Piketty V, Pelletier J
Laboratoire de Neuroendocrinologie Sexuelle, INRA, Nouzilly, France.
Neuroendocrinology. 1993 Sep;58(3):359-65. doi: 10.1159/000126563.
After validation of the methodology, melatonin receptors have been measured by binding of (125I)-melatonin to membranes of individual pars tuberalis/median eminence of lambs at different times of the day in the course of three experiments. Plasma melatonin was assessed by radioimmunoassay 60 min, 30 min and just before slaughter. Kd was found in the range of 10-25 pM and did not vary with the time of slaughter. Particularly, the mean Kd values were identical and equal to 17 pM in animals slaughtered either during the light or the dark phase of the day. The pattern of Bmax changes was similar in the three experiments and varied significantly (p < 0.001) with the time of the day. The apparent numbers of receptors were found to be the highest at the end of the day and at the onset of the night and to be the lowest at the end of the night: 63.4 +/- 6.3, 61.4 +/- 6.5 and 34.4 +/- 2.8 fmol/mg protein (mean +/- SEM), respectively. Values were found to be intermediary either at the middle of the day (51.1 +/- 5.3 fmol/mg protein) or at the middle of the night (40.4 +/- 6.3 fmol/mg protein). Furthermore, plasma melatonin was positively correlated to the melatonin receptor number (p < 0.02) but only at the onset of the night. In conclusion, the results are strongly suggestive of the existence of a circadian rhythm in the apparent number of melatonin receptors in the lamb pars tuberalis/median eminence with a possible downregulation occurring during the night.
在该方法得到验证后,在三个实验过程中,于一天中的不同时间,通过(125I)-褪黑素与羔羊个体结节部/正中隆起膜的结合来测定褪黑素受体。在屠宰前60分钟、30分钟以及即将屠宰时,通过放射免疫分析法评估血浆褪黑素水平。发现解离常数(Kd)在10 - 25皮摩尔范围内,且不随屠宰时间而变化。具体而言,在白天的光照期或黑暗期屠宰的动物中,平均Kd值相同,均为17皮摩尔。在这三个实验中,最大结合容量(Bmax)的变化模式相似,且随一天中的时间显著变化(p < 0.001)。发现受体的表观数量在一天结束时和夜晚开始时最高,而在夜晚结束时最低:分别为63.4 +/- 6.3、61.4 +/- 6.5和34.4 +/- 2.8飞摩尔/毫克蛋白质(平均值 +/- 标准误)。在白天中间时段(51.1 +/- 5.3飞摩尔/毫克蛋白质)或午夜时段(40.4 +/- 6.3飞摩尔/毫克蛋白质)的值处于中间水平。此外,血浆褪黑素仅在夜晚开始时与褪黑素受体数量呈正相关(p < 0.02)。总之,结果强烈提示羔羊结节部/正中隆起中褪黑素受体的表观数量存在昼夜节律,且可能在夜间发生下调。