RESUMEN
The oviduct is a dynamic organ in which final gamete maturation, fertilization and early embryo development take place. It is considered to be a sterile site; however the mechanism for sterility maintenance is still unknown. S100A7 is an anti-microbial peptide that has been reported in human reproductive tissues such as prostate, testicle, ovary, normal cervical epithelium and sperm. The current work reports the presence of S100A7 in the Fallopian tube and its localization at the apical surface of epithelial cells. For comparison, porcine S100A7 was used for antibody development and search for peptide in reproductive tissues. Although present in boar seminal vesicles and seminal plasma, S100A7 was not detected on female porcine organs. Also, in contrast with the human protein, porcine S100A7 did not show anti-microbial activity under the conditions tested. Phylogenetic analyses showed high divergence of porcine S100A7 from human, primate, bovine, ovine and equine sequences, being the murine sequence at a most distant branch. The differences in sequence homology, Escherichia coli-cidal activity, detectable presence and localization of S100A7 from human and pig, suggest that there are possible different functions in each organism.
Asunto(s)
Trompas Uterinas/metabolismo , Filogenia , Proteínas S100/metabolismo , Animales , Antibacterianos/farmacología , Bovinos , Células Epiteliales/metabolismo , Escherichia coli/efectos de los fármacos , Trompas Uterinas/citología , Femenino , Regulación de la Expresión Génica , Caballos , Masculino , Ratones , Primates , Proteína A7 de Unión a Calcio de la Familia S100 , Proteínas S100/química , Proteínas S100/genética , Proteínas S100/farmacología , Semen/metabolismo , Homología de Secuencia de Aminoácido , Ovinos , Sus scrofaRESUMEN
OBJECTIVE: We investigated the effect of advanced glycated albumin (AGE-albumin) on macrophage sensitivity to inflammation elicited by S100B calgranulin and lipopolysaccharide (LPS) and the mechanism by which HDL modulates this response. We also measured the influence of the culture medium, isolated from macrophages treated with AGE-albumin, on reverse cholesterol transport (RCT). METHODS AND RESULTS: Macrophages were incubated with control (C) or AGE-albumin in the presence or absence of HDL, followed by incubations with S100B or LPS. Also, culture medium obtained from cells treated with C- or AGE-albumin, following S100B or LPS stimulation was utilized to treat naive macrophages in order to evaluate cholesterol efflux and the expression of HDL receptors. In comparison with C-albumin, AGE-albumin, promoted a greater secretion of cytokines after stimulation with S100B or LPS. A greater amount of cytokines was also produced by macrophages treated with AGE-albumin even in the presence of HDL. Cytokine-enriched medium, drawn from incubations with AGE-albumin and S100B or LPS impaired the cholesterol efflux mediated by apoA-I (23% and 37%, respectively), HDL(2) (43% and 47%, respectively) and HDL(3) (20% and 8.5%, respectively) and reduced ABCA-1 protein level (16% and 26%, respectively). CONCLUSIONS: AGE-albumin primes macrophages for an inflammatory response impairing the RCT. Moreover, AGE-albumin abrogates the anti-inflammatory role of HDL, which may aggravate the development of atherosclerosis in DM.
Asunto(s)
Colesterol/metabolismo , Citocinas/metabolismo , Productos Finales de Glicación Avanzada/farmacología , Lipoproteínas HDL/farmacología , Macrófagos/efectos de los fármacos , Albúmina Sérica/farmacología , Transportador 1 de Casete de Unión a ATP , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 1 , Transportadoras de Casetes de Unión a ATP/metabolismo , Animales , Transporte Biológico/efectos de los fármacos , Línea Celular , Células Cultivadas , Productos Finales de Glicación Avanzada/química , Immunoblotting , Mediadores de Inflamación/metabolismo , Lipopolisacáridos/farmacología , Macrófagos/citología , Macrófagos/metabolismo , Ratones , Factores de Crecimiento Nervioso/farmacología , Subunidad beta de la Proteína de Unión al Calcio S100 , Proteínas S100/farmacología , Receptores Depuradores de Clase B/metabolismo , Albúmina Sérica/químicaRESUMEN
S100B is a soluble protein secreted by astrocytes that exerts pro-survival or pro-apoptotic effects depending on the concentration reached in the extracellular millieu. The S100B receptor termed RAGE (for receptor for advanced end glycation products) is highly expressed in the developing brain but is undetectable in normal adult brain. In this study, we show that RAGE expression is induced in cortical neurons of the ischemic penumbra. Increased RAGE expression was also observed in primary cortical neurons exposed to excitotoxic glutamate (EG). S100B exerts effects on survival pathways and neurite extension when the cortical neurons have been previously exposed to EG and these S100B effects were prevented by anti-RAGE blocking antibodies. Furthermore, nuclear factor kappa B (NF-κB) is activated by S100B in a dose- and RAGE-dependent manner and neuronal death induced by NF-κB inhibition was prevented by S100B that restored NF-κB activation levels. Together, these findings suggest that excitotoxic damage can induce RAGE expression in neurons from ischemic penumbra and demonstrate that cortical neurons respond to S100B through engagement of RAGE followed by activation of NF-κB signaling. In addition, basal NF-κB activity in neurons is crucial to modulate the extent of pro-survival or pro-death S100B effects.
Asunto(s)
Dendritas/efectos de los fármacos , Regulación de la Expresión Génica/fisiología , FN-kappa B/metabolismo , Neuronas/patología , Receptores Inmunológicos/metabolismo , Proteínas S100/farmacología , Transducción de Señal/efectos de los fármacos , Animales , Anticuerpos/farmacología , Isquemia Encefálica/patología , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Corteza Cerebral/patología , Interacciones Farmacológicas , Embrión de Mamíferos , Inhibidores Enzimáticos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Ácido Glutámico/toxicidad , Masculino , Neuronas/efectos de los fármacos , Fosfopiruvato Hidratasa/metabolismo , Ratas , Ratas Wistar , Receptor para Productos Finales de Glicación Avanzada , Receptores Inmunológicos/inmunología , Proteínas S100/metabolismo , Transducción de Señal/fisiología , Sulfadiazina/farmacología , Factores de TiempoRESUMEN
S100B is a calcium binding protein from astrocytes that regulates protein phosphorylation by binding to substrates and protein kinases. S100B might also regulate protein phosphatases and this was investigated for protein phosphatase 2B (calcineurin). The results indicate that S100B (5-10 microM) increased the activity of both purified and cytoskeletal calcineurin in a Ca-dependent manner. This effect was blocked by a specific inhibitor of calcineurin activity, but not by TRTK-12 (an inhibitor of S100B binding to other protein targets). The present results and the known co-localization of S100B and calcineurin in the astrocyte cytoskeleton suggest that S100B may play a role in the phosphorylation state of cytoskeletal proteins.
Asunto(s)
Astrocitos/enzimología , Calcineurina/metabolismo , Proteínas del Citoesqueleto/metabolismo , Factores de Crecimiento Nervioso/metabolismo , Proteínas S100/metabolismo , Animales , Animales Recién Nacidos , Astrocitos/efectos de los fármacos , Astrocitos/ultraestructura , Calcineurina/efectos de los fármacos , Calcio/metabolismo , Señalización del Calcio/efectos de los fármacos , Señalización del Calcio/fisiología , Células Cultivadas , Proteínas del Citoesqueleto/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Factores de Crecimiento Nervioso/farmacología , Fosforilación/efectos de los fármacos , Ratas , Ratas Wistar , Subunidad beta de la Proteína de Unión al Calcio S100 , Proteínas S100/farmacología , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/fisiologíaRESUMEN
1. Stellation of astrocytes in culture involves a complex rearrangement of microfilaments, intermediate filaments, and microtubules, which reflects in part the plasticity of these cells observed during development or after injury. 2. An astrocytic calcium-binding protein, S100B, has been implicated in the regulation of plasticity due to its ability to interact with cytoskeletal proteins. 3. We used digitonin-permeabilized astrocytes to introduce TRTK-12, a peptide that binds to the C-terminal of S100B and blocks its interaction with cytoskeletal proteins. 4. TRTK-12 was able to block cAMP-induced astrocyte stellation and this effect was dependent on the concentration of the peptide. These results support the idea that S100B has a modulatory role on astrocyte morphology.
Asunto(s)
Astrocitos/metabolismo , AMP Cíclico/metabolismo , Citoesqueleto/metabolismo , Digitonina/metabolismo , Factores de Crecimiento Nervioso/metabolismo , Oligopéptidos/metabolismo , Proteínas S100/metabolismo , Animales , Astrocitos/efectos de los fármacos , Proteína CapZ , Células Cultivadas , AMP Cíclico/farmacología , Citoesqueleto/efectos de los fármacos , Digitonina/farmacología , Relación Dosis-Respuesta a Droga , Factores de Crecimiento Nervioso/farmacología , Oligopéptidos/farmacología , Fragmentos de Péptidos , Permeabilidad/efectos de los fármacos , Ratas , Ratas Wistar , Subunidad beta de la Proteína de Unión al Calcio S100 , Proteínas S100/farmacologíaRESUMEN
Adult male Wistar rats were bilaterally implanted with indwelling cannulae in the hippocampus. Forty-eight hours after surgery, animals were habituated to an open-field box during 2 min, being tested 24 h later; next they were trained in a step-down inhibitory avoidance task (3.0 s, 0.4 mA foot-shock), being tested again 24 h later. Immediately after the training session of each task, animals received a 0.5-microl infusion of calcium-phosphate-buffered saline (PBS) and S100B (20, 200, 2000, or 20,000 nM). In the inhibitory avoidance task, animals infused with the two highest concentrations of S100B, 2 and 20 microM, obtained higher scores of retention relative to controls in the test session (p<0.05), and a trend toward an increase was observed in animals infused with 200 nM (p<0. 10). In both sessions of the habituation task, groups were not different regarding crossings, rearings, and time for leaving the first square (p>0.10). These results indicate that, in rats, post-training increased hippocampal levels of S100B right after training facilitate, in a dose-dependent way, long-term memory for an inhibitory avoidance task, but not for an open-field habituation.