RESUMEN
BACKGROUND: sensitivity to food antigens has been postulated as a contributing factor to the pathogenesis of IgA nephropathy (IgAN). METHODS: in this study we used a recently developed mucosal patch technique to evaluate rectal mucosal sensitivity to soy and cow's milk (CM) proteins in IgAN patients (n = 28) compared to healthy subjects (n = 18). The rectal mucosal production of nitric oxide (NO) and release of myeloperoxidase (MPO) and eosinophil cationic protein (ECP) were measured. Serum samples were analyzed for IgA and IgG antibodies to alpha-lactalbumin, beta-lactoglobulin, casein and soy. RESULTS: 14 of 28 (14/28) patients experienced a rectal mucosal reaction, measured by increased NO and/or MPO levels, upon rectal challenge with soy and/or cow's milk proteins. The levels of IgG antibodies to alpha-lactalbumin, beta-lactoglobulin and casein were significantly higher in CM sensitive as compared with non-sensitive IgAN patients, whereas the mean serum levels of IgA antibodies were similar. No differences were seen in serum levels of IgA or IgG antibodies to soy. CONCLUSION: it is concluded that approximately half of our IgAN patients have a rectal mucosal sensitivity to soy or CM, and that an immune reactivity against antigens may be involved in the pathogenesis of IgAN in this subgroup of patients.
Asunto(s)
Hipersensibilidad a los Alimentos/epidemiología , Glomerulonefritis por IGA/epidemiología , Mucosa Intestinal/inmunología , Hipersensibilidad a la Leche/epidemiología , Proteínas de la Leche/efectos adversos , Proteínas de Soja/efectos adversos , Inmunidad Adaptativa , Adulto , Anciano , Estudios de Casos y Controles , Caseínas/efectos adversos , Proteína Catiónica del Eosinófilo/metabolismo , Femenino , Hipersensibilidad a los Alimentos/inmunología , Glomerulonefritis por IGA/inmunología , Glomerulonefritis por IGA/fisiopatología , Humanos , Inmunidad Innata , Inmunidad Mucosa , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Pruebas Inmunológicas , Riñón/fisiopatología , Lactalbúmina/efectos adversos , Lactoglobulinas/efectos adversos , Masculino , Persona de Mediana Edad , Hipersensibilidad a la Leche/inmunología , Proteínas de la Leche/inmunología , Óxido Nítrico/metabolismo , Peroxidasa/metabolismo , Proteinuria/epidemiología , Recto , Proteínas de Soja/inmunología , Suecia/epidemiología , Factores de Tiempo , Adulto JovenRESUMEN
OBJECTIVES: A dietary link to rheumatoid arthritis (RA) has been suspected and an influence on arthritic symptoms by different diets has been reported. Our primary aim was to record the self-experienced adverse food reactions in patients with RA. A secondary aim was to relate self-experienced adverse reactions to dairy produce and wheat to the local mucosal reactivity observed after rectal challenge with cow's milk protein (CM) and wheat gluten. METHODS: A questionnaire about self-experienced adverse reaction to food was sent to 347 RA patients. Rectal challenge with CM and gluten was performed in 27 of these patients and in healthy controls (n = 18). After a 15-h challenge the mucosal production of nitric oxide (NO) and the mucosal release of myeloperoxidase (MPO) and eosinophil cationic protein (ECP) were measured by using the mucosal patch technique. RESULTS: Twenty-seven per cent of the RA patients reported food intolerance (FI) to various foods, and in particular to CM, meat, and wheat gluten. Strong mucosal reactivity to CM was observed in 11% of the patients. Moderately increased mucosal reactivity to CM and gluten was found in 22% and 33%, respectively, of the patients. No relationship was found between self-experienced adverse reactions to CM or gluten and mucosal reactivity to these proteins. CONCLUSIONS: Perceived FI is reported frequently by RA patients, with a prevalence similar to that reported previously in the general population. Mucosal reactivity to CM and gluten is seen in a minor fraction of RA patients and is not related to the frequently perceived intolerance to these proteins.
Asunto(s)
Artritis Reumatoide/inmunología , Glútenes/efectos adversos , Hipersensibilidad a la Leche/inmunología , Proteínas de la Leche/efectos adversos , Recto/inmunología , Hipersensibilidad al Trigo/inmunología , Administración Rectal , Adulto , Artritis Reumatoide/complicaciones , Artritis Reumatoide/metabolismo , Femenino , Glútenes/administración & dosificación , Glútenes/inmunología , Glútenes/metabolismo , Humanos , Inmunoglobulina A/inmunología , Inmunoglobulina A/metabolismo , Inmunoglobulina G/inmunología , Inmunoglobulina G/metabolismo , Masculino , Persona de Mediana Edad , Hipersensibilidad a la Leche/complicaciones , Hipersensibilidad a la Leche/metabolismo , Proteínas de la Leche/administración & dosificación , Proteínas de la Leche/inmunología , Proteínas de la Leche/metabolismo , Membrana Mucosa/inmunología , Membrana Mucosa/metabolismo , Óxido Nítrico/metabolismo , Pruebas del Parche , Peroxidasa/metabolismo , Recto/metabolismo , Índice de Severidad de la Enfermedad , Estadísticas no Paramétricas , Encuestas y Cuestionarios , Hipersensibilidad al Trigo/complicaciones , Hipersensibilidad al Trigo/metabolismoRESUMEN
INTRODUCTION: Patients with primary Sjögren's syndrome (pSS) are reported to have a variety of gastrointestinal symptoms partly attributed to an overrepresentation of celiac disease. We have observed that irritable bowel syndrome (IBS)-like symptoms are frequent complaints in this patient group. Allergic manifestations to various drugs are also common in pSS. A role of food allergy in IBS has been proposed. OBJECTIVE: This study is aimed at evaluating the mucosal response to rectal challenge with cow's milk protein (CM) in patients with pSS and relates possible CM reactivity to their intestinal symptoms. METHODS: A rectal challenge with CM was performed in 21 patients with pSS and 18 healthy controls. Fifteen hours after challenge the mucosal production of nitric oxide (NO) and the release of myeloperoxidase (MPO) as signs of mucosal inflammatory reaction were measured using the mucosal patch technique. RESULTS: Eight out of 21 patients with pSS had a definite increase of mucosal NO synthesis and the luminal release of MPO after rectal CM challenge. This sign of milk sensitivity was not linked to IgG/IgA antibodies to milk proteins. The symptoms for IBS according to Rome III criteria were fulfilled in 13 patients. All patients who were CM sensitive suffered from IBS. In a small open study, patients reactive to CM reported an improvement of intestinal symptoms on a CM-free diet. CONCLUSION: A rectal mucosal inflammatory response after CM challenge is seen in 38% of patients with pSS as a sign of CM sensitivity. IBS-like symptoms were common in pSS, linked to CM sensitivity.
Asunto(s)
Síndrome del Colon Irritable/complicaciones , Hipersensibilidad a la Leche/complicaciones , Proteínas de la Leche/efectos adversos , Síndrome de Sjögren/complicaciones , Adulto , Anciano , Animales , Bovinos , Femenino , Antígenos HLA-DQ/análisis , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Síndrome del Colon Irritable/inmunología , Masculino , Persona de Mediana Edad , Hipersensibilidad a la Leche/diagnóstico , Hipersensibilidad a la Leche/inmunología , Membrana Mucosa/inmunología , Membrana Mucosa/metabolismo , Óxido Nítrico/metabolismo , Pruebas del Parche/métodos , Peroxidasa/metabolismo , Recto/inmunología , Recto/metabolismo , Síndrome de Sjögren/inmunología , Estadísticas no ParamétricasRESUMEN
BACKGROUND: Palmoplantar pustulosis (PPP) is a chronic inflammatory disease affecting mainly smoking women. Some patients also have psoriasis. A subgroup of patients with psoriasis has been shown to have silent gluten sensitivity with relevance for their psoriasis. Nothing is known about gluten sensitivity in PPP. OBJECTIVES: To find out whether any patients with PPP are gluten-sensitive and whether this might be relevant for the PPP activity. PATIENTS AND METHODS: One hundred and twenty-three patients (113 women) with PPP participated. Screening for IgA antibodies against gliadin and tissue transglutaminase (tTG) was performed, the duodenal mucosa in patients with and without these antibodies was studied and the effect of a gluten-free diet (GFD) was followed up. RESULTS: Twenty-two patients (18%) had IgA antibodies against gliadin and nine of 94 (10%) against tTG. Twelve patients with antibodies and 11 without underwent gastro-duodenoscopy. Four displayed villous atrophy, whereas all other specimens were judged as essentially normal at routine staining. However, with immunohistochemistry, the numbers of CD3+ and CD8+ lymphocytes in the epithelium were found to be increased in patients with any type of antibody, although they were most numerous in those with both types of antibodies. Seven of 123 patients (6%) had coeliac disease (three previously diagnosed). Patients with antibodies who adhered to the GFD displayed total or nearly total clearance of the skin lesions and normalization of the antibody levels. CONCLUSIONS: Patients with PPP should be screened for antibodies against gliadin and tTG. Those with antibodies can be much improved on a GFD regardless of the degree of mucosal abnormalities.
Asunto(s)
Gliadina/inmunología , Inmunoglobulina G/sangre , Psoriasis/enzimología , Psoriasis/inmunología , Transglutaminasas/inmunología , Hipersensibilidad al Trigo/prevención & control , Adulto , Dieta con Restricción de Proteínas , Femenino , Humanos , Mucosa Intestinal/inmunología , Masculino , Persona de Mediana Edad , Psoriasis/sangre , Psoriasis/diagnóstico , Resultado del TratamientoRESUMEN
Patients with coeliac disease (CD) on a gluten-free diet may still have gastrointestinal symptoms. On clinical grounds cow's milk (CM) protein sensitivity may be suspected. Here, using rectal protein challenge, we investigated the local inflammatory reaction to gluten and CM protein in adult patients with CD in remission. Rectal challenges with wheat gluten and dried CM powder were performed in 20 patients with CD and 15 healthy controls. Fifteen hours after challenge the mucosal reaction was recorded by the mucosal patch technique with measurements of local release of neutrophil and eosinophil granule constituents; myeloperoxidase (MPO) and eosinophil cationic protein (ECP). We measured the mucosal production of nitric oxide (NO) simultaneously. Six of the patients who reacted to CM were also challenged with alpha-lactalbumin and casein. In 18 of 20 patients gluten challenge induced neutrophil activation defined as increased MPO release and increased NO synthesis. Ten of these 20 patients showed a similarly strong inflammatory reaction to CM challenge. Six of the CM sensitive patients were challenged with specific CM proteins: casein and alpha-lactalbumin. Casein, in contrast to alpha-lactalbumin, induced an inflammatory response similar to that produced by CM. A mucosal inflammatory response similar to that elicited by gluten was produced by CM protein in about 50% of the patients with coeliac disease. Casein, in particular, seems to be involved in this reaction.
Asunto(s)
Enfermedad Celíaca/etiología , Hipersensibilidad a la Leche/complicaciones , Proteínas de la Leche/inmunología , Administración Rectal , Adulto , Anciano , Animales , Caseínas/inmunología , Bovinos , Enfermedad Celíaca/inmunología , Femenino , Glútenes/administración & dosificación , Glútenes/inmunología , Humanos , Inmunidad Mucosa , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Mucosa Intestinal/inmunología , Lactalbúmina/inmunología , Masculino , Persona de Mediana Edad , Hipersensibilidad a la Leche/diagnóstico , Hipersensibilidad a la Leche/inmunología , Proteínas de la Leche/administración & dosificación , Óxido Nítrico/metabolismo , Peroxidasa/metabolismoRESUMEN
BACKGROUND AND AIMS: To elucidate the dynamics of nitric oxide (NO) production induced by rectal gluten challenge and the relation between NO production and mucosal granulocyte activation. SUBJECTS AND METHODS: Release of rectal NO was measured in 13 patients with coeliac disease and in 18 controls before and after rectal wheat gluten challenge. Rectal gas was collected with a rectal balloon using a newly developed instrument/technique, the "mucosal patch technique". The instrument allows simultaneous measurements of concentrations of granulocyte mediators in the rectal mucosa. We measured myeloperoxidase (MPO), eosinophil cationic protein (ECP), and histamine. For comparison, we made similar measurements after corn (maize) gluten challenge. RESULTS: In all coeliac patients rectal NO concentration increased after gluten challenge and reached a peak after 15 hours (mean 9464 (SEM 2393) parts per billion (ppb); range 250-24982). The maximum MPO and ECP increase occurred five hours after challenge. A correlation was found between mucosal MPO and NO production at 15 hours. Six of the patients showed an increase in NO production 15 hours after rectal corn gluten challenge but this was much smaller than after gluten challenge. No increases were seen in the control group after either challenge. CONCLUSION: Mucosal activation of neutrophils and eosinophils precedes pronounced enhancement of mucosal NO production after rectal wheat gluten challenge in patients with coeliac disease. Some of our coeliac patients displayed signs of an inflammatory reaction, as measured by NO and granulocyte markers, after rectal corn gluten challenge.
Asunto(s)
Enfermedad Celíaca/metabolismo , Glútenes/farmacología , Óxido Nítrico/metabolismo , Recto/metabolismo , Administración Rectal , Adulto , Anciano , Análisis de Varianza , Estudios de Casos y Controles , Proteína Catiónica del Eosinófilo/metabolismo , Eosinófilos/metabolismo , Femenino , Glútenes/administración & dosificación , Histamina/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Masculino , Persona de Mediana Edad , Neutrófilos/metabolismo , Peroxidasa/metabolismo , Zea maysRESUMEN
AIM: The aim of this study was to establish a method of investigating intestinal eosinophil and neutrophil granulocytes by flow cytometry, and to compare the distribution and activity of these cells in different stages of ulcerative colitis (UC). METHODS: Biopsy samples were taken from six locations of the entire colon and from the terminal ileum in 10 patients with active total UC, 10 patients with inactive total UC, eight patients with active distal UC, and 11 control subjects. Cell suspensions from biopsies and from peripheral blood were incubated with fluorophore conjugated monoclonal antibodies. The use of scatter plot-gating and specific antibodies was established in a flow cytometry assay. RESULTS: Eosinophils were more numerous and more active in patients with active UC than in controls. Interestingly, during inactive UC, the number of activated eosinophils was even larger. Eosinophil activity was high in the rectum of patients with distal colitis but was also slightly elevated in the proximal colon. Neutrophils were increased in number and activity during active but not inactive UC. In patients with distal colitis, activated neutrophils were only found in the sigmoid colon and rectum. CONCLUSION: With this method, we confirm that neutrophils participate in the inflammatory process during active UC, and that they express a resting phenotype during remission. The finding of activated eosinophils in inflamed intestine strengthens the view of these cells as proinflammatory and tissue damaging. Nevertheless, our new finding of high eosinophil activation during inactive UC suggests that eosinophils play a role in repair of injured epithelium.
Asunto(s)
Colitis Ulcerosa/patología , Eosinófilos/fisiología , Adulto , Anciano , Antígenos CD/metabolismo , Biopsia , Moléculas de Adhesión Celular/metabolismo , Células Cultivadas , Colitis Ulcerosa/tratamiento farmacológico , Eosinófilos/patología , Femenino , Citometría de Flujo/métodos , Proteínas Ligadas a GPI , Humanos , Técnicas para Inmunoenzimas , Intestino Grueso/patología , Masculino , Persona de Mediana Edad , Activación Neutrófila , Inducción de Remisión , Índice de Severidad de la EnfermedadRESUMEN
BACKGROUND AND AIMS: There is a clear need for a rapid, simple, safe, and sensitive method of determining the type and intensity of inflammation in the gut mucosa in clinical practice. In this study, we have evaluated the potential of a new method, the mucosal patch technique, in patients with and without apparent gut inflammation, as assessed by conventional diagnostic procedures. SUBJECTS AND METHODS: The technique tested is based on the idea that inflammatory mediators released from the rectal mucosa can be absorbed by and then extracted from cellulose patches brought into contact with the mucosa by use of an instrument with an inflatable balloon. Measurements were performed in healthy controls (n = 16) and in patients with active (n = 19) and inactive ulcerative colitis (UC, n = 8), collagen colitis (CC, n = 12), coeliac disease (n = 13), and irritable bowel syndrome (IBS, n = 13). RESULTS: Inflammatory mediators from neutrophils (myeloperoxidase (MPO)) and eosinophils (eosinophil cationic protein (ECP)) were increased on average 300- and 10-fold, respectively, in patients with active UC compared with healthy controls and were correlated with the endoscopic score. Patients with inactive UC, CC, coeliac disease, and IBS exhibited no endoscopic signs of inflammation. These patient groups had significantly lower levels of MPO and ECP than the active UC group but showed on average a four- to sevenfold increase in MPO compared with healthy controls. CONCLUSION: The mucosal patch technique was well tolerated by patients and easily applied by the investigator. Pronounced neutrophil and eosinophil involvement in UC was demonstrated. With the high sensitivity of the technique, low degree mucosal neutrophil activation could also be quantified in patients with CC and UC in clinical remission. The finding of increased neutrophil involvement in patients with IBS contributes to the pathophysiological ideas of this disease.
Asunto(s)
Eosinófilos/patología , Enfermedades Inflamatorias del Intestino/diagnóstico , Mucosa Intestinal/patología , Síndrome del Colon Irritable/diagnóstico , Activación Neutrófila , Adulto , Biomarcadores/análisis , Proteína Catiónica del Eosinófilo/análisis , Femenino , Humanos , Mediadores de Inflamación/metabolismo , Enfermedades Inflamatorias del Intestino/metabolismo , Enfermedades Inflamatorias del Intestino/patología , Síndrome del Colon Irritable/metabolismo , Síndrome del Colon Irritable/patología , Masculino , Persona de Mediana Edad , Cooperación del Paciente , Peroxidasa/análisis , Recto/metabolismo , Manejo de Especímenes/instrumentación , Manejo de Especímenes/métodosRESUMEN
Desmosomes are intercellular adhering junctions characteristic of epithelial cells. Several constitutive proteins--desmoplakin, plakoglobin and the transmembrane glycoproteins desmoglein and desmocollin--have been identified as fundamental constituents of desmosomes in all tissues. A number of additional and cell type-specific constituents also contribute to desmosomal plaque formation. Among these proteins is the band 6 polypeptide (B6P). This positively charged, non-glycosylated protein is a major constituent of the plaque in stratified and complex glandular epithelia. Using an overlay assay we show that purified keratins bind in vitro to B6P. Thus B6P may play a role in ordering intermediate filament networks of adjacent epithelial cells. To characterize the structure of B6P in the desmosome we have isolated cDNA clones representing the entire coding sequence. The predicted amino acid sequence of human B6P shows strong sequence homology with a murine p120 protein, which is a substrate of protein tyrosine kinase receptors and of p60v-src. P120 and B6P show amino-terminal domains differing distinctly in length and sequence. These are followed in both proteins by 460 residues that display a series of imperfect repeats corresponding to the repeats in the cadherin binding proteins armadillo, plakoglobin and beta-catenin. Over this repeat region B6P and p120 share 33% sequence identity (54% similarity). These sequence characteristics define B6P as a novel member of the armadillo multigene family and raise the question of whether the structural proteins B6P, plakoglobin, beta-catenin and armadillo share some function. Since armadillo, plakoglobin, beta-catenin and p120 seem involved in signal transduction this may also hold for B6P. The amino-terminal region of B6P (residues 1 to 263) shows no significant homology to any known protein sequence. It may therefore be involved in unique functions of B6P.
Asunto(s)
Desmosomas/genética , Proteínas de Drosophila , Familia de Multigenes/genética , Proteínas/genética , Transactivadores , Secuencia de Aminoácidos , Animales , Proteínas del Dominio Armadillo , Secuencia de Bases , Bovinos , Fraccionamiento Celular , Clonación Molecular , Proteínas del Citoesqueleto/genética , Desmocolinas , Desmogleínas , Desmoplaquinas , Desmosomas/química , Epitelio/química , Femenino , Humanos , Queratinas/metabolismo , Datos de Secuencia Molecular , Nariz/química , Placofilinas , Unión Proteica , Proteínas/metabolismo , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Transducción de Señal , beta Catenina , gamma CateninaRESUMEN
The effect of NGF on [Ca2+]i of mature regenerating oligodendroglial cells was investigated by measuring fluo-3 fluorescence. NGF caused transient increases in [Ca2+]i, which could be inhibited by anti-NGF antibody. The rise in [CA2+]i was in part due to influx of extracellular Ca2+ since it was markedly attenuated in Ca2+-free solution. It also depended on release of Ca2+ from intracellular stores as tested by prior depletion with cyclopiazonic acid. These results support a role for Ca2+ in the effects of NGF on oligodendroglial cells.
Asunto(s)
Calcio/metabolismo , Factores de Crecimiento Nervioso/farmacología , Regeneración Nerviosa/efectos de los fármacos , Oligodendroglía/efectos de los fármacos , Animales , Células Cultivadas , Senescencia Celular/fisiología , Oligodendroglía/fisiología , Transducción de Señal/efectos de los fármacos , PorcinosRESUMEN
Id-like helix-loop-helix (HLH) proteins, which lack a basic DNA binding domain, have been suggested to serve as general inhibitors of differentiation. We present data that Id2 is expressed in specific cell types during neurogenesis and in the adult. At early stages of neurogenesis, Id2 is expressed in the ventricular zone of neuroepithelium. After the first neuronal populations are born, the expression of Id2 is down regulated in neuroepithelial cells and continues to be high in Purkinje cells of the cerebellum, in mitral cells of the olfactory bulb, and in layers 2, 3, and 5 of the cerebral cortex. In neuronally differentiating cell lines, the Id2 expression is up regulated (PCC7), down regulated (NG108), or unchanged (N18) during differentiation. Analyses of promoter sequences of the Id2 gene identify the region which is responsible for the down regulation of transcription during neuronal differentiation. Our data indicate that Id2 has different functions in different cell types during neurogenesis.
Asunto(s)
Proteínas de Unión al ADN/genética , Secuencias Hélice-Asa-Hélice/genética , Sistema Nervioso/metabolismo , Neuronas/metabolismo , Proteínas Represoras , Factores de Transcripción , Animales , Secuencia de Bases , Northern Blotting , Encéfalo/embriología , Encéfalo/crecimiento & desarrollo , Diferenciación Celular , Células Cultivadas , Clonación Molecular , Cartilla de ADN , Proteínas de Unión al ADN/biosíntesis , Regulación hacia Abajo , Hibridación in Situ , Proteína 1 Inhibidora de la Diferenciación , Ratones , Datos de Secuencia Molecular , Sistema Nervioso/embriología , Sistema Nervioso/crecimiento & desarrollo , Neuronas/citología , Especificidad de Órganos/genética , Regiones Promotoras Genéticas , Mapeo RestrictivoRESUMEN
This work describes the cDNA sequence of the mouse CDEI binding protein (CDEBP), comprising the complete coding sequence. The cDNA encodes a protein of 695 amino acid residues. The derived amino acid sequence displays a sequence identity to human amyloid precursor-like protein (APLP) of > 92%.
Asunto(s)
ADN Complementario/química , Proteínas de Unión al ADN/genética , Secuencia de Aminoácidos , Precursor de Proteína beta-Amiloide/análogos & derivados , Precursor de Proteína beta-Amiloide/genética , Animales , Secuencia de Bases , Proteínas de Unión al ADN/química , Humanos , Ratones , Datos de Secuencia MolecularAsunto(s)
Comparación Transcultural , Etnicidad , Obesidad/epidemiología , Adolescente , Canadá , Niño , Femenino , Humanos , Islandia/etnología , Masculino , Factores de RiesgoRESUMEN
Antisera were raised in guinea pigs to synaptic vesicles purified from the electric organ of Torpedo marmorata. In cholinergic nerve terminals from Torpedo the major antigens identified had Mr 300,000-150,000, 86,000, and 18,000. The Mr 86,000 antigen was conserved between Torpedo and rat, where it is neuron-specific and concentrated in nerve terminals. When rat brain synaptosomes are subfractionated the antigen is associated with synaptic vesicles. The antigen is not found in the cytoskeleton and in the vesicle-free cytosol. Immunohistochemical localization of the antigen in rat shows it to be associated with synapses in diaphragm, cerebellum, hippocampus, and cerebral cortex. The staining pattern of the antigen indicates that the antigen is not cholinergic-specific. The function of the Mr 86,000 antigen remains to be identified.
Asunto(s)
Antígenos/análisis , Vesículas Sinápticas/inmunología , Torpedo/inmunología , Animales , Encéfalo/ultraestructura , Órgano Eléctrico/ultraestructura , Histocitoquímica , Técnicas Inmunológicas , Peso Molecular , Terminaciones Nerviosas/inmunología , Ratas , Sinapsis/análisis , Sinaptosomas/inmunología , Distribución TisularRESUMEN
The localization of the phosphoprotein B-50 (molecular weight 48,000 isoelectric point 4.5) in the rat has been studied. Inspection of endogenous phosphorylation patterns of the particulate as well as the cytosolic subcellular fractions from a variety of peripheral organs failed to demonstrate phosphorylation of a molecular weight 48,000 protein. Only in the particulate fractions from brain tissue was there endogenous phosphorylation of the B-50 protein. Two-dimensional analysis (isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis) and in immunochemical detection method employing an anti B-50 antiserum revealed the presence of B-50 in particulate material from brain, but not in that of other tissues. Therefore the data were interpreted as pointing to the localization of B-50 in nervous tissue. In addition, the regional distribution of endogenous B-50 phosphorylation was studied using synaptosomal plasma membranes (SPM) obtained from individual rat brain regions. The highest value was found in SPM of septal origin, the lowest in SPM from the medulla spinalis. The relationship of the high value for B-50 phosphorylation in the septum to the sensitivity of that brain area to ACTH1-24 is discussed.