Biondo Alexander W, Wiedmeyer Charles E, Sisson David D, Solter Philip F
Department of Veterinary Pathobiology, University of Illinois at Urbana-Champaign, Urbana, IL 61802, USA.
Vet Clin Pathol. 2003;32(4):188-94. doi: 10.1111/j.1939-165x.2003.tb00334.x.
Endothelin-1 (ET-1, "mature ET-1") is a potent vasoconstrictor peptide that is made along with "big ET-1" from its precursor, preproET-1. Increased plasma concentrations of ET-1 and big ET-1 occur with various forms of cardiovascular disease in humans. Our laboratory is investigating plasma endothelins as diagnostic tests of cardiovascular disease in dogs and cats; however, commercial immunoassays designed specifically for use in dogs and cats are limited.
Amino acid sequences of feline and canine big ET-1 were obtained and used to predict antibody cross-reactivity with immunoassay test kits from other species.
Genomic DNA was extracted from peripheral blood and total RNA was extracted from canine and feline left ventricles for reverse transcription polymerase chain reaction (PCR) and PCR amplification of segments of the canine and feline preprohormone containing the big ET-1 sequences. The derived amino acid sequences were compared with known big ET-1 and ET-1 sequences of several other species, including human, mouse, and rat.
Feline and canine big ET-1 had 87-97% and 89-100% homology, respectively, with that of other mammalian species. Canine ET-1 was identical to human, mouse, and rat ET-1. In contrast, the amino acid sequence of feline ET-1 was unique owing to a leucine for methionine substitution at position 7.
It is highly likely that anti-human and anti-rodent ET-1 antibodies will cross-react with mature canine ET-1. In contrast, antibodies to mature ET-1 intended for use with feline tissues and antibodies to big ET-1 in either dogs or cats may have partial or no cross-reactivity depending on the peptide sequences used to produce the antibodies.
内皮素-1（ET-1，“成熟ET-1”）是一种强效血管收缩肽，它与其前体前内皮素原-1一起产生“大ET-1”。人类各种形式的心血管疾病会导致血浆中ET-1和大ET-1浓度升高。我们实验室正在研究血浆内皮素作为犬猫心血管疾病的诊断测试；然而，专门用于犬猫的商业免疫测定法有限。
获得猫和犬大ET-1的氨基酸序列，并用于预测与其他物种免疫测定试剂盒的抗体交叉反应性。
从外周血中提取基因组DNA，从犬猫左心室中提取总RNA，用于逆转录聚合酶链反应（PCR）以及对包含大ET-1序列的犬猫前激素原片段进行PCR扩增。将推导的氨基酸序列与包括人、小鼠和大鼠在内的其他几种物种的已知大ET-1和ET-1序列进行比较。
猫和犬大ET-1与其他哺乳动物物种的同源性分别为87 - 97%和89 - 100%。犬ET-1与人、小鼠和大鼠的ET-1相同。相比之下，猫ET-1的氨基酸序列是独特的，因为在第7位有亮氨酸取代甲硫氨酸。
抗人及抗啮齿动物ET-1抗体极有可能与成熟犬ET-1发生交叉反应。相比之下，用于猫组织的成熟ET-1抗体以及犬或猫中大ET-1的抗体可能会有部分交叉反应或无交叉反应，这取决于用于产生抗体的肽序列。
