Dackor Ryan T, Fritz-Six Kimberly, Dunworth William P, Gibbons Carrie L, Smithies Oliver, Caron Kathleen M
Department of Cell & Molecular Physiology, CB #7545, 6330 MBRB, 103 Mason Farm Rd., The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
Mol Cell Biol. 2006 Apr;26(7):2511-8. doi: 10.1128/MCB.26.7.2511-2518.2006.
Adrenomedullin (AM) is a multifunctional peptide vasodilator that is essential for life. To date, numerous in vitro studies have suggested that AM can mediate its biological effects through at least three different receptors. To determine the in vivo importance of the most likely candidate receptor, calcitonin receptor-like receptor, a gene-targeted knockout model of the gene was generated. Mice heterozygous for the targeted Calcrl allele appear normal, survive to adulthood, and reproduce. However, heterozygote matings fail to produce viable Calcrl-/- pups, demonstrating that Calcrl is essential for survival. Timed matings confirmed that Calcrl-/- embryos die between embryonic day 13.5 (E13.5) and E14.5 of gestation. The Calcrl-/- embryos exhibit extreme hydrops fetalis and cardiovascular defects, including thin vascular smooth muscle walls and small, disorganized hearts remarkably similar to the previously characterized AM-/- phenotype. In vivo assays of cellular proliferation and apoptosis in the hearts and vasculature of Calcrl-/- and AM-/- embryos support the concept that AM signaling is a crucial mediator of cardiovascular development. The Calcrl gene targeted mice provide the first in vivo genetic evidence that CLR functions as an AM receptor during embryonic development.
肾上腺髓质素(AM)是一种对生命至关重要的多功能肽类血管舒张剂。迄今为止,大量体外研究表明,AM可通过至少三种不同受体介导其生物学效应。为确定最有可能的候选受体——降钙素受体样受体在体内的重要性,构建了该基因的靶向敲除模型。靶向Calcr1等位基因的杂合小鼠外观正常,能存活至成年并繁殖。然而,杂合子交配未能产生存活的Calcr1-/-幼崽,这表明Calcr1对生存至关重要。定时交配证实,Calcr1-/-胚胎在妊娠第13.5天(E13.5)至E14.5天之间死亡。Calcr1-/-胚胎表现出严重的胎儿水肿和心血管缺陷,包括血管平滑肌壁薄以及心脏小且结构紊乱,与先前描述的AM-/-表型极为相似。对Calcr1-/-和AM-/-胚胎心脏和血管中的细胞增殖及凋亡进行的体内分析支持了这样的概念,即AM信号传导是心血管发育的关键介质。靶向Calcr1基因的小鼠提供了首个体内遗传学证据,表明CLR在胚胎发育过程中作为AM受体发挥作用。