鸣禽前脑基底神经节X区在鸣禽鸣唱学习中发挥重要作用，其中纹状体多棘神经元（MSN）数量最多。目前仍不清楚该类神经元同源于哺乳类背侧或腹侧纹状体神经元。Pax6与Islet-1作为高度保守的转录因子，在脊椎动物神经系统的早期发育中发挥重要作用，参与调控脑的背腹分化，分别于背侧和腹侧高表达，作为神经元背腹侧起源的标记分子。但由于这两种转录因子在细胞分化过程中表达下调，不能通过常规的免疫组化与原位杂交技术有效追踪神经元的前体细胞。Cre-loxP系统是目前追踪神经元起源的有效工具，通过构建由斑胸草雀Pax6与Islet-1基因启动子特异性驱动的Cre-loxP报告系统，可澄清MSN的胚胎期细胞起源。我们首先从ENCODE数据库获取了人Pax6与Islet-1启动子序列，然后将家鸡、斑胸草雀的基因序列与之比对，从而预测出人Pax6第四个启动子、Islet-1第三个启动子片段同为斑胸草雀Pax6与Islet-1核心启动子区，并委托公司合成了含该启动子核心片段的双荧光素酶报告基因质粒，拟验证其活性；同时，我们也证明了293T（人胚肾细胞）高表达Pax6与Islet-1，可以作为体外验证该启动子活性的有效细胞系。但目前由于疫情管控，质粒合成和运输受阻，暂未获得转染实验结果。鸣禽斑胸草雀Pax6与Islet-1基因启动子区的预测和体外验证，为后续Cre-loxP系统的建立和在体实验奠定基础。

The Area X of basal ganglia of the songbird forebrain plays an important role in songbird song learning, with the largest number of medium spiny neurons (MSN). It remains unclear that the same type of neuron originates from mammalian dorsal or ventral striatine neurons. Pax6 and Islet-1, as highly conserved transcription factors, play an important role in the early development of the vertebrate nervous system, participate in the regulation of dorsabdo-ventral differentiation of the brain, highly expressed on the dorsal and ventral sides, respectively, and act as marker molecules of neuronal dorsal and ventral origin. However, due to the failure of the expression of these two transcription factors during cell differentiation, the precursor cells of neurons cannot be effectively traced by conventional Immunohistochemical and In Situ Hybridization techniques. The Cre-loxP system is currently the most effective tool for tracing the origin of neurons, and the embryonic cell origin of MSN can be clarified by constructing a Cre-loxP reporting system driven specifically by the Promoter of the Zebra finches Pax6 and Islet-1 genes. We first obtained the human Pax6 and Islet-1 promoter sequences from the ENCODE database, and then compared the gene sequences of domestic chickens and zebra finches to predict that the fourth promoter fragment of human Pax6 and the third promoter fragment of Islet-1 are also the core promoter regions of zebra finches Pax6 and Islet-1, and commissioned the company to synthesize a double luciferase reporter plasmid containing the promoter core fragment to verify its activity. We also demonstrated that 293T (human embryonic kidney cells) with high expression of Pax6 with Islet-1 can act as an effective cell line for verifying this promoter activity in vitro. However, due to the control of the epidemic, plasmid synthesis and transportation are blocked, and the results of transfection experiments have not been obtained. Prediction and in vitro validation of the promoter regions of Songbird Zebra finches Pax6 and Islet-1 genes lays the foundation for the establishment of subsequent Cre-loxP systems and in vitro experiments.