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犬脑钠肽的互补DNA克隆、组织分布及合成

Complementary DNA cloning, tissue distribution, and synthesis of canine brain natriuretic peptide.

作者信息

Asano K, Murakami M, Endo D, Kimura T, Fujinaga T

机构信息

Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.

出版信息

Am J Vet Res. 1999 Jul;60(7):860-4.

Abstract

OBJECTIVE

To determine complimentary DNA (cDNA) sequence and tissue distribution of canine brain natriuretic peptide (BNP), and to investigate whether synthesis of canine BNP increases in association with cardiovascular dysfunction.

ANIMALS

5 healthy adult mixed-breed dogs and 3 healthy adult Beagles.

PROCEDURE

Total RNA was extracted from normal canine hearts and was used in a reverse transcription-polymerase chain reaction (RT-PCR) procedure to isolate canine BNP cDNA. Sequence of the isolated cDNA was analyzed. Gene expression of canine BNP in various tissues from 2 mixed-breed dogs was investigated, using RT-PCR and northern blot analyses. Moreover, messenger RNA (mRNA) expression of canine BNP, using northern blot analysis, was compared between grossly normal hearts from 3 Beagles and hearts from 3 mixed-breed dogs with acute myocardial infarction created by surgical ligation.

RESULTS

The cDNA sequence and deduced amino acid residues of canine BNP precursor were 420 base pairs and 140 residues, respectively. Messenger RNA expression of canine BNP was detectable in the atria but not in the ventricles and the other tissues. Messenger RNA expression of canine BNP was, however, detectable in the infarcted portion of the ventricles. The amount of canine BNP mRNA in the infarcted ventricles was significantly increased, compared with that of noninfarcted ventricles.

CONCLUSION

The cDNA sequence of canine BNP was determined. Expression of canine BNP mRNA was detected not only in the atria but also in infarcted ventricles. Synthesis of canine BNP increases in association with ischemic myocardial injury. Canine BNP may be used as an indicator of severity of ventricular myocardial injury.

摘要

目的

确定犬脑钠肽(BNP)的互补DNA(cDNA)序列和组织分布,并研究犬BNP的合成是否随心血管功能障碍而增加。

动物

5只健康成年杂种犬和3只健康成年比格犬。

步骤

从正常犬心脏中提取总RNA,并用于逆转录-聚合酶链反应(RT-PCR)程序以分离犬BNP cDNA。分析分离出的cDNA序列。使用RT-PCR和Northern印迹分析研究2只杂种犬不同组织中犬BNP的基因表达。此外,使用Northern印迹分析比较了3只比格犬大体正常心脏和3只通过手术结扎造成急性心肌梗死的杂种犬心脏中犬BNP的信使核糖核酸(mRNA)表达。

结果

犬BNP前体的cDNA序列和推导的氨基酸残基分别为420个碱基对和140个残基。犬BNP的mRNA表达在心房中可检测到,但在心室和其他组织中未检测到。然而,在心室梗死部位可检测到犬BNP的mRNA表达。与未梗死心室相比,梗死心室中犬BNP mRNA的量显著增加。

结论

确定了犬BNP的cDNA序列。犬BNP mRNA不仅在心房中表达,在梗死心室中也有表达。犬BNP的合成随缺血性心肌损伤而增加。犬BNP可作为心室心肌损伤严重程度的指标。

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