RESUMEN
Traumatic brain injury is a leading cause of hypopituitarism, which compromises patients' recovery, quality of life, and life span. To date, there are no means other than standardized animal studies to provide insights into the mechanisms of posttraumatic hypopituitarism. We have found that GH levels were impaired after inducing a controlled cortical impact (CCI) in mice. Furthermore, GHRH stimulation enhanced GH to lower level in injured than in control or sham mice. Because many characteristics were unchanged in the pituitary glands of CCI mice, we looked for changes at the hypothalamic level. Hypertrophied astrocytes were seen both within the arcuate nucleus and the median eminence, two pivotal structures of the GH axis, spatially remote to the injury site. In the arcuate nucleus, GHRH neurons were unaltered. In the median eminence, injured mice exhibited unexpected alterations. First, the distributions of claudin-1 and zonula occludens-1 between tanycytes were disorganized, suggesting tight junction disruptions. Second, endogenous IgG was increased in the vicinity of the third ventricle, suggesting abnormal barrier properties after CCI. Third, intracerebroventricular injection of a fluorescent-dextran derivative highly stained the hypothalamic parenchyma only after CCI, demonstrating an increased permeability of the third ventricle edges. This alteration of the third ventricle might jeopardize the communication between the hypothalamus and the pituitary gland. In conclusion, the phenotype of CCI mice had similarities to the posttraumatic hypopituitarism seen in humans with intact pituitary gland and pituitary stalk. It is the first report of a pathological status in which tanycyte dysfunctions appear as a major acquired syndrome.
Asunto(s)
Lesiones Encefálicas/fisiopatología , Modelos Animales de Enfermedad , Células Ependimogliales/patología , Hipopituitarismo/etiología , Hipotálamo/patología , Neuronas/patología , Uniones Estrechas/patología , Animales , Núcleo Arqueado del Hipotálamo/inmunología , Núcleo Arqueado del Hipotálamo/metabolismo , Núcleo Arqueado del Hipotálamo/patología , Biomarcadores/metabolismo , Células Ependimogliales/inmunología , Células Ependimogliales/metabolismo , Regulación de la Expresión Génica , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Hormona Liberadora de Hormona del Crecimiento/genética , Hormona Liberadora de Hormona del Crecimiento/metabolismo , Hipopituitarismo/inmunología , Hipopituitarismo/metabolismo , Hipopituitarismo/patología , Hipotálamo/inmunología , Hipotálamo/metabolismo , Inmunoglobulina G/metabolismo , Masculino , Eminencia Media/inmunología , Eminencia Media/metabolismo , Eminencia Media/patología , Ratones , Ratones Transgénicos , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Neuronas/inmunología , Neuronas/metabolismo , Permeabilidad , Proteínas Recombinantes de Fusión/metabolismo , Tercer Ventrículo/inmunología , Tercer Ventrículo/metabolismo , Tercer Ventrículo/patología , Uniones Estrechas/inmunología , Uniones Estrechas/metabolismoRESUMEN
The enzyme type 2 deiodinase (D2) is a major determinant of T3 production in the central nervous system. It is highly expressed in tanycytes, a specialized cell type lining the wall of the third ventricle. During acute inflammation, the expression of D2 in tanycytes is up-regulated by a mechanism that is poorly understood at present, but we hypothesized that cJun N-terminal kinase 1 (JNK1) and v-rel avian reticuloendotheliosis viral oncogene homolog A (RelA) (the 65 kD subunit of NFκB) inflammatory signal transduction pathways are involved. In a mouse model for acute inflammation, we studied the effects of lipopolysaccharide (LPS) on mRNA expression of D2, JNK1, and RelA in the periventricular area (PE) and the arcuate nucleus-median eminence of the hypothalamus. We next investigated LPS-induced D2 expression in primary tanycyte cell cultures. In the PE, the expression of D2 was increased by LPS. In the arcuate nucleus, but not in the PE, we found increased RelA mRNA expression. Likewise, LPS increased D2 and RelA mRNA expression in primary tanycyte cell cultures, whereas JNK1 mRNA expression did not change. Phosphorylation of RelA and JNK1 was increased in tanycyte cell cultures 15-60 minutes after LPS stimulation, confirming activation of these pathways. Finally, inhibition of RelA with the chemical inhibitors sulfasalazine and 4-Methyl-N¹-(3-phenylpropyl)benzene-1,2-diamine (JSH-23) in tanycyte cell cultures prevented the LPS-induced D2 increase. We conclude that NFκB signaling is essential for the up-regulation of D2 in tanycytes during inflammation.
Asunto(s)
Endotoxinas/toxicidad , Inducción Enzimática/efectos de los fármacos , Células Ependimogliales/efectos de los fármacos , Yoduro Peroxidasa/biosíntesis , FN-kappa B/metabolismo , Proteínas del Tejido Nervioso/biosíntesis , Transducción de Señal/efectos de los fármacos , Animales , Núcleo Arqueado del Hipotálamo/citología , Núcleo Arqueado del Hipotálamo/efectos de los fármacos , Núcleo Arqueado del Hipotálamo/inmunología , Núcleo Arqueado del Hipotálamo/metabolismo , Células Cultivadas , Células Ependimogliales/citología , Células Ependimogliales/inmunología , Células Ependimogliales/metabolismo , Femenino , Yoduro Peroxidasa/genética , Yoduro Peroxidasa/metabolismo , Masculino , Eminencia Media/citología , Eminencia Media/efectos de los fármacos , Eminencia Media/inmunología , Eminencia Media/metabolismo , Ratones , Ratones de la Cepa 129 , Núcleos Talámicos de la Línea Media/citología , Núcleos Talámicos de la Línea Media/efectos de los fármacos , Núcleos Talámicos de la Línea Media/inmunología , Núcleos Talámicos de la Línea Media/metabolismo , Proteína Quinasa 8 Activada por Mitógenos/genética , Proteína Quinasa 8 Activada por Mitógenos/metabolismo , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Ratas , Ratas Wistar , Factor de Transcripción ReIA/biosíntesis , Factor de Transcripción ReIA/genética , Factor de Transcripción ReIA/metabolismo , Yodotironina Deyodinasa Tipo IIRESUMEN
Alcoholic beverages are characterized by their fermented versus distilled origin and also by their degree of alcohol. The toxic effects of chronic alcohol consumption have been widely studied. However, there is less evidence about possible beneficial effects of moderate alcohol intake. This work was aimed at evaluating the effects of moderate alcohol consumption (beer or ethanol) on plasma hormone concentrations, blood and thymus lymphocyte phenotypes and brain neurotransmitter levels. For this purpose, 40 adult Wistar male rats were administered ethanol or beer for 4 weeks (experimental groups). Age-matched rats were administered beer without alcohol or water to be used as controls. Rats were killed by decapitation and plasma from the trunk blood was collected to measure plasma prolactin, growth hormone and ACTH concentrations by homologous specific double antibody radioimmunoassays. Thymus and blood lymphocyte subsets were measured by flow cytometry. Neurotransmitter concentrations [dopamine, gamma-aminobutyric acid (GABA) and taurine] were measured by high pressure liquid chromatography in the median eminence and the pituitary. Blood and thymus lymphocyte subsets were not significantly changed by either ethanol or beer consumption, compared to controls. Plasma prolactin levels significantly decreased in ethanol-administered groups (p < 0.05) compared to control animals drinking water, although plasma levels of growth hormone and ACTH were not modified by either alcohol used. Dopamine and GABA concentrations in the median eminence or in the adenohypophysis remained unmodified by moderate beer or ethanol consumption. However, taurine concentration was significantly increased in the pituitary (p < 0.05) in the group drinking ethanol compared to those groups drinking beer with or without alcohol. These data suggest that moderate alcohol consumption may change the regulatory mechanism of prolactin secretion. Whether these modifications have a physiological significance deserves further research.
Asunto(s)
Bebidas Alcohólicas/efectos adversos , Etanol/farmacología , Sistema Hipotálamo-Hipofisario/efectos de los fármacos , Sistema Inmunológico/efectos de los fármacos , Neuroinmunomodulación/efectos de los fármacos , Prolactina/efectos de los fármacos , Animales , Depresores del Sistema Nervioso Central/farmacología , Dopamina/metabolismo , Sistema Hipotálamo-Hipofisario/inmunología , Sistema Hipotálamo-Hipofisario/metabolismo , Sistema Inmunológico/inmunología , Linfocitos/efectos de los fármacos , Linfocitos/inmunología , Masculino , Eminencia Media/efectos de los fármacos , Eminencia Media/inmunología , Eminencia Media/metabolismo , Neuroinmunomodulación/inmunología , Adenohipófisis/efectos de los fármacos , Adenohipófisis/inmunología , Adenohipófisis/metabolismo , Prolactina/sangre , Prolactina/metabolismo , Ratas , Ratas Wistar , Taurina/metabolismo , Timo/citología , Timo/efectos de los fármacos , Timo/inmunología , Ácido gamma-Aminobutírico/metabolismoRESUMEN
CONTEXT: Autoantibodies to adenohypophyseal endocrine cells or to vasopressin neurohypophyseal neurons have long been known. Conversely, autoimmune targeting of further hypothalamic-hypophyseal structures, such as the blood-brain barrier-deprived median eminence, has been little studied. OBJECTIVE AND METHODS: We studied a case of autoimmune polyendocrine syndrome type I with GH secretory deficiency, a distinctly rare event in autoimmune polyendocrine syndrome type I. We used rat and bovine tissue substrates to study autoantibodies against hypothalamic-hypophyseal nerve structures and endocrine cells. RESULTS: In the study case, circulating autoantibodies selectively decorated median eminence dopaminergic nerve terminals, as well as pituitary gonadotropes, but not GHRH nerve terminals or pituitary somatotropes. Such autoantibodies appeared de novo in parallel with the onset of GH secretory deficiency, whereas no median eminence labeling was found in patients suffering of idiopathic GH deficiency (n = 7) or in healthy controls (n = 23). CONCLUSIONS: The pathophysiological significance of our patient's autoantibodies remains to be confirmed. Nonetheless, the heterogeneous neuroendocrine structures of the median eminence are pointed out as potential immune targets, relevant to autoimmune polyendocrinopathy, as well as to a wide range of other conditions.
Asunto(s)
Eminencia Media/inmunología , Poliendocrinopatías Autoinmunes/inmunología , Adolescente , Animales , Descarboxilasas de Aminoácido-L-Aromático/inmunología , Autoanticuerpos/sangre , Bovinos , Niño , Preescolar , Femenino , Hormona Liberadora de Gonadotropina/inmunología , Hormona de Crecimiento Humana/deficiencia , Hormona de Crecimiento Humana/metabolismo , Humanos , Masculino , Ratas , Ratas Sprague-Dawley , Tirosina 3-Monooxigenasa/inmunologíaRESUMEN
We analyzed expression of osteopontin (OPN), a cytokine regulating tissue repair and inflammation, in astrocytes and microglia in response to systemic lipopolysaccharide (LPS) administration (250 microg/100 g). OPN mRNA expression appeared in subpial astrocytes as early as 6 h, and then spread over the brain parenchyma. The signal for OPN mRNA reached a peak at 24 h post-injection, and returned to basal levels after 48 h. Changes in OPN immunoreactivity in the LPS-injected rat mirrored OPN mRNA induction patterns. These results provide the first evidence of OPN induction in astrocytes and microglia following peripheral immune challenge, and suggest that OPN may play a key role in the pathogenesis of neuroinflammation.
Asunto(s)
Encéfalo/inmunología , Encéfalo/metabolismo , Regulación de la Expresión Génica/inmunología , Lipopolisacáridos/administración & dosificación , ARN Mensajero/biosíntesis , Sialoglicoproteínas/biosíntesis , Sialoglicoproteínas/genética , Animales , Núcleo Arqueado del Hipotálamo/inmunología , Núcleo Arqueado del Hipotálamo/metabolismo , Núcleo Arqueado del Hipotálamo/ultraestructura , Encéfalo/ultraestructura , Hipotálamo/inmunología , Hipotálamo/metabolismo , Hipotálamo/ultraestructura , Inmunohistoquímica , Hibridación in Situ , Inyecciones Intraperitoneales , Masculino , Eminencia Media/inmunología , Eminencia Media/metabolismo , Eminencia Media/ultraestructura , Microscopía Inmunoelectrónica , Osteopontina , Ratas , Ratas Sprague-Dawley , Sialoglicoproteínas/ultraestructura , Núcleo Supraóptico/inmunología , Núcleo Supraóptico/metabolismo , Núcleo Supraóptico/ultraestructura , Factores de TiempoRESUMEN
Tanycytes are specialized ependymal cells lining the infundibular recess of the third ventricle of the cerebrum. Early and recent investigations involve tanycytes in the mechanism of gonadotropin-releasing hormone (GnRH) release to the portal blood. The present investigation was performed to obtain a specific immunological marker of tanycytes and to identify the compound(s) responsible for this labeling. After 30 days of organ culture, explants of bovine median eminence formed spherical structures mostly constituted by tanycytes. These tanycyte spheres were xenotransplanted to rats, and the antibodies raised by the host animals against the transplanted living tanycytes were used for immunochemical studies of the bovine and rat median eminence. This antiserum immunoreacted with two compounds of 60 kDa and 85 kDa present in extracts of bovine and rat median eminence. The individual immunoblotting analysis of rat medial basal hypothalami showed a decrease in the amount of the 85-kDa compound in castrated rats as compared to control rats processed at oestrus and dioestrus. The antiserum, labeled as anti-P85, when used for immunostaining of sections throughout the rat central nervous system, immunoreacted specifically with the hypothalamic tanycytes. Within tanycytes, P-85 immunoreactivity was exclusively present in the basal processes. It is suggested that the 85-kDa and 60-kDa compounds correspond to two novel proteins selectively expressed by tanycytes. The possibility that they are secretory proteins involved in GnRH release is discussed. Anti-P85 appears to be the first specific marker of hypothalamic tanycytes.
Asunto(s)
Hipotálamo/citología , Hipotálamo/inmunología , Eminencia Media/inmunología , Eminencia Media/trasplante , Trasplante Heterólogo/inmunología , Animales , Bovinos , Femenino , Hipotálamo/ultraestructura , Immunoblotting , Inmunohistoquímica , Masculino , Microscopía Electrónica , Proteínas del Tejido Nervioso/metabolismo , Técnicas de Cultivo de Órganos , Ratas , Ratas Sprague-Dawley , Extractos de Tejidos/inmunología , Vacuolas/ultraestructuraRESUMEN
Lewis rats exhibit multiple defects in their hypothalamus-pituitary-adrenal (HPA) system that are considered to play a causal role in the susceptibility of this strain to autoimmune diseases, i.e. experimental allergic encephalomyelitis (EAE). In the present study, we aimed to modulate the HPA response of the Lewis rat and establish its consequences for the susceptibility to EAE. Because in Wistar rats, single administration of interleukin (IL)-beta (priming) is known to induce long-lasting (weeks) sensitization of HPA responses to stressors and immune stimuli, Lewis rats were given a single dose of hIL-1beta or vehicle 1 week prior to induction of EAE by immunization with myelin basic protein (MBP). Subsequently, neurological deficits were monitored once daily. The results show that IL-1 priming markedly suppresses the neurological symptoms of EAE, without affecting the onset or duration of the disease. Measurement of vasopressin and corticotropin releasing hormone (CRH) in the external zone of the median eminence revealed that, as compared to Wistar rats, Lewis rats exhibit low vasopressin but identical CRH, and that IL-1 priming increases (0.001) vasopressin without affecting CRH stores, which is consistent with a shift to vasopressin-dominated control of adrenocorticotropic hormone (ACTH) secretion as described in Wistar rats under conditions of HPA hyper(re)activity. However, IL-1 priming did not affect a.m. corticosterone levels following immunization with MBP or during the clinical phase of EAE. IL-1 priming of Lewis rats attenuated the ACTH responses to an IL-1 challenge 11 days later, which may relate to an increase in resting corticosterone levels. Thus, the mechanisms underlying IL-1 induced suppression of EAE are not related to enhanced HPA responses. In addition, we did not find IL-1 priming-induced alterations in MBP-specific immunoglobulin (Ig)M, IgG1, IgGa and IgGb plasma titres, or gross alterations in T cell activation as reflected in spontaneous or concanavalin-induced T cell proliferation. We therefore speculate that IL-1-induced elevation of resting corticosterone levels may influence the development of EAE.
Asunto(s)
Encefalomielitis Autoinmune Experimental/tratamiento farmacológico , Encefalomielitis Autoinmune Experimental/inmunología , Interleucina-1/farmacología , Hormona Adrenocorticotrópica/sangre , Animales , División Celular/efectos de los fármacos , División Celular/inmunología , Concanavalina A/farmacología , Corticosterona/sangre , Hormona Liberadora de Corticotropina/análisis , Hormona Liberadora de Corticotropina/inmunología , Encefalomielitis Autoinmune Experimental/inducido químicamente , Inmunización , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Masculino , Eminencia Media/química , Eminencia Media/efectos de los fármacos , Eminencia Media/inmunología , Proteína Básica de Mielina/inmunología , Proteína Básica de Mielina/farmacología , Examen Neurológico , Ratas , Ratas Endogámicas Lew , Ratas Wistar , Linfocitos T/citología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Vasopresinas/análisis , Vasopresinas/inmunologíaRESUMEN
The effect of Freund's adjuvant injection on 24-h variation of circulating ACTH, prolactin, growth hormone (GH), and thyroid-stimulating hormone (TSH) levels, and of norepinephrine (NE) content, and dopamine (DA) and serotonin (5HT) turnover in median eminence, was examined in adult rats kept under light between 0800 and 2000 h daily. Groups of 6-10 animals received Freund's complete adjuvant or its vehicle at 1100 h 3 days before sacrifice and were killed by decapitation at six different time intervals throughout a 24-h cycle. In rats injected with adjuvant's vehicle, serum ACTH and prolactin exhibited peak values around the light-dark transition (p < 0.0001 and < 0.04, respectively), while the maximum in TSH was found in the late afternoon (p < 0.0001, one-way ANOVA). GH levels did not vary on a 24-h basis. In Freund's adjuvant-injected rats, 24-h variations of TSH levels became blunted, while 24-h variations of prolactin and ACTH persisted. Freund's adjuvant augmented serum ACTH and prolactin levels, and decreased GH and TSH levels (p < 0.0007, factorial ANOVA). Median-eminence NE content, and turnover of DA, assessed by measuring dihydroxyphenylacetic acid, DOPAC/DA ratio, and of 5HT, assessed by measuring 5-hydroxyindoleacetic acid, HIAA/5HT ratio, varied on a 24-h basis in rats receiving adjuvant's vehicle (p < 0.02). Median-eminence NE content attained its maximum at 1600-2000 h, while maxima in DOPA/DA and HIAA/5HT ratios occurred at 0400 h. Injection with Freund's adjuvant reduced the amplitude of the daily variation of NE content, shifted the maximum of DOPAC/DA ratio toward the light-dark transition, and blunted the daily variation in HIAA/5HT ratio in median eminence. The administration at 1200 of the immunosuppressant drug cyclosporine (5 mg/kg, 5 days) restored the augmented ACTH and prolactin levels (p < 0.0001, factorial ANOVA) and depressed GH and TSH levels (p < 0.02) found in Freund's adjuvant-injected rats. Cyclosporine was also effective in restoring 24-h rhythmicity of serum ACTH and TSH, but not of prolactin, levels. Cyclosporine did not modify the effect of Freund's adjuvant on time-of-day changes of median-eminence NE content, but it was effective in counteracting the changes of DA and 5HT turnover found after immunization. The results are compatible with a significant effect of immune-mediated inflammatory response at an early phase after Freund's adjuvant injection on ACTH, GH, prolactin, and TSH release, which is partially sensitive to immunosuppression by cyclosporine.
Asunto(s)
Artritis Experimental/sangre , Artritis Experimental/metabolismo , Monoaminas Biogénicas/metabolismo , Ritmo Circadiano/fisiología , Hormonas/sangre , Eminencia Media/metabolismo , Ácido 3,4-Dihidroxifenilacético/metabolismo , Hormona Adrenocorticotrópica/sangre , Animales , Artritis Experimental/etiología , Ritmo Circadiano/inmunología , Dopamina/metabolismo , Adyuvante de Freund/administración & dosificación , Hormona del Crecimiento/sangre , Ácido Hidroxiindolacético/metabolismo , Masculino , Eminencia Media/inmunología , Norepinefrina/metabolismo , Prolactina/sangre , Ratas , Ratas Wistar , Serotonina/metabolismo , Tirotropina/sangreRESUMEN
A system of intraventricular neuronal perikarya and processes known as the supraependymal neuronal complex (SENC) is located on the floor of the third ventricle and innervates the neurohypophysis of the golden hamster (Mesocricetus auratus). Immunocytochemical techniques were used to determine if oxytocin and/or vasopressin are present in the neuronal elements of the SENC. Oxytocinergic fibers were observed to breach the ependyma of the median eminence and enter the neuropil of the SENC. Some of these fibers traverse the SENC to reach the adjacent ependymal surface and terminate on the floor of the third ventricle while others terminate within the neuropil of the SENC. These oxytocinergic fibers may be involved in the secretion of oxytocin into the cerebrospinal fluid (CSF). Vasopressinergic fibers were detected in the neuropil of the SENC in only one of four specimens examined and are assumed to be aberrant processes from the hypothalamo-neurohypophysial tract. Neither oxytocin nor vasopressin were detected in the neurons intrinsic to the SENC. The function of the SENC is unknown, but it may be involved in regulatory processes in which CSF oxytocin has been implicated, such as osmotic homeostasis and/or cardiovascular reflexes.
Asunto(s)
Ventrículos Cerebrales/metabolismo , Neuronas/metabolismo , Oxitocina/metabolismo , Animales , Ventrículos Cerebrales/citología , Cricetinae , Femenino , Inmunohistoquímica , Masculino , Eminencia Media/citología , Eminencia Media/inmunología , Eminencia Media/metabolismo , Mesocricetus , Neuronas/inmunología , Oxitocina/inmunología , Vasopresinas/inmunología , Vasopresinas/metabolismoRESUMEN
The terminals of the hypothalamic gonadotrophin hormone-releasing hormone (GnRH) neurons are located within the median eminence and thereby extend beyond the protection of the blood-brain barrier. Thus, these terminals may be subjected to direct autoimmune action in animals that are actively immunised against GnRH. Boars (male pigs) (n = 108) were actively immunised against GnRH by two successive injections with synthetic GnRH, covalently coupled to KLH and dissolved in CFA or IFA. They were killed at 26 weeks of age. Immunised boars were selected on the basis of the resultant testes size, which indicates the effectiveness of the immunisation. The hypothalami of 25 selected animals were studied by histological and immunocytochemical techniques and compared with the hypothalami of three sham- and nine control animals. In the immunised animals, changes in the GnRH system had taken place. These comprised dystrophy of the perikarya and a sharp decrease of the GnRH immunocytochemical reactivity in the terminals within the median eminence. In addition, various degrees of inflammatory reactions were present, particularly within the median eminence. These consisted of tissue disruption by edema, collapse of the capillaries, fibrosis and infiltration with fibroblasts. In addition, accumulations of neurosecretum within the median eminence in combination with hypertrophy of magnocellular neurons within the hypothalamus were present. The reactions were restricted to the median eminence and did not involve other neurohemal organs or other parts of the hypothalamus. A correlation could be established between the incidence of the lesions and the effectiveness of the GnRH autoimmunity (as indicated by the size and endocrine function of the gonads and the anti-GnRH titres). Changes in extra- and intracellular IgG immunocytochemical reactivity within the median eminence indicated the involvement of IgG. The effects were absent from control and sham vaccinated animals and after vaccinations with other compositions of the vaccine. Thus, hypothalamic lesions have been observed in this selected group of animals, vaccinated against GnRH with this particular vaccine.
Asunto(s)
Autoanticuerpos/inmunología , Hormona Liberadora de Gonadotropina/inmunología , Eminencia Media/patología , Secuencia de Aminoácidos , Animales , Proteína Ácida Fibrilar de la Glía/análisis , Proteína Ácida Fibrilar de la Glía/inmunología , Hormona Liberadora de Gonadotropina/análisis , Inmunoglobulina G/inmunología , Masculino , Eminencia Media/inmunología , Datos de Secuencia Molecular , Porcinos , VacunaciónRESUMEN
The enkephalin-immunoreactive neurons that project to the external zone of the median eminence were identified on thin paraffin and thick vibratome sections using a combination of retrograde labeling with peripherally administered Fluoro-Gold and immunocytochemistry. The vast majority of the enkephalin-immunoreactive neurons that project to the external zone of the median eminence (ME) reside in the paraventricular nucleus (PVN) of the hypothalamus. Within the PVN, the majority of these hypophysiotropic neurons are located in the medial parvicellular subdivision, while a smaller number can be detected in the anterior and the periventricular subdivisions. Although many enkephalin-immunoreactive neurons are present in other hypophysiotropic areas of the hypothalamus, such as the medial preoptic area, the anterior periventricular area, and the arcuate nucleus, only a few of these can be retrogradely labeled from the ME. These results provide morphological evidence for the key role of paraventricular enkephalin-immunoreactive neurons in the regulation of neuroendocrine functions.
Asunto(s)
Encefalinas/metabolismo , Eminencia Media/citología , Neuronas/metabolismo , Núcleo Hipotalámico Paraventricular/citología , Estilbamidinas , Animales , Encefalinas/inmunología , Colorantes Fluorescentes , Inmunohistoquímica , Masculino , Eminencia Media/inmunología , Vías Nerviosas/citología , Neuronas/inmunología , Núcleo Hipotalámico Paraventricular/inmunología , Ratas , Ratas Sprague-DawleyRESUMEN
Immunoreactive galanin-like material was recently shown to co-exist with vasopressin in parvocellular and magnocellular perikarya of the paraventricular nucleus in the anterior hypothalamus of the rat (Melander et al. 1986). Since this distribution pattern differed from our observation of oxytocin-associated galanin-like immunoreactivity (LI) in the neurohypophysis, we compared in series of 0.5-microns thick sections the localisation of galanin-LI with the localisation of oxytocin and vasopressin/dynorphin in the hypothalamus, the median eminence and the neurohypophysis. In the oxytocin system, galanin-LI was intense in oxytocin varicosities of the neurohypophysis. Oxytocin perikarya of the hypothalamic supraoptic and paraventricular nuclei exhibited galanin-LI only after intraventricular injection of colchicine and when sections were treated with trypsin prior to application of the antibody. In the vasopressin/dynorphin system galanin-LI was intense in hypothalamic perikarya after colchicine injection and in neurohypophysial varicosities after treatment of the sections with trypsin. In these neurones, galanin-LI was absent or weak in all elements when treatments with colchicine or trypsin were omitted. Galanin-LI in the neurohypophysis was not co-localised with the numerous fine endings showing GABA-LI. These observations indicate that galanin-like material coexists with vasopressin and oxytocin in the respective magnocellular neurones, although not always in an immunoreactive form.
Asunto(s)
Hipotálamo/inmunología , Neuronas/inmunología , Péptidos/inmunología , Neurohipófisis/inmunología , Animales , Catecolaminas/metabolismo , Femenino , Galanina , Hipotálamo/citología , Hipotálamo/metabolismo , Inmunohistoquímica , Masculino , Eminencia Media/citología , Eminencia Media/inmunología , Eminencia Media/metabolismo , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Péptidos/metabolismo , Neurohipófisis/citología , Neurohipófisis/metabolismo , Ratas , Ratas Endogámicas , Tripsina/farmacologíaRESUMEN
The distribution and molecular forms of bombesin-like immunoreactivity (BLI) were determined in the ovine median eminence using a new, C-terminally directed antiserum. BLI was confined to neurons of the external zone of the median eminence, near small blood vessels, many of which also were immunoreactive for CRF. Each median eminence contained about 10 pmol of BLI (533 pmol/g tissue). Gel filtration and reverse-phase high pressure liquid chromatography (HPLC) demonstrated the existence of two molecular forms of BLI, which co-eluted with porcine gastrin releasing peptide (GRP)1-27 and GRP18-27 in a molar ratio of 1:2. The presence of BLI in the ovine median eminence and its co-localization in some neurons with CRF, suggest a possible role for peptides of the bombesin family in the regulation of pituitary function.
Asunto(s)
Bombesina/análisis , Eminencia Media/inmunología , Animales , Cromatografía Líquida de Alta Presión , Sueros Inmunes , Inmunohistoquímica , Conformación Molecular , Fragmentos de Péptidos/análisis , Radioinmunoensayo , OvinosRESUMEN
An antiserum was raised to adipokinetic hormone (AKH), a 10-amino-acid-residue peptide found in the arthropod Locusta migratoria. The antiserum demonstrated not only immunocytochemical reaction with some other arthropod species, but also stained many areas of the rat CNS, certain islet cells of the pancreas, and some anterior pituitary cells. The pattern of staining was unlike that for any known rat neuropeptide or hormone. With the antiserum used as the detection system, HPLC and high-voltage electrophoresis yielded two peptides that were purified to homogeneity from rat hypothalamic median eminence. These peptides have unique amino acid compositions, indicating they may be heretofore unknown rat neuropeptides.
Asunto(s)
Hormonas de Insectos/inmunología , Péptidos/inmunología , Ratas/inmunología , Animales , Carcinoma/inmunología , Carcinoma/patología , Línea Celular , Cromatografía Líquida de Alta Presión , Histocitoquímica , Sueros Inmunes/inmunología , Inmunoquímica , Eminencia Media/inmunología , Páncreas/inmunología , Péptidos/aislamiento & purificación , Neoplasias de la Tiroides/inmunología , Neoplasias de la Tiroides/patologíaRESUMEN
Confrontations of immuno-stainings as well as sequential stainings with corticoliberin and vasopressin antisera were performed on contiguous sections of human median eminence (ME) or hypothalamic paraventricular nucleus (PVN). Results show that a double immunoreactivity to both IS is expressed in some parvicellular PVN perikarya, and suggest that such a double immunoreactivity can be possible in some ME fibers. Similar results had been previously obtained in animals only under experimental conditions.
Asunto(s)
Hormona Liberadora de Corticotropina/inmunología , Hipotálamo/citología , Neuronas/inmunología , Vasopresinas/inmunología , Técnica del Anticuerpo Fluorescente , Humanos , Sueros Inmunes/inmunología , Eminencia Media/inmunología , Núcleo Hipotalámico Paraventricular/inmunologíaRESUMEN
Concentrations of dynorphin and alpha-neo-endorphin were measured in rat supraoptic and paraventricular nuclei, median eminence and posterior pituitary. Paraventricular lesions did not alter the level of either peptide in the median eminence or posterior pituitary. Knife cuts in the lateral retrochiasmatic area that severed the supraoptico-hypophyseal tract reduced dynorphin and alpha-neo-endorphin levels by 64-80% in the posterior pituitary and by somewhat less in the median eminence. Knife cuts in the vicinity of the supraoptic nucleus that did not interrupt the supraoptico-hypophyseal tract did not affect peptide concentrations in the posterior pituitary. Our data suggest that dynorphin and alpha-neo-endorphin in the posterior pituitary are in processes of supraoptic nucleus neurons. These neurons also project to the median eminence which may receive fibers from other dynorphin and alpha-neo-endorphin containing cells too.
Asunto(s)
Endorfinas/inmunología , Sistema Hipotálamo-Hipofisario/inmunología , Precursores de Proteínas/inmunología , Ratas/fisiología , Animales , Dinorfinas , Masculino , Eminencia Media/inmunología , Neurohipófisis/inmunología , Ratas Endogámicas , Distribución TisularRESUMEN
The zona externa of the median eminence was found completely depleted of corticoliberin-like immunoreactivity (CLI) and of vasopressin-like immunoreactivity (VLI) 12-14 h and 22-24 h after bilateral adrenalectomy. Amount of CLI increased to the level of untreated rats 5 days after this operation while amount of VLI rose higher than in control animals. The same was found 20 days after surgery. These results emphasize the involvement of two infundibular neurosecretory systems--CLI and VLI--in the regulation of the pituitary--adrenal axis.
Asunto(s)
Adrenalectomía , Hormona Liberadora de Corticotropina/inmunología , Eminencia Media/inmunología , Ratas/inmunología , Vasopresinas/inmunología , Animales , Histocitoquímica , Técnicas Inmunológicas , Masculino , Ratas Endogámicas , Factores de TiempoRESUMEN
A new immunohistological method was elaborated for the light-microscopic detection of two tissue antigens in the same section. One of the antigens was visualized by the silver-gold intensified peroxidase-antiperoxidase complex (PAP)-diaminobenzidine (DAB) method resulting in a black coloured end-product. The other antigen was labelled by the brown reaction-product of the traditional PAP-DAB method. Both reaction-products are stable, electron-dense and they can clearly be distinguished from each other.
Asunto(s)
Antígenos/análisis , Técnicas para Inmunoenzimas , Eminencia Media/inmunología , Adenohipófisis/inmunología , Animales , Femenino , Hormona Folículo Estimulante/análisis , Hormona Liberadora de Gonadotropina/análisis , Hormona del Crecimiento/análisis , Eminencia Media/análisis , Adenohipófisis/análisis , Ratas , Plata , Somatostatina/análisis , Coloración y EtiquetadoRESUMEN
With the use of rabbit anti-LH-RH serum, a system of delicate nerve fibers and terminals which contain immunoreactive LH-RH can be demonstrated in the median eminence of the rat, corresponding exactly to the tubero-infundibular tract as described earlier with the aid of classical neurophistological methods. The delicate fiber system can be traced to the region of the rectrochiasmatic area and especially to that of the arcuate nucleus but not farther. Nerve cells do not show any immunopositive reaction for LH-RH in the experimental conditions studied. LH-RH positive nerve fibers terminate on or in the immediate vicinity of the capillary loops penetrating the median eminence.