Modulation of 2[ 125I]iodomelatonin binding sites in the guinea pig spleen by melatonin injection is dependent on the dose and period but not the time

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Effects of dose, time and period of melatonin injection on 2[ 125I]iodomelatonin binding sites in the guinea pig spleen were studied. Guinea pigs ( Dunkin-hartley), kept under 12h light/12h darkness, were given daily intraperitoneal injections of either vehicle or 0.01, 0.1 or 1 mg melatonin/kg body weight in either early (1 hour after of light period) or late light period (1 hour before offset of light period) for 2 or 7 days. To study the effect of opioid antagonist on the binding, intraperitoneal injections of 2 or 20 mg naltrexone/kg body weight alone or together with 0.1 mg melatonin/kg body weight was given daily in late light period for 2 days. 2[ 125I]Iodomelatonin binding assays were performed on spleen membrane preparations and radioimminoassays of melatonin levels were carried out in serum and pineal glands collected during mid-light. High dose (1 mg/kg body weight) of melatonin injection elevated the mid-light serum melatonin levels without affecting pineal melatonin levels. Early light injection group had a higher mid-light serum melatonin level. Melatonin injection for 2 days at either time points caused a dose-dependent decrease in Bmax and increase in Kd of 2[ 125I]iodomelatonin binding sites in the spleen. The response was independent of the time of injection. A greater suppression of binding was achieved by injecting melatonin for 7 days. Naltrexone did not affect the binding by itself and was not able to reverse the melatonin-induced suppression of binding in the spleen. The modulation of the splenic 2[ 125I]iodomelatonin binding sites by exogenous melatonin suggests that melatonin may act directly on the immune system to affect its function.