RESUMO
OBJECTIVE: To develop radioimmunoassay for hypocretin-2 (Hcrt-2). And search for its presence in certain rat tissues. METHODS: Anti-Hcrt-2 serum has been raised in New Zealand white rabbits immunized with a conjugate of synthetic Hcrt-2 with bovine serum albumin. Radioiodination of Hcrt-2 was performed by chloramine T method, followed by purification of radoiodinated material on Sephadex G-25 column. RESULTS: The obtained antibody did not cross react with hypocretin-2, hypothalamic hormones, pituitary hormones, neuropeptides or gut hormones. The assay was performed with a double antibody system. Hcrt-2 was extracted from the tissues with acid acetone. The dilution curve of acid acetone extracts of rat hypothalamus in the radioimmunoassay system was parallel to the standard curve. The recovery of tissue Hcrt-2 was about 85 % and the intra-assay and inter-assay variation were 5.6 % and 8.0 %, respectively. Hcrt-2 was found in the hypothalamus, cerebrum, brain stem and testes. CONCLUSIONS: The obtained data suggest that the assay system developed is suitable to measure Hcrt-2 in tissues and that Hcrt-2 is mainly found in the hypothalamus.
Assuntos
Neuropeptídeos , Neurotransmissores/análise , Radioimunoensaio , Animais , Anticorpos , Especificidade de Anticorpos , Química Encefálica , Tronco Encefálico/química , Hipotálamo/química , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Neurotransmissores/imunologia , Orexinas , Especificidade de Órgãos , Controle de Qualidade , Coelhos , Ratos , Ratos Wistar , Testículo/químicaRESUMO
Subacute toxicity of solvent adjuvant, alphadolone acetate and solvent, 20 per cent Cremophor solution contained in CT-1341 was studied by using rats of both sexes. Alphadolone acetate and Cremophor solution were intraperitoneally injected every day for a period of one month. Total rats tolerated to daily administration of 60 mg/kg of alphadolone acetate or of 20 ml/kg of 20 per cent Cremophor solution, without showing significant changes in body weight curves and food intake. No change was observed in blood cells and in biochemical data of blood and urine as compared with control. Rats subjected to daily administration of 60 mg/kg of alphadolone acetate presented slight patho-histological changes such as swelling of cells of the liver and kidneys, and also cell infiltration of pericapillary tissues of the lung.