Abstract： Layer pinealocytes were isolated by using double digestion methods of collagenase and trypsin, and cells were divided into 3 groups, which were the control （medium）, alcohol（ solvent control）, and MT （mixed with alcohol） group. In order to observe the effects of Melatonin（MT） on the expression of GnRH - I gene in layers pinealocytes cultured in vitro, total RNA was extracted from cells on 11 th day of culture, and relative RT -PCR method was employed to detect GnRH -I mRNA expression using β -actin as the internal control. The results showed that the MTT value of pinealocytes increased progressively from 1 to 9 days of culture in vitro, andthere was a bit but not significant decrease on the 11 th day of culture compared to that of the 9th day. The expression abundance of GnRH - I mRNA in MT group were significantly higher than that of the control group （ P 〈 0.05 ）, and still has up - regulated the expression trend by MT treatment for seven days compared with alcohol group（P 〈0.1 ）, while there was no significant difference between control and alcohol groups （P 〉 0.05 ）. These results indicated that GnRH - I gene expression in layer pinealocytes cultured in vitro was significantly up regulated by exogenous MT treatment.%通过胶原酶、胰蛋白酶双消化法获取蛋鸡松果体细胞，分为对照（培养基）、酒精（溶剂对照）和褪黑索（MT）三组进行原代细胞培养，采用相对定量RT-PCR方法检测褪黑素（MT）对培养的松果体细胞促性腺激素释放激素-I（GnRH-I）基因表达的影响。结果表明：MTr检测正：常培养的松果体细胞增殖速度第9d达高峰，11d缓降。与对照组比较，MT处理1周显著上调其表达丰度（P〈0．05）；与酒精溶剂组相比。MT处理1周具有上调其表达的趋势（P〈0．1）；对照组与酒精组之间没有明显的差异（P〉0．05）。从而提示：体外培养松果体细胞的GnRH表达受MT的调控。
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