RESUMEN
The aim of this study was to identify molecular pathways involved in audiogenic seizures in the epilepsy-prone Wistar Audiogenic Rat (WAR). For this, we used a suppression-subtractive hybridization (SSH) library from the hippocampus of WARs coupled to microarray comparative gene expression analysis, followed by Northern blot validation of individual genes. We discovered that the levels of the non-protein coding (npc) RNA BC1 were significantly reduced in the hippocampus of WARs submitted to repeated audiogenic seizures (audiogenic kindling) when compared to Wistar resistant rats and to both naive WARs and Wistars. By quantitative in situ hybridization, we verified lower levels of BC1 RNA in the GD-hilus and significant signal ratio reduction in the stratum radiatum and stratum pyramidale of hippocampal CA3 subfield of audiogenic kindled animals. Functional results recently obtained in a BC1â»/â» mouse model and our current data are supportive of a potential disruption in signaling pathways, upstream of BC1, associated with the seizure susceptibility of WARs.
Asunto(s)
Regulación hacia Abajo/genética , Epilepsia Refleja/patología , Hipocampo/metabolismo , Excitación Neurológica/genética , ARN Citoplasmático Pequeño/genética , ARN no Traducido/genética , Estimulación Acústica/efectos adversos , Animales , Modelos Animales de Enfermedad , Electroencefalografía , Epilepsia Refleja/genética , Perfilación de la Expresión Génica/métodos , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , ARN Citoplasmático Pequeño/metabolismo , Ratas , Ratas WistarRESUMEN
OBJECTIVE: To analyze the protein and messenger RNA (mRNA) expression of La/SSB, Ro/SSA 60, and Ro/SSA 52 antigens in minor salivary glands (MSG) from patients with primary Sjögren's syndrome (pSS). METHODS: La/SSB, Ro/SSA 60, and Ro/SSA 52 protein expression was studied by immunohistochemistry in MSG from 26 patients with pSS and 16 controls. mRNA expression was determined by real-time polymerase chain reaction in MSG of 10 patients with pSS and 7 controls. RESULTS: La/SSB and Ro/SSA 60, but not Ro/SSA 52, mRNA expression was higher in samples from patients with pSS compared to controls (p < 0.05). La/SSB protein had higher expression in the cytoplasm of ductal cells than in the cytoplasm of mucous acinar cells in patients with pSS (p = 0.013) but not in controls. Ro/SSA 60 had higher expression in the cytoplasm of ductal cells than in the cytoplasm of serous acinar cells in patients with pSS (p = 0.006) but not in controls. The Ro/SSA 52 expression pattern was similar in patients and controls. There was no association between circulating autoantibodies to Ro/SSA or La/SSB and the aberrant expression of the cognate autoantigens. CONCLUSION: The increased Ro/SSA 60 and La/SSB mRNA expression in MSG of patients with pSS as well as the differential Ro/SSA 60 and La/SSB protein expression in ductal cells of MSG in patients with pSS suggest that these these 2 autoantigens, but not Ro/SSA 52, are probably involved in triggering and maintaining the tissue-specific autoimmune response in pSS MSG and may contribute to the antigen-driven immune response and local autoantibody production in pSS.