyl-transferase (PNMT) was carried out using single- and double-labelling for fluorescence, light- and electron microscopy. The observations were complemented by findings in brainstem and hypothalamic areas. The results indicated that TH-labelling detected preferentially dopaminergic afferents in the lateral central and intercalated amygdaloid nuclei. DBH-labelling detected noradrenergic axons in nuclei lacking PNMT-immunoreactive fibres, and PNMT was a marker for adrenergic axons in the entire complex. For nuclei with combined dense dopaminergic, noradrenergic and/or adrenergic innervation, morphological and immunolabelling characteristics were described which, to a certain extent, enabled identification of the different afferents in anti-TH or anti-DBH-preparations. Using a monoclonal TH-antiserum, noradrenergic and adrenergic axons displayed weaker immunoreactivity than dopaminergic ones, and possessed characteristic morphological features. TH-immunoreactivity in noradrenergic axons differed depending on their origin, and showed intra-axonal compartmentalization. The present study provides a basis for the use of the detection of biosynthetic enzymes in future investigations into the ultrastructure and connectivity of the catecholaminergic amygdala innervation.
Assuntos
Tonsila do Cerebelo/química , Dopamina beta-Hidroxilase/análise , Imuno-Histoquímica , Feniletanolamina N-Metiltransferase/análise , Tirosina 3-Mono-Oxigenase/análise , Tonsila do Cerebelo/anatomia & histologia , Animais , Axônios/química , Hipotálamo/química , Masculino , Microscopia Eletrônica , Ratos , Ratos WistarRESUMO
The distribution of noradrenergic processes within the hypothalamus of rhesus monkeys (Macaca mulatta) was examined by immunohistochemistry with an antibody against dopamine-beta-hydroxylase. The results revealed that the pattern of dopamine-beta-hydroxylase immunoreactivity varied systematically throughout the rhesus monkey hypothalamus. Extremely high densities of dopamine-beta-hydroxylase-immunoreactive processes were observed in the paraventricular and supraoptic nuclei, while relatively lower levels were found in the arcuate and dorsomedial nuclei and in the medial preoptic, perifornical, and suprachiasmatic areas. Moderate levels of dopamine-beta-hydroxylase immunoreactivity were found throughout the lateral hypothalamic area and in the internal lamina of the median eminence. Very few immunoreactive processes were found in the ventromedial nucleus or in the mammillary complex. Other midline diencephalic structures were found to have high densities of dopamine-beta-hydroxylase immunoreactivity, including the paraventricular nucleus of the thalamus and a discrete subregion of nucleus reuniens, the magnocellular subfascicular nucleus. A moderate density of dopamine-beta-hydroxylase immunoreactive processes were found in the rhomboid nucleus and zona incerta whereas little dopamine-beta-hydroxylase immunoreactivity was found in the fields of Forel, nucleus reuniens, or subthalamic nucleus. The differential distribution of dopamine-beta-hydroxylase-immunoreactive processes may reflect a potential role of norepinephrine as a regulator of a variety of functions associated with the nuclei that are most heavily innervated, e.g., neuroendocrine release from the paraventricular and supraoptic nuclei, and gonadotropin release from the medial preoptic area and mediobasal hypothalamus. Additionally, quantitative analysis of dopamine-beta-hydroxylase-immunoreactive varicosities was performed on a laser scanning microscope in both magnocellular and parvicellular regions of the paraventricular nucleus of the hypothalamus. The methodology employed in this study allowed for the high resolution of immunoreactive profiles through the volume of tissue being analyzed, and was more accurate than conventional light microscopy in terms of varicosity quantification. Quantitatively, a significant difference in the density of dopamine-beta-hydroxylase-immunoreactive varicosities was found between magnocellular and parvicellular regions, suggesting that parvicellular neurons received a denser noradrenergic input. These differential patterns may reflect an important functional role for norepinephrine in the regulation of anterior pituitary secretion through the hypothalamic-pituitary-adrenal stress axis.
Assuntos
Dopamina beta-Hidroxilase/análise , Hipotálamo/enzimologia , Macaca mulatta/metabolismo , Fibras Nervosas/química , Norepinefrina/fisiologia , Núcleo Hipotalâmico Paraventricular/irrigação sanguínea , Animais , Imuno-Histoquímica , Núcleo Hipotalâmico Paraventricular/enzimologia , Núcleos Talâmicos/enzimologia , Veias/químicaRESUMO
Using traditional as well as whole-mount immunohistochemistry, we described the location of tyrosine hydroxylase- and dopamine beta hydroxylase-positive cells and fibers in the brain of the lizard Anolis carolinensis. Major catecholaminergic cell groups were in the ependyma in certain ventricular regions, along the periventricular floor in the preoptic region, within the anterior hypothalamic and lateral hypothalamic areas, and in the mesencephalic tegmental region, locus coeruleus, nucleus of the solitary tract, vagal motor nucleus, and rhombencephalic reticular formation. Major catecholaminergic fibers, tracts and varicosities included tuberohypophysial, mesolimbic, nigrostriatal, isthmocortical, medullohypothalamic, and coeruleospinal systems. Although the catecholaminergic systems in A. carolinensis are similar to those in the brains of other lizards studied, there are a few species differences. Our information about A. carolinensis will be used to help localize the hypothalamic asymmetry in catecholamine metabolism previously described in this lizard.
Assuntos
Encéfalo/citologia , Dopamina/fisiologia , Lagartos/anatomia & histologia , Norepinefrina/fisiologia , Animais , Biomarcadores , Mapeamento Encefálico , Dopamina beta-Hidroxilase/análise , Vias Eferentes/anatomia & histologia , Epêndima/citologia , Estro , Feminino , Lateralidade Funcional , Hipotálamo/citologia , Hipotálamo/fisiologia , Imuno-Histoquímica/métodos , Proteínas do Tecido Nervoso/análise , Ovário/inervação , Ovário/fisiologia , Especificidade da Espécie , Medula Espinal/anatomia & histologia , Medula Espinal/fisiologia , Tirosina 3-Mono-Oxigenase/análiseRESUMO
The presence of 'tyrosine hydroxylase (TH)-only'-immunoreactive (IR) neurons is reported for the first time in the human hypothalamic region and basa ganglia, using immunohistochemical technique. 'TH-only'-IR neurons were demonstrated in the ventrolateral part of the arcuate nucleus and medial segment of globus pallidus. These neurons lacked aromatic L-amino acid decarboxylase (AADC)-, dopamine-beta-hydroxylase (DBH)-, and dopamine (DA)-immunoreactivity. The above results suggest that these 'TH-only'-IR neurons are not dopaminergic, and possibly contain L-DOPA as an end-product.
Assuntos
Núcleo Arqueado do Hipotálamo/metabolismo , Globo Pálido/metabolismo , Levodopa/metabolismo , Neurônios/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismo , Descarboxilases de Aminoácido-L-Aromático/análise , Gânglios da Base/metabolismo , Dopamina/análise , Dopamina beta-Hidroxilase/análise , Humanos , Hipotálamo/metabolismo , Recém-Nascido , Masculino , Tirosina 3-Mono-Oxigenase/análiseRESUMO
The noradrenergic system interacts with steroid hormones to influence female sexual behavior and gonadotropin release in rodents. Using a double label immunocytochemical procedure for estrogen receptors and dopamine-beta-hydroxylase, we investigated the anatomical relationships between noradrenergic neurons and estrogen receptor-immunoreactive (ER-IR) cells in the brain of ovariectomized female guinea pigs. Dopamine-beta-hydroxylase-immunoreactive (DBH-IR) varicosities were closely associated with ER-IR cells throughout the hypothalamus and preoptic area. This anatomical relationship was observed with almost 80% of the ER-IR cells in the ventrolateral hypothalamus (VLH), an area involved in the regulation of female sexual behavior in guinea pigs. Furthermore, the presence of closely associated DBH-IR varicosities was related to staining intensity of ER-IR neurons. In the rostral VLH, ER-IR neurons with closely associated DBH-IR varicosities stained more darkly than ER-IR neurons lacking this association, suggesting noradrenergic regulation of basal levels of cellular estrogen receptors. These findings provide neuroanatomical evidence suggestive of noradrenergic regulation of estrogen receptor levels in the hypothalamus of female guinea pigs.
Assuntos
Dopamina beta-Hidroxilase/análise , Hipotálamo Médio/química , Hipotálamo/química , Norepinefrina/fisiologia , Área Pré-Óptica/química , Receptores de Estrogênio/análise , Animais , Feminino , Cobaias , Hipotálamo/irrigação sanguínea , Técnicas Imunoenzimáticas , Neurônios/química , Perfusão , Área Pré-Óptica/irrigação sanguínea , Varizes/metabolismoRESUMO
To determine the central neural pathway which carries splanchnic osmosensory information to vasopressin (AVP) neurons in the hypothalamus, bilateral electrolytic lesions were placed in the ascending catecholaminergic fiber bundle, the locus coeruleus (LC), the locus subcoeruleus (subLC), the lateral parabrachial nucleus (LPB), the caudal periaqueductal gray (PAG) and the median preoptic nucleus (MPO). Six and seven days later, plasma AVP levels, plasma osmolality, mean arterial pressure and heart rate were measured following gastric infusion of hypertonic (598 mosm/kg; 2 ml/4 min) or isotonic (290 mosm/kg) saline in conscious rats with indwelling tail artery catheters and nasogastric tubes. The most effective pontine lesions, which were located in the ventral locus subcoeruleus (vsubLC) approximately 1.0 mm below the LC, decreased the AVP response to hypertonic gastric infusion by 59.7% (P less than 0.05) as compared to sham-lesioned controls. In addition, unilateral vsubLC lesions dramatically reduced the catecholamine innervation of the ipsilateral paraventricular nucleus (PVN), as qualitatively determined with dopamine beta-hydroxylase immunocytochemistry, suggesting that a pathway ascending with catecholaminergic fibers was disrupted. Lesions of the MPO were also very effective, decreasing the AVP response to hypertonic saline infusion by 60.3% (P less than 0.05), suggesting that the MPO is an integral relay center in this pathway. On the other hand, LC, LPB and PAG lesions were ineffective. Systemic plasma osmolality or cardiovascular factors did not mediate the AVP response. These results demonstrate, for the first time, that splanchnic osmotic information is transmitted to the hypothalamus via pathways within the ascending catecholaminergic fiber bundles, the MPO is a relay center where peripheral and central osmotic information may be integrated, and the LC, LPB, and PAG are not part of the splanchnic osmotic pathway.
Assuntos
Vias Aferentes/fisiologia , Arginina Vasopressina/fisiologia , Encéfalo/fisiologia , Hipotálamo/fisiologia , Mecanorreceptores/fisiologia , Neurônios/fisiologia , Solução Salina Hipertônica/farmacologia , Cloreto de Sódio/farmacologia , Circulação Esplâncnica/fisiologia , Nervos Esplâncnicos/fisiologia , Equilíbrio Hidroeletrolítico , Animais , Arginina Vasopressina/sangue , Dopamina beta-Hidroxilase/análise , Hipotálamo/efeitos dos fármacos , Masculino , Neurônios/efeitos dos fármacos , Ratos , Ratos EndogâmicosRESUMO
Bone metabolism may be influenced by the innervation of skeletal tissues. Neuropeptides such as vasoactive intestinal peptide, from sympathetic nerves, and calcitonin gene-related peptide, from sensory nerves, have been implicated as local modulators of bone metabolism. The effect of neonatal sympathectomy and of capsaicin-induced sensory denervation in rats was studied on the following: (i) the radial bone growth and apposition rate in tibiae (normal growth and modeling) and (ii) the percentage of periosteal surface of the mandible occupied by osteoclasts during induced remodeling. Neonate rats were treated with guanethidine, capsaicin, or appropriate vehicle. At seven weeks, maxillary molars were removed to induce remodeling on the buccal surface of the mandible. Animals were killed four days after surgery. Cross-sectional cortical area, medullary area, and periosteal apposition rate were measured by histomorphometry in ground sections of tibiae. The percentage of periosteal surface at the remodeling site occupied by osteoclasts (stained for acid phosphatase) was measured in frozen, undecalcified sections. There was no significant difference in cortical or medullary area or periosteal apposition rate in tibiae between each drug treatment and its control. However, the mandibular bone surface occupied by osteoclasts was increased 45.5% (P less than or equal to 0.005) in animals treated neonatally with guanethidine compared to controls. In contrast, the mandibular surface occupied by osteoclasts was decreased 21.2% (P less than or equal to 0.04) in animals treated neonatally with capsaicin compared to controls. The alteration of bone remodeling (osteoclast surface) by both treatments indicates that sensory and sympathetic nerves play a role in focal metabolism of bone.
Assuntos
Animais Recém-Nascidos/fisiologia , Remodelação Óssea/efeitos dos fármacos , Peptídeo Relacionado com Gene de Calcitonina/fisiologia , Capsaicina , Simpatectomia Química , Peptídeo Intestinal Vasoativo/fisiologia , Animais , Remodelação Óssea/fisiologia , Peptídeo Relacionado com Gene de Calcitonina/análise , Dopamina beta-Hidroxilase/análise , Guanetidina , Masculino , Mandíbula/inervação , Fibras Nervosas/efeitos dos fármacos , Fibras Nervosas/fisiologia , Fibras Nervosas Mielinizadas/efeitos dos fármacos , Fibras Nervosas Mielinizadas/fisiologia , Neurônios Aferentes/efeitos dos fármacos , Neurônios Aferentes/fisiologia , Osteoclastos/patologia , Ratos , Ratos Endogâmicos , Substância P/análise , Tíbia/inervação , Tíbia/patologia , Peptídeo Intestinal Vasoativo/análiseRESUMO
There is considerable evidence from biochemical studies that the transmitter-depleting action of drugs and neurotoxins which act upon central noradrenergic (NA) axon terminals is not uniform in different brain regions. Among NA axons, those originating in the locus coeruleus (LC) have been proposed to be most susceptible to the action of NA neurotoxins such as N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4). The studies described here were conducted to determine whether this differential susceptibility to DSP-4 reflects a pharmacological heterogeneity between different populations of NA axons. To determine whether DSP-4 acts selectively upon LC axons, we have characterized the effects of this drug on NA axons in different brain regions, by using noradrenaline and dopamine-beta-hydroxylase (D beta H) immunohistochemistry. Following systemic administration of DSP-4, there was an almost complete loss of noradrenaline and D beta H staining in brain regions innervated by LC axons. No effects of the drug treatment were detected in brain regions innervated primarily by non-coerulean NA axons. These results demonstrate that both the transmitter-depleting and the neurodegenerative action of DSP-4 are restricted to NA axons originating in the LC. To explore the basis for this selectivity, noradrenaline uptake studies were conducted using synaptosomes from brain regions in which NA axons differ in their response to DSP-4. The results reveal a significant difference in the affinity of DSP-4 for the noradrenaline uptake carrier in cortical and hypothalamic synaptosomes. This finding is compatible with the hypothesis that the noradrenaline uptake carrier is pharmacologically distinct in LC and non-coerulean NA axons. This heterogeneity in noradrenaline uptake raises the question whether other drugs may also have differential actions on LC and non-coerulean NA neurons.
Assuntos
Adrenérgicos/farmacologia , Fibras Adrenérgicas/efeitos dos fármacos , Benzilaminas/farmacologia , Locus Cerúleo/efeitos dos fármacos , Inibidores da Captação de Neurotransmissores/farmacologia , Norepinefrina/fisiologia , Adrenérgicos/toxicidade , Axônios/efeitos dos fármacos , Benzilaminas/toxicidade , Córtex Cerebral/citologia , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Dopamina beta-Hidroxilase/análise , Resistência a Medicamentos , Hipotálamo/citologia , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Degeneração Neural/efeitos dos fármacos , Inibidores da Captação de Neurotransmissores/toxicidade , Oxidopamina/toxicidade , Sinaptossomos/efeitos dos fármacos , Sinaptossomos/metabolismoRESUMO
Antisera directed against human dopamine-beta-hydroxylase and against serotonin were used to characterize the noradrenergic (NA) and serotoninergic (5-HT) innervation of several cortical and subcortical visual areas in squirrel monkey (Saimiri sciureus) and cynomolgus monkey (Macaca fascicularis). Few species differences were observed for either monoamine. Cortical areas 17 and 18, as well as visual areas in the temporal and parietal lobe were found to exhibit regional specialization of both 5-HT and NA innervation. Precisely at the border between areas 17 and 18, the laminar innervation patterns and density characteristic of NA fibers in area 17 (Morrison et al., '82a; Kosofsky et al., '84) shift so that layer IV of area 18 contains more fibers than layer IV of area 17, and the overall density of fibers in area 18 is higher. For 5-HT, the highly laminated patterns characteristic of area 17 (Morrison et al., '82a; Kosofsky et al., '84) also observe this cytoarchitectonic boundary. Fibers in area 18 are more evenly distributed across laminae, and the overall density of fibers decreases. The visual region of the inferotemporal cortex was found to be very lightly innervated by NA fibers and very densely innervated by 5-HT fibers. Area 7 of the parietal lobule was more densely innervated by NA fibers, and less densely innervated by 5-HT fibers, than any other visual cortical region examined. The visual thalamic nuclei exhibited even greater regional differences in the density of NA innervation. The lateral geniculate nucleus was found to be virtually devoid of NA fibers, while the pulvinar-lateral posterior complex was densely innervated. The density of 5-HT fibers was more uniform across thalamic visual nuclei. The lateral geniculate, pulvinar, and lateral posterior nuclei all exhibit a moderate to high density of immunoreactive fibers. In the mesencephalon, the superficial layers of the superior colliculus were found to be densely innervated by NA fibers, whereas 5-HT fibers were most dense in the intermediate layers. These patterns of innervation indicate that, in these primate species, functionally related visual regions share common and distinguishable densities of NA innervation. Specifically, tecto-pulvinar-juxtastriate structures are more densely innervated than geniculo-striate and inferotemporal structures. These relationships suggest that, within the visual system, NA fibers preferentially innervate the regions involved in spatial analysis and visuomotor response rather than those involved in feature extraction and pattern analysis.(ABSTRACT TRUNCATED AT 400 WORDS)