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1.
Invest. clín ; 54(3): 257-269, sep. 2013. ilus
Article in English | LILACS | ID: lil-740324

ABSTRACT

Neuregulins (NRG) are proteins that belong to the family of epidermal growth factors. It is well established that these factors are essential for the development and maintenance of the nervous system. Due to the difficulty of purifying enough quantities of these factors and the lack of specificity from commercially available antibodies, the aim of this work was to produce antibodies against a synthetic peptide capable to detect and identify neuregulin GGFb isoforms. To accomplish this goal, polyclonal antibodies were raised in hens against a synthetic peptide designed from the GGFb1 extracellular sequence. The sequence analysis was made using different epitope-predicting programs. Our results showed that the peptide sequence selected was immunogenic because it was capable of inducing a specific type B immune response in the experimental animal model. These antibodies were also capable of recognizing a recombinant GGF protein and GGF isoforms present in different samples. Our results suggest that the development of immunoglobulin Y (IgY) using synthetic peptides represents, a valuable tool for neuroscience research.


Las Neuregulinas (NRG) son proteínas que pertenecen a la familia de los factores de crecimiento epidermal. Se ha demostrado que estos factores son esenciales para el desarrollo y mantenimiento de la funcionalidad del sistema nervioso. Debido a la dificultad para purificar estas proteínas y la falta de especificidad de los anticuerpos disponibles comercialmente, el objetivo de este trabajo fue producir anticuerpos contra un péptido sintético capaz de detectar e identificar una isoforma de la Neuregulina (GGFb). Para lograr este objetivo, se desarrollaron anticuerpos en gallinas (IgY) contra un péptido sintético diseñado a partir de la secuencia aminoacídica de la región extracelular de GGFb, utilizando programas de predicción de epítopes. Los resultados demuestran que el péptido seleccionado fue immunogénico debido a que estimuló una respuesta inmune específica tipo B en el modelo utilizado. Estos anticuerpos fueron también capaces de reconocer una proteína recombinante e isoformas de GGF presentes en diferentes muestras biológicas. Nuestros resultados demuestran el potencial valor de las inmunoglobulinas Y (IgY) contra péptidos sintéticos como una herramienta de aplicación para la investigación en neurociencia.


Subject(s)
Animals , Female , Rats , Antibodies, Heterophile/immunology , Chickens/immunology , Immunoglobulins/immunology , Neuregulin-1/immunology , Peptide Fragments/immunology , Antibody Specificity , Antibodies, Heterophile/biosynthesis , Antibodies, Heterophile/isolation & purification , Cells, Cultured , Culture Media, Conditioned , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , Epitopes/immunology , Immunoblotting , Immunoglobulins/biosynthesis , Immunoglobulins/isolation & purification , Neuregulin-1/analysis , Peptide Fragments/chemical synthesis , Protein Isoforms/analysis , Protein Isoforms/immunology , Rats, Sprague-Dawley , Recombinant Proteins/immunology , Schwann Cells/immunology , Schwann Cells/metabolism , Sciatic Nerve/cytology
2.
Chinese Journal of Endocrine Surgery ; (6): 230-234, 2011.
Article in Chinese | WPRIM | ID: wpr-622245

ABSTRACT

ObjectiveTo explore the relationship between glial growth factor (GGF), nerve growth factor (NGF) and growth characteristics of prolactinoma (PRL) and to evaluate pre and postoperative growth of PRL. MethodsImmunohistochemistry was used to analyze expression of GGF and NGF in 86 cases of PRL and to analyze the relationship between expression of GGF, NGF and PRL level, invasion, stroke, microvessel density. Cells were cultured with GGF and NGF to observe cell growth, cell cycle and angiogenesis. The relationship between proliferation, growth rate and GGF, NGF was evaluated by rank correlation and Chi-square test. Results GGF expression was significantly higher in invasive, stroke and recurrent pituitary adenomas ( P < 0.05 ).Microvessel density increased significantly ( P < 0.01 ). NGF expression was significantly lower in invasive, apoplexy and recurrent adenomas ( P < 0. 05 ). Microvessel density decreased dramatically with NGF interruption ( P < 0.05 ). GGF showed a positive correlation with growth rate of PRL. NGF showed a negative correlation with invasion and stroke. ConclusionsGGF is one of the factors facilitating growth and invasion of PRL while NGF can partly restrain proliferation of PRL cells. Expression of GGF and NGF indirectly reflects proliferation activity of PRL and can be used as markers to evaluate invasion, recurrence, treatment response and prognosis of PRL.

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