Song Y, Pang C S, Ayre E A, Brown G M, Pang S F
Clarke Institute of Psychiatry, Toronto, Ontario, Canada.
Eur J Endocrinol. 1996 Jul;135(1):128-33. doi: 10.1530/eje.0.1350128.
The effect of pinealectomy on the characteristics of melatonin receptors in the chicken kidney was studied. One-day-old chicks were operated and kept under a 12 h/12 h light/dark photoperiod. Six weeks after operation, the animals were sacrificed at mid-light and mid-dark. Serum melatonin was determined by radioimmunoassay and kidney melatonin receptors were studied by radioreceptor assay using the melatonin agonist 2-[125I]iodomelatonin as the radioligand. Pinealectomy significantly reduced the mid-dark serum melatonin level and abolished the diurnal rhythm of 2-[125I]-iodomelatonin binding in the kidney. The density of 2-[125I]-Iodomelatonin binding sites in the kidney at mid-dark was increased significantly to a value comparable to the mid-light density after pineal ablation. Our results suggest that melatonin receptors in the chicken kidney are directly regulated by melatonin in the circulation. The coupling of kidney melatonin receptors to adenylate cyclase was investigated. The basal and forskolin-stimulated cAMP production in chicken kidney explants was studied following melatonin or melatonin plus pertussis toxin treatment. Levels of cAMP in chicken kidney explants were extracted and determined by radioimmunoassay. Melatonin had no effect on basal cAMP levels. However, melatonin significantly inhibited the forskolin-stimulated cAMP accumulation at a concentration of 10 pmol/l. Inhibitory effects of melatonin on the forskolin-stimulated cAMP increase in the chicken kidney were totally blocked by preincubating the kidney tissue with 1.0 micrograms/ml pertussis toxin. Our results suggest that kidney melatonin receptors may modulate the adenylate cyclase leading to biological responses in the renal system.
研究了松果体切除对鸡肾脏褪黑素受体特性的影响。将1日龄雏鸡进行手术,并饲养在12小时光照/12小时黑暗的光周期下。术后6周,在光照中期和黑暗中期处死动物。通过放射免疫测定法测定血清褪黑素,并使用褪黑素激动剂2-[125I]碘褪黑素作为放射性配体,通过放射受体测定法研究肾脏褪黑素受体。松果体切除显著降低了黑暗中期的血清褪黑素水平,并消除了肾脏中2-[125I]碘褪黑素结合的昼夜节律。松果体切除后,黑暗中期肾脏中2-[125I]碘褪黑素结合位点的密度显著增加,达到与光照中期密度相当的值。我们的结果表明,鸡肾脏中的褪黑素受体直接受循环中褪黑素的调节。研究了肾脏褪黑素受体与腺苷酸环化酶的偶联。在褪黑素或褪黑素加百日咳毒素处理后,研究了鸡肾脏外植体中基础和福司可林刺激的cAMP产生。提取鸡肾脏外植体中的cAMP水平,并通过放射免疫测定法测定。褪黑素对基础cAMP水平没有影响。然而,褪黑素在浓度为10 pmol/l时显著抑制了福司可林刺激的cAMP积累。用1.0微克/毫升百日咳毒素预孵育肾脏组织可完全阻断褪黑素对鸡肾脏中福司可林刺激的cAMP增加的抑制作用。我们的结果表明,肾脏褪黑素受体可能调节腺苷酸环化酶,从而导致肾脏系统中的生物学反应。