RESUMO
The posttranslational processing of prodynorphin (Pro-Dyn) is not well understood. The rat anterior pituitary is an interesting tissue which merits examination to address this issue since it is known that Dyn immunoreactivity is stored as high mol wt (HMW) intermediates and not as free products such as dynorphin-A-(1-17) (Dyn-A17) or dynorphin-B-(1-13) (Dyn-B). The aim of our study is to characterize the Pro-Dyn products in the rat anterior pituitary quantitatively as well as qualitatively by keeping a close account of each of the possible domains that are known to compose the protein structure. This was achieved by a convergence of tools: designing RIA with antibodies to each of these domains, including antibodies to Dyn-A17, Dyn-B, alpha-neo-endorphin, bridge peptide, and Pro-Dyn carboxyl-terminal peptide (C-peptide), and using these antisera with gel filtration chromatography, reverse phase HPLC, immunoaffinity, and immunoprecipitation techniques. Our data indicate the presence of at least six distinct molecules which are classified as HMW intermediates (greater than 3.5K). By gel filtration chromatography they have apparent mol wt of 16,000, 10,000, 8,000, 6,000, 4,000, and 3,500, respectively. Each of these structures is characterized by multiple immunoreactivities to account for the observed mass. Based on the relative content of each structure we present a scheme for the posttranslational processing pathway of Pro-Dyn in the rat anterior pituitary. We also analyze other tissues, spinal cord and hypothalamus, for their content of Pro-Dyn HMW intermediates. Our results indicate that these tissues store Pro-Dyn HMW molecules of similar sizes and immunoreactive properties, suggesting that Pro-Dyn may be processed in a similar manner, at least in the initial phases, across tissues.
Assuntos
Encefalinas/metabolismo , Adeno-Hipófise/metabolismo , Precursores de Proteínas/metabolismo , Processamento de Proteína Pós-Traducional , Animais , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Hipotálamo/análise , Técnicas Imunológicas , Masculino , Peso Molecular , Adeno-Hipófise/análise , Testes de Precipitina , Radioimunoensaio , Ratos , Ratos Endogâmicos , Medula Espinal/análise , Extratos de Tecidos/análiseRESUMO
With the techniques of specific radioimmunoassay and gel filtration it was found that CGRP was distributed in various tissues of normotensive (WKY) and spontaneously hypertensive rats (SHR) with the highest concentration in the lumbar spinal cord (1197 +/- 94.8 pg/mg tissue) and the lowest in the auricle (15.0 +/- 2.1 pg/mg tissue). In comparison with WKY, CGRP concentration in the plasma was decreased and in the abdominal aorta and hypothalamus was increased in SHR. Gel filtration revealed only one major CGRP molecular form in the tissues. In addition, CGRP reduced the mean arterial pressure (MAP) in SHR in a dose-dependent manner. These data suggest that CGRP may play an important role in the pathogenesis of hypertension and its possible therapy.
Assuntos
Calcitonina/análise , Neuropeptídeos/análise , Ratos Endogâmicos SHR/metabolismo , Ratos Endogâmicos/metabolismo , Animais , Aorta Abdominal/análise , Pressão Sanguínea/efeitos dos fármacos , Peptídeo Relacionado com Gene de Calcitonina , Frequência Cardíaca/efeitos dos fármacos , Hipotálamo/análise , Músculo Liso Vascular/análise , Miocárdio/análise , Neuropeptídeos/sangue , Neuropeptídeos/farmacologia , Especificidade de Órgãos , Radioimunoensaio , Ratos , Ratos Endogâmicos WKY/metabolismo , Valores de Referência , Medula Espinal/análiseRESUMO
The morphological research of the acupoint began from 1977. The scholars have studied on three main works as following. 1) The sensory organs in the acupoint: The area of acupoint contains free nerve ending, Vater-Pacini corpuclese and muscular spindle. The nerve fibers are I, II, III types, the chief is II. 2) The segmental distributions of afferent neuronal fibers of the acupoint: The evidence of the experimental research has revealed that the fibers of mainly and essential acupoints are definite to be afferent segments. The fibers of the upper and lower limbs acupoints projected into III-IX lamina of the posterior horn of the spinal cord. 3) The mutual intersect in the dorsal ganglia between the segmental distributions of the afferent neurons of viscera and that of acupoints. It is proposed that the mutual intersect between the segmental distributions of the afferent neurons of viscera and that of the acupoints is probable the morphological foundation for the theory of somato-viscera.
Assuntos
Pontos de Acupuntura , Pele/inervação , Humanos , Fibras Nervosas , Neurônios Aferentes/citologia , Células Receptoras Sensoriais/anatomia & histologia , Medula Espinal/análise , Vísceras/inervaçãoRESUMO
The influence of adjuvant-induced arthritis of the rat on central and peripheral peptide neuroanatomy was investigated by immunohistochemistry. The most striking feature of arthritic rats was the differential intensification of neuronal proenkephalin- and prodynorphin-related staining in dorsal horn. Changes were ipsilateral in monoarthritic and bilateral in polyarthritic rats as compared to controls. Opioid responsive neurons were target of substance P (SP) and calcitonin gene-related peptide (CGRP) fibers. Changes of SP and CGRP predominated in peripheral inflamed tissue and consisted of intensified immunostaining and an apparent sprouting of sensory fibers particularly around venules, in the epidermis and in areas infiltrated by immunocompetent cells. Opioid staining was absent from primary afferents but present in some immune cells of inflamed tissue. Endogenous antinociceptive opioids and pro-nociceptive/pro-inflammatory SP and CGRP may be crucial in the concerted response of the neuroimmune system to chronic inflammatory pain.
Assuntos
Artrite Experimental/metabolismo , Artrite/metabolismo , Gânglios Espinais/análise , Fibras Nervosas/análise , Neuropeptídeos/análise , Medula Espinal/análise , Animais , Artrite Experimental/patologia , Peptídeo Relacionado com Gene de Calcitonina , Encefalinas/análise , Imuno-Histoquímica , Precursores de Proteínas/análise , Ratos , Ratos Endogâmicos , Pele/inervação , Substância P/análiseRESUMO
Atrial natriuretic factor (ANF) was investigated in the rat spinal cord and hypothalamus using two radioimmunoassays. ANF was also quantified in both tissues of Spontaneously Hypertensive Rats and Dahl rats. Spinal cord and hypothalamus were found to be immunoreactive to proANF and its near-NH2- or near-COOH-terminal fragments. A major part of the extracted ANF was a COOH-terminal peptide smaller than or the same as ANF (Ser99-Tyr 126). SHR had higher hypothalamic and spinal cord ANF concentrations than Wistar Kyoto rats, while the Dahl salt-sensitive animals exhibited an increase in spinal cord ANF when compared with the Dahl salt-resistant group. The data suggest that spinal cord may produce ANF locally with processing similar to that in hypothalamus. Changes in ANF concentrations occurring during the course of hypertension remain to be further investigated.
Assuntos
Fator Natriurético Atrial/análise , Ratos Endogâmicos SHR/metabolismo , Ratos Endogâmicos/metabolismo , Medula Espinal/análise , Animais , Cromatografia Líquida de Alta Pressão , Hipotálamo/análise , Ratos , Ratos Endogâmicos WKY/metabolismo , Valores de ReferênciaRESUMO
Methods for making and using hollow microdialysis fibers suitable for recovering extracellular substances from discrete regions of the spinal cord are described. After placement of the fiber, artificial cerebrospinal fluid was pushed through it at a low (4-5 microliters/min) rate. The perfusate was collected and samples analyzed on a high performance liquid chromatograph with an electrochemical detector. Serotonin, 5-hydroxyindole acetic acid and norepinephrine were recovered and identified. Single unit extracellular recordings were made during the perfusion and collection; thus simultaneous observation of neurotransmitter release and modulation of single cell activity is now possible.
Assuntos
Serotonina/análise , Medula Espinal/análise , Animais , Gatos , Cromatografia Líquida de Alta Pressão/métodos , Diálise/métodos , Feminino , Haplorrinos , Ácido Hidroxi-Indolacético/análise , Masculino , Potenciais da Membrana , Microquímica , Medula Espinal/fisiologia , Tálamo/fisiologiaRESUMO
Thyrotropin releasing hormone (TRH) and serotonin (5-HT) are co-transmitters in spinal and medullary neurons. To study the functional significance of this relationship and the regulation of TRH receptors, we chronically administered TRH and analogs MK-771 and CG-3509 to rats at a dose which evoked behavioral abnormalities. TRH reduced specific binding at spinal (24%) and hypothalamic (31%) TRH receptors and decreased TRH stimulated motor behaviors, such as rearing and cage crossing (locomotion). 5-HT1 and 5-HT2 receptors were unaffected except for an 11% increase in specific binding at spinal 5-HT1 sites. 5-HT and NE concentrations measured by HPLC were not altered in brainstem or hippocampus. In contrast, spinal TRH receptor specific binding increased 11% in rats treated intracisternally with 5,7-dihydroxytryptamine, but the effect was not significant. In competition studies in vitro, MK-771, TRH, and CG-3509 had no activity at 5-HT2 receptors in neocortex, brainstem, or spinal cord, and little activity at 5-HT1 sites (IC 50s greater than 100 uM). A mixed competitive and non-competitive binding profile at 5-HT1 sites resulted in the presence of 100 uM TRH. Conversely, 5-HT agonists had minimal effect at TRH receptors in spinal cord. These data suggest reciprocal or independent regulation of 5-HT1 and TRH receptors in co-transmitter and non-cotransmitter regions, respectively, in response to chronic TRH administration.
Assuntos
Receptores de Neurotransmissores/efeitos dos fármacos , Receptores de Serotonina/efeitos dos fármacos , Hormônio Liberador de Tireotropina/análogos & derivados , Hormônio Liberador de Tireotropina/administração & dosagem , Animais , Comportamento Animal/efeitos dos fármacos , Ligação Competitiva , Aminas Biogênicas/análise , Tronco Encefálico/análise , Córtex Cerebral/análise , Cromatografia Líquida de Alta Pressão , Hipotálamo/análise , Técnicas In Vitro , Locomoção/efeitos dos fármacos , Masculino , Ratos , Ratos Endogâmicos , Receptores de Neurotransmissores/fisiologia , Receptores de Serotonina/fisiologia , Receptores do Hormônio Liberador da Tireotropina , Medula Espinal/análise , Tiazolidinas , Hormônio Liberador de Tireotropina/farmacologiaRESUMO
Induction of experimental allergic encephalomyelitis (EAE) in Lewis rats by injection of guinea pig (GP) spinal cord homogenate (SCH) plus adjuvant (SCH-CFA) can be inhibited by treatment with the iron chelating agent desferrioxamine (DFOM). Interestingly, induction of EAE with purified myelin basic protein (BP-CFA) is not inhibited with DFOM. This dichotomy does not appear to be due to any quantitative differences in the two inocula since minimal clinical EAE produced by threshold levels of BP is not inhibited with DFOM. Passive EAE is not inhibited irrespective of the type of encephalitogen used to sensitize the donors. This suggests that the inhibitory effect of DFOM is acting on the afferent limb of the immune response to SCH-CFA. Injection of BP-CFA and SCH-CFA into the same site, mixing BP with central nervous system (CNS) lipids, or incorporating BP into liposomes, all induce EAE which can be partially inhibited by treatment with DFOM. These results support the hypothesis that the close association of lipids with the encephalitogen (i.e. BP) in SCH required extensive lipid breakdown before adequate antigen presentation can occur, and it is at this level that DFOM exerts its inhibitory effect.
Assuntos
Desferroxamina/uso terapêutico , Encefalomielite Autoimune Experimental/prevenção & controle , Proteína Básica da Mielina/imunologia , Medula Espinal/análise , Animais , Encefalomielite Autoimune Experimental/etiologia , Feminino , Cobaias , Imunização Passiva , Lipossomos , Proteína Básica da Mielina/administração & dosagem , Proteína Básica da Mielina/toxicidade , Ratos , Ratos Endogâmicos Lew , Baço/transplante , Extratos de Tecidos/imunologia , Extratos de Tecidos/toxicidadeRESUMO
A proctolin (PROC)-like peptide was studied immunohistochemically in the hypothalamus, lower brainstem and spinal cord of the rat using an antiserum against PROC conjugated to thyroglobulin. Neuronal cell bodies containing PROC-like immunoreactivity (PROC-LI) were observed in the dorsomedial, paraventricular and supraoptic nuclei of the hypothalamus and in the nucleus raphe magnus, nucleus raphe pallidus, nucleus raphe obscurus and nucleus interfascicularis nervi hypoglossi in the medulla oblongata. Fibers containing PROC-LI were seen in the median eminence and in other hypothalamic nuclei, and in the lower brainstem in cranial motor nuclei including the dorsal motor nucleus of the vagus nerve, the motor trigeminal nucleus, the facial nucleus and nucleus ambiguous, and in lower numbers in the nucleus of the solitary tract and locus coeruleus. Fibers containing PROC-LI were also located in the spinal cord, in the intermediolateral cell column at thoracic levels and in the ventral horns at all levels of the spinal cord. After transection of the spinal cord, all PROC-immunoreactive fibers below the lesion disappeared. Following injection of Fast blue into the thoracic spinal cord, retrogradely labeled cells in the nuclei raphe pallidus, obscurus and magnus and nucleus interfasciculari nervi hypoglossi were seen to contain PROC-LI. PROC-LI had a similar distribution as thyrotropin-releasing hormone (TRH)-LI in the above-mentioned areas and coexistence of TRH-LI and PROC-LI was shown in cell bodies in the hypothalamus and medulla oblongata. PROC-LI could also be shown to coexist with 5-hydroxytryptamine (5-HT)-LI in neuronal cell bodies in the lower brainstem. The results demonstrate the occurrence of a PROC-like peptide in the mammalian nervous system, and these neurons seem to be at least largely identical to previously described TRH systems. A possible involvement of the PROC-like peptide in spinal motor control is discussed in relation to the well-established role of PROC in control of motor behavior in insects and invertebrates.
Assuntos
Sistema Nervoso Central/análise , Neuropeptídeos , Oligopeptídeos/análise , Serotonina/análise , Hormônio Liberador de Tireotropina/análise , Animais , Imunofluorescência , Histocitoquímica , Hipotálamo/análise , Masculino , Bulbo/análise , Ratos , Ratos Endogâmicos , Medula Espinal/análiseRESUMO
Two human genes encoding precursors for two calcitonin gene-related peptides (CGRP) I (or alpha) and II (or beta) have been identified (Steenbergh, P. H., Höppener, J. W. M., Zandberg, J., Lips, C. J. M., and Jansz, H. S. (1985) FEBS Lett. 183, 403-407). The amino acid sequence of CGRP-I was obtained in medullary thyroid carcinoma extracts (Morris, H. R., Panico, M., Etienne, T., Tippins, J., Girgis, S. I., and MacIntyre, I. (1984) Nature 308, 746-748), but not in normal human tissues. The human CGRP-II peptide remained to be discovered. Here we have determined in the human spinal cord the amino acid composition and the partial amino acid sequence of the DNA-predicted CGRP-I and -II. The data indicate for the first time the existence of a second CGRP different from the known CGRP-I. CGRP-II has been identified in the central nervous system, pituitary, thyroid, and in medullary thyroid carcinoma as a major CGRP form together with CGRP-I.
Assuntos
Córtex Cerebelar/análise , Neuropeptídeos/isolamento & purificação , Hipófise/análise , Medula Espinal/análise , Tálamo/análise , Glândula Tireoide/análise , Neoplasias da Glândula Tireoide/análise , Sequência de Aminoácidos , Aminoácidos/análise , Peptídeo Relacionado com Gene de Calcitonina , Células Cultivadas , Humanos , Metástase Neoplásica , Especificidade de ÓrgãosAssuntos
Terapia por Acupuntura , Artrite Experimental/metabolismo , Artrite/metabolismo , Química Encefálica , Terapia por Estimulação Elétrica , Encefalina Leucina/análise , Medula Espinal/análise , Animais , Artrite Experimental/terapia , Feminino , Masculino , Hipófise/análise , Radioimunoensaio , Ratos , Ratos EndogâmicosRESUMO
This report presents evidence of changes in the concentration of monoamine neurotransmitters and their metabolites in homogenates of spinal cord and brain areas of male rats related to specific events of their mating behavior. Intact male rats were allowed to copulate with receptive females and decapitated immediately after either the first intromission or the first ejaculation. Non-mating control animals were exposed to other males, instead of females. The concentration of monoamines (norepinephrine, dopamine and serotonin) and some of their major metabolites (DOPAC and HIAA) in homogenates of discrete brain areas (parietal cortex, preoptic region, mediobasal hypothalamus) and lumbosacral spinal cord were measured by HPLC-ED. Results suggest that sexual arousal is associated with both increased dopaminergic activity in the preoptic region and inhibition of descending monoaminergic signals to the lumbosacral cord, whereas ejaculation is accompanied by increased activity of the serotonergic, as well as dopaminergic, innervation of the preoptic region. These findings give neurochemical support to notions of central monoamines involvement in sexual behavior suggested by previous pharmacological studies.
Assuntos
Química Encefálica , Neurotransmissores/análise , Comportamento Sexual Animal , Animais , Córtex Cerebral/análise , Córtex Cerebral/metabolismo , Hipotálamo/análise , Hipotálamo/metabolismo , Masculino , Neurotransmissores/metabolismo , Área Pré-Óptica/análise , Área Pré-Óptica/metabolismo , Ratos , Ratos Endogâmicos , Medula Espinal/análise , Medula Espinal/metabolismoRESUMO
Rats were given 75 mg/kg of 5,5-diphenylhydantoin (phenytoin) or vehicle 30 min prior to 75 mg/kg of 1,1,1-trichloro-bis(p-chlorophenyl)ethane (p,p'-DDT) (p.o.) or chlordecone (i.p.) and tremor was measured 12 h later. Rats were then killed, and regional brain levels of biogenic amines and their acid metabolites and amino acids were determined. Pretreatment with phenytoin significantly attenuated the tremor produced by p,p'-DDT but enhanced that produced by chlordecone. p,p'-DDT had significant effects on the levels of aspartate, glutamate, 5-hydroxyindoleacetic acid (5-HIAA), and 3-methoxy-4-hydroxyphenylglycol (MHPG), whereas chlordecone increased glycine, 5-HIAA, and MHPG levels. Pretreatment with phenytoin blocked p,p'-DDT-induced increases of aspartate in the brainstem and spinal cord, 5-HIAA in the hippocampus, and MHPG in the brainstem and hypothalamus. Phenytoin significantly enhanced chlordecone-induced increases of MHPG in the brainstem. These data indicate that organochlorine-induced increases in noradrenergic activity in the brainstem and spinal cord may be directly related to the tremorigenic effects of these chemicals.
Assuntos
Química Encefálica/efeitos dos fármacos , Clordecona/antagonistas & inibidores , DDT/antagonistas & inibidores , Inseticidas/antagonistas & inibidores , Fenitoína/farmacologia , Animais , Tronco Encefálico/análise , Núcleo Caudado/análise , Hipocampo/análise , Ácido Hidroxi-Indolacético/análise , Hipotálamo/análise , Masculino , Metoxi-Hidroxifenilglicol/análise , Norepinefrina/análise , Ratos , Ratos Endogâmicos F344 , Serotonina/análise , Medula Espinal/análise , Tremor/induzido quimicamenteRESUMO
We investigated the effects of chronic relapsing experimental allergic encephalomyelitis (first attack and first recovery) on uric acid (UR) concentrations in Lewis rat spinal cord. Perchloric extracts of 9 regions between spinal cord segments C3 and Co1 were injected into a reversed-phase high-performance liquid chromatographic system, and UR was quantified electrochemically. In all regions UR, which was lower than 0.5 micrograms/g wet tissue in rats injected with adjuvant (controls), increased to a variable extent during the attack (5-12 micrograms/g, maximum in T11-L2) and returned during recovery to a residual level of 1-2 micrograms/g. These findings can be partly attributed to blood-brain barrier damage.
Assuntos
Encefalomielite Autoimune Experimental/metabolismo , Medula Espinal/análise , Ácido Úrico/análise , Animais , Barreira Hematoencefálica , Encefalomielite Autoimune Experimental/fisiopatologia , Masculino , Ratos , Ratos Endogâmicos LewRESUMO
The time of appearance and tissue concentrations of substance P-like immunoreactivity (SP-LI) were studied in 53 human fetuses aged 8-21 weeks. Detectable amounts were present at 8 weeks of gestation in available fragments of spinal cord and intestine. Thereafter, the tissue concentrations were highest in spinal cord, intermediate in hypothalamus and lowest in digestive tract. Except for a significant increase in the intestinal wall, the concentrations did not vary from the 8-14 to the 15-21 week period. At chromatography, SP-LI in extracts of spinal cord and intestine was essentially eluted in the volume of the synthetic undecapeptide. Using the indirect immunofluorescence technique, the localization of SP-LI positive structures in the digestive tract was studied in 5 fetuses aged 12-18 weeks. Scarce cell bodies were observed in the myenteric plexus. Nerve fibers were recognized in the muscular layer, in the myenteric plexus and in connective tissue of pancreas. The present results demonstrate the early appearance of SP-LI positive structures both in central nervous system and in the enteric nervous system in the human fetus. In the age range tested, SP-LI concentrations were noticeably higher in spinal cord and hypothalamus than in the digestive tract.
Assuntos
Sistema Digestório/análise , Feto/análise , Hipotálamo/análise , Medula Espinal/análise , Substância P/análise , Sistema Digestório/embriologia , Desenvolvimento Embrionário e Fetal , Feto/fisiologia , Humanos , Hipotálamo/embriologia , Peptídeos/análise , Medula Espinal/embriologiaRESUMO
Most of the early studies on the immunohistochemical distribution of enkephalin pentapeptide-like immunoreactivity used antisera that stained both proenkephalin- and prodynorphin-containing neurons. The present study used the peroxidase-antiperoxidase method, thick Vibratome sections and antisera specific for the carboxyl termini of [Met]enkephalin, [Met]enkephalyl-Arg6-Phe7, [Met]enkephalyl-Arg6-Gly7-Leu8, and metorphamide and for BAM 22P in order to obtain a detailed description of the distribution of authentic proenkephalin-containing perikarya and nerve processes. The peroxidase-antiperoxidase reaction product was intensified by the selective deposition of silver crystals in order to display the morphology of proenkephalin-containing neurons with great fidelity. The results indicate that the magnocellular perikarya in the supraoptic and paraventricular nuclei contain prodynorphin rather than proenkephalin as had been suggested by earlier investigators. The coarse fibers in the internal zone of the median eminence and the granule cell-mossy fiber pathway in the hippocampus also contain prodynorphin rather than proenkephalin. The number of proenkephalin-containing perikarya and/or the density of proenkephalin-containing nerve terminals in several other areas of the brain, e.g. the substantia nigra, the central amygdaloid nucleus, the periaqueductal gray and the parabrachial nuclei, were overestimated by earlier investigators. The distribution of authentic proenkephalin-containing perikarya and nerve processes is, despite these errors, similar to the distribution of enkephalin pentapeptide-like immunoreactivity described by earlier investigators. Proenkephalin-containing perikarya were identified for the first time in the medial and lateral habenular nuclei of the adult rat. Antisera specific for [Met]enkephalin, [Met]enkephalyl-Arg6-Phe7, [Met]enkephalyl-Arg6-Gly7-Leu8 and BAM 22P stain perikarya and nerve terminals with a similar distribution. The metorphamide antiserum also stains the same perikarya and nerve terminals; however, it also stains magnocellular perikarya in the zona incerta and the lateral hypothalamus that are not stained by any of the other proenkephalin-specific antisera.
Assuntos
Encéfalo/citologia , Encefalinas/metabolismo , Neurônios/análise , Precursores de Proteínas/metabolismo , Animais , Encefalina Metionina/análogos & derivados , Encefalina Metionina/análise , Hipotálamo/citologia , Técnicas Imunoenzimáticas , Mesencéfalo/citologia , Neuro-Hipófise/citologia , Precursores de Proteínas/análise , Ratos , Ratos Endogâmicos , Medula Espinal/análise , Medula Espinal/citologia , Telencéfalo/citologia , Tálamo/citologiaAssuntos
Artrite/fisiopatologia , Endorfinas/fisiologia , (trans)-Isômero de 3,4-dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclo-hexil)-benzenoacetamida , Animais , Artrite Experimental/fisiopatologia , Benzomorfanos/farmacologia , Dinorfinas/sangue , Endorfinas/sangue , Encefalina Leucina/análogos & derivados , Encefalina Leucina/farmacologia , Encefalina Metionina/sangue , Morfina/farmacologia , Naloxona/farmacologia , Nociceptores/fisiologia , Dor/fisiopatologia , Adeno-Hipófise/análise , Precursores de Proteínas/sangue , Pirrolidinas/farmacologia , Ratos , Receptores Opioides/fisiologia , Limiar Sensorial , Medula Espinal/análise , Tálamo/análise , beta-EndorfinaRESUMO
Over the past decade our understanding of the localization of central cholinergic neurons has greatly increased. Interest in these systems has also intensified due to the involvement of cholinergic mechanisms in Alzheimer's disease. The distribution of central cholinergic neurons is reviewed, focusing on recent work in experimental animals. The pharmacohistochemical procedure for acetylcholinesterase and the development of antibodies to choline acetyltransferase are two of the major technical advances that have shaped our knowledge of the distribution of central cholinergic neurons. The results, advantages and limitations of both techniques are discussed. A discussion of the phenomenon of coexistence of acetylcholine with neuroactive peptides in central neurons is also included.
Assuntos
Química Encefálica , Neurônios/análise , Sistema Nervoso Parassimpático/análise , Medula Espinal/análise , Acetilcolinesterase/análise , Animais , Córtex Cerebral/análise , Colina O-Acetiltransferase/análise , Corpo Estriado/análise , Hipotálamo/análise , Neurônios Motores/análiseRESUMO
The distribution of immunoreactive (ir)-metorphamide (adrenorphin) in 101 microdissected rat brain and spinal cord regions was determined using a highly specific radioimmunoassay. The highest concentration of metorphamide in brain was found in globus pallidus (280.1 fmol/mg protein). High concentrations of ir-metorphamide (greater than 120 fmol/mg protein) were found in 9 nuclei, including central amygdaloid nucleus, lateral preoptic area, anterior hypothalamic nucleus, hypothalamic paraventricular nucleus, interpeduncular nucleus, periaqueductal grey matter and nucleus of the solitary tract. Moderate concentrations of the peptide (between 60 and 120 fmol/mg protein) were found in 47 brain nuclei such as nucleus accumbens, bed nucleus of stria terminalis, several septal and amygdaloid nuclei, most of the hypothalamic nuclei, ventral tegmental area, red nucleus, raphe nuclei, lateral reticular nucleus, area postrema and others. Low concentrations or ir-metorphamide (less than 60 fmol/mg protein) were measured in 41 nuclei, e.g., cortical structures, hippocampus, caudate nucleus, thalamic nuclei, supraoptic nucleus, substantia nigra, vestibular nuclei, cerebellum (nuclei and cortex). The olfactory bulb has the lowest metorphamide concentration (5.8 fmol/mg protein). Spinal cord segments exhibit very low peptide concentrations.
Assuntos
Química Encefálica , Encefalina Metionina/análogos & derivados , Animais , Cerebelo/análise , Diencéfalo/análise , Encefalina Metionina/análise , Hipotálamo/análise , Masculino , Bulbo/análise , Mesencéfalo/análise , Ponte/análise , Ratos , Ratos Endogâmicos , Medula Espinal/análise , Telencéfalo/análise , Distribuição TecidualRESUMO
The neuroanatomical distribution of bombesin-like immunoreactivity (BLI) in the rat central nervous system was investigated using radioimmunoassay and immunohistochemistry. Whereas cross-reactivity of the bombesin antiserum with substance P was problematic in the immunohistochemical experiments, no significant cross-reactivity with substance P was apparent in the radioimmunoassay. Results from the radioimmunoassay studies reveal particularly high concentrations of BLI in the hypothalamus, thalamus, medulla and spinal cord. Adult rats treated neonatally with capsaicin displayed significant depletions of somatostatin-like and substance P-like immunoreactivity and a small, statistically significant, reduction of BLI in the cervical spinal cord. Capsaicin treatment significantly reduced substance P-like immunoreactivity, but not somatostatin-like immunoreactivity, in the medulla and resulted in a small BLI depletion of borderline statistical significance in this brain region. Neonatally administered capsaicin treatment had no effect on the thalamic concentration of any of these three neuropeptides and neurotensin-like immunoreactivity was unchanged in all brain regions studied. These results suggest that the source of some of the BLI found in the spinal cord may be capsaicin-sensitive dorsal root ganglion cells.