珍贵绢丝昆虫天蚕FXPRLamide家族神经肽的免疫组织化学鉴定.pdfVIP

中国农业科学 2005,38(12):2542-2548 Scientia Agricultura Sinica 珍贵绢丝昆虫天蚕FXPRLamide家族神经肽的 免疫组织化学鉴定 1 2 魏兆军 ，赵 远 1 2 （合肥工业大学生物工程系，合肥 230009 ； 中国农业科学院蚕业研究所，镇江 212018 ） 摘要：利用Zooblotting、酶联免疫吸附测定（ELISA）和免疫组织化学等方法，对天蚕的DH、PBAN等FXPRLamide 家族神经肽进行鉴定。结果表明，标记的蓖麻蚕的DH-PBAN cDNA片断能够与天蚕、樟蚕、柳蚕和棉铃虫的基因组 DNA 结合。这证明了天蚕可能存在 FXPRLamide 家族神经肽中的 DH－PBAN 基因；天蚕的咽下神经节中 FXPRLamide 神经肽含量最高，咽下神经节有三簇细胞合成FXPRLamide神经肽，在胸神经节和腹神经节也有明显的阳性细胞； 天蚕5龄幼虫的血淋巴中FXPRLamide神经肽含量比较稳定，化蛹前含量升高，蛹期含量降低。上述结果证明天蚕 含有DH、PBAN 等FXPRLamide 家族神经肽，其对天蚕的滞育和性信息素合成等的调控还需要进一步研究。 关键词：天蚕；FXPRLamide家族神经肽；免疫组织化学；鉴定 Identification of FXPRLamide Family Neuropeptides from the Japanese Oak Silkworm, Antheraea yamamai by Using Immunocytochemistry Methods 1 2 WEI Zhao-jun , ZHAO Yuan 1 2 ( Department of Biotechnology, Hefei University of Technology, Hefei 230009; The Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang 212018) Abstract: In present study, zooblooting, ELISA and whole-mount immunocytochemistry methods were used to identify the FXPRLamide family neuropeptides from the Japanese oak silkworm, Antheraea yamamai. The results showed that the genomic DNA from Antheraea yamamai showed positive bands after hybridized with the fragment of DH-PBAN cDNA from Samia cynthia ricini, which labeled with [α-32P]-dCTP. The SG showed highest FXPRLamide peptides titer in neural o