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大鼠杆状环核苷酸门控阳离子通道的克隆与广泛分布

Cloning and widespread distribution of the rat rod-type cyclic nucleotide-gated cation channel.

作者信息

Ding C, Potter E D, Qiu W, Coon S L, Levine M A, Guggino S E

机构信息

Division of Endocrinology, Johns Hopkins School of Medicine, Baltimore 21205, USA.

出版信息

Am J Physiol. 1997 Apr;272(4 Pt 1):C1335-44. doi: 10.1152/ajpcell.1997.272.4.C1335.

Abstract

We used Northern blot analysis, ribonuclease protection assay (RPA), reverse transcriptase-polymerase chain reaction, and in situ hybridization to investigate the hypothesis that the CNG1 isoform of the cyclic nucleotide-gated nonselective cation channel may be widely distributed in tissues of the rat. A cDNA encoding the CNG1 isoform was isolated from rat eye and human retina, and partial sequences were isolated from rat pineal gland and human kidney. Northern blot analysis revealed a 3.1-kilobase (kb) CNG1 transcript in rat eye, pineal gland, pituitary, adrenal gland, and spleen, and a larger transcript of 3.5 kb was found in testis. RPA confirmed the identity of CNG1 mRNA in rat eye, lung, spleen, and brain. Polymerase chain reaction-based detection of the mRNA for CNG1 indicates that the channel is expressed in lower abundance in many other tissues, including thymus, skeletal muscle, heart, and parathyroid gland. The cellular distribution of CNG1 was further studied by in situ hybridization, which demonstrated expression of mRNA in lung, thymus, pineal gland, hippocampus, cerebellum, and cerebral cortex but not in heart or kidney.

摘要

我们采用Northern印迹分析、核糖核酸酶保护试验(RPA)、逆转录聚合酶链反应以及原位杂交技术,来研究环核苷酸门控非选择性阳离子通道的CNG1亚型可能在大鼠组织中广泛分布这一假说。从大鼠眼睛和人类视网膜中分离出编码CNG1亚型的cDNA,并从大鼠松果体和人类肾脏中分离出部分序列。Northern印迹分析显示,在大鼠眼睛、松果体、垂体、肾上腺和脾脏中存在一个3.1千碱基(kb)的CNG1转录本,而在睾丸中发现了一个更大的3.5 kb转录本。RPA证实了大鼠眼睛、肺、脾脏和大脑中CNG1 mRNA的一致性。基于聚合酶链反应对CNG1 mRNA的检测表明,该通道在许多其他组织中表达丰度较低,包括胸腺、骨骼肌、心脏和甲状旁腺。通过原位杂交进一步研究了CNG1的细胞分布,结果表明mRNA在肺、胸腺、松果体、海马体、小脑和大脑皮层中表达,但在心脏或肾脏中不表达。

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