论文部分内容阅读
目的环核苷酸磷酸二酯酶(phosphodiesterase,PDE)是一氧化氮(nitric oxide,NO)信号转导途径中的一个关键成员,在认知与学习、帕金森病、注意缺陷多动障碍、精神病和抑郁症等过程中发挥重要作用。PDE在豚鼠脑中的表达情况还未见报道。本研究旨在对豚鼠脑中表达的Pde cDNA序列进行鉴定与分析。方法根据人PDE和小鼠Pde cDNA序列的22个保守区设计引物,采用RT-PCR法扩增豚鼠脑中表达的未知Pde基因cDNA部分序列,并采用生物信息学软件分析不同物种间Pde cDNA序列及氨基酸多肽序列的同源性及保守功能基序。结果在豚鼠脑中发现了11个新的Pde基因cDNAs,包括CpPde1a、CpPde1b、CpPde2a、CpPde4a、CpPde4d、CpPde5a、CpPde6c、CpPde7b、CpPde8a、CpPde9a和CpPde10a。豚鼠与人之间的Pde基因cDNA序列的同源率在83.8%-94.3%之间,氨基酸序列的同源率在91.9%-100%之间。豚鼠与小鼠之间的Pde基因cDNA序列的同源率在84.6%-92.1%之间,氨基酸序列的同源率在91.2%-99.2%之间。此外,这11个Pde cDNAs在豚鼠与人之间的平均同源率显著高于在豚鼠与小鼠之间的同源率(P<0.01)。豚鼠的部分氨基酸序列具有至少一个人及小鼠PDE蛋白的保守功能区。结论这些发现为进一步研究PDE在大脑中的生理功能打下了基础。
The target cyclic nucleotide phosphodiesterase (PDE) is a key member of the nitric oxide (NO) signal transduction pathway, which is involved in cognition and learning, Parkinson’s disease, attention deficit hyperactivity disorder, Mental illness and depression play an important role in the process. PDE expression in guinea pig brain has not been reported. The aim of this study was to identify and analyze the Pde cDNA sequence expressed in guinea pig brains. Methods Based on the 22 conserved regions of human PDE and mouse Pde cDNA sequences, primers were designed and the cDNA sequence of unknown Pde gene was amplified by RT-PCR. The Pde cDNA sequence of different species was analyzed by bioinformatics software And amino acid polypeptide sequence homology and conservative function motifs. Results Eleven new Pde gene cDNAs were found in guinea pig brains including CpPde1a, CpPde1b, CpPde2a, CpPde4a, CpPde4d, CpPde5a, CpPde6c, CpPde7b, CpPde8a, CpPde9a and CpPde10a. The homology of the Pde gene cDNA sequence between guinea pigs and human was between 83.8% -94.3%, and the amino acid sequence homology was between 91.9% -100%. The homology of the Pde gene cDNA sequences between guinea pigs and mice ranged from 84.6% to 92.1%, and the amino acid sequence identities ranged from 91.2% to 99.2%. In addition, the average homology between these 12 Pde cDNAs in guinea pigs and humans was significantly higher than that between guinea pigs and mice (P <0.01). The partial amino acid sequence of a guinea pig has a conserved functional domain of at least one human and mouse PDE protein. Conclusions These findings provide the basis for further studies on the physiological functions of PDE in the brain.