Identification of novel cyclic nucleotide phosphodiesterase gene cDNAs in the brain of guinea pig

Abstract

Objective

Cyclic nucleotide phosphodiesterase (PDE) is a critical component of the nitric oxide (NO) signaling pathway and plays critical roles in cognition and learning, Parkinson’s disease, attention deficit hyperactivity disorder, psychosis and depression. The PDEs in the brain of guinea pig have not yet been reported. The present study aimed to detect the unknown Pde cDNAs in the brain of guinea pig.

Methods

Reverse transcription polymerase chain reaction (RT-PCR) and sequence comparison analysis were performed to detect the expression of Pde cDNAs and to assess the identity rates of cDNA and amino acid sequences between guinea pig and human or mouse, respectvely. The RT-PCR primers were located on the conserved region of human PDE and mouse Pde cDNAs.

Results

Eleven novel Pde cDNAs were detected in the brain of guinea pig (Cavia porcellus), including CpPde1a, CpPde1b, CpPde2a, CpPde4a, CpPde4d, CpPde5a, CpPde6c, CpPde7b, CpPde8a, CpPde9a, and CpPde10a. The identity rates of the Pde cDNA sequences between guinea pig and human ranged from 83.8% to 94.3%, and those of the amino acid sequences ranged from 91.9% to 100%. The identity rates of Pde cDNA sequences between guinea pig and mouse ranged from 84.6% to 92.1%, and those of amino acid sequences ranged from 91.2% to 99.2%. The average identity rate of the 11 Pde cDNA sequences between guinea pig and human was significantly higher (P < 0.01) than that between guinea pig and mouse. The putative partial amino acid sequences of guinea pig contained at least one of the conserved domains of human and mouse PDE proteins.

Conclusion

These results indicate that the brainexpressed Pde genes are identified in guinea pig, which lays the foundation for further investigating the physiological roles of PDE proteins in the brain.

摘要

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。