Shields P P, Glembotski C C
Biochem Biophys Res Commun. 1987 Jul 31;146(2):547-53. doi: 10.1016/0006-291x(87)90563-8.
Although cultures of neonatal rat atria and ventricles have been widely used to study ANP biosynthesis and secretion, little is known regarding the circulating form of ANP in neonatal animals. To begin to address this issue, we have developed a method for perfusing isolated neonatal rat hearts. Reversed phase-HPLC analysis of the heart effluents coupled with ANP RIA demonstrated that the predominant form of ANP released was chromatographically identical to ANP(99-126). Size exclusion-HPLC confirmed that the secreted ANP was indistinguishable from ANP(99-126). This demonstrated that the neonatal rat heart can efficiently generate and secrete a peptide similar to the circulating form of ANP found in adult rats, and further justifies the use of neonatal rat atria as a source of primary cells for studies of ANP biosynthesis and secretion.
尽管新生大鼠心房和心室的培养物已被广泛用于研究心钠素(ANP)的生物合成和分泌,但对于新生动物体内循环形式的心钠素却知之甚少。为了开始解决这个问题,我们开发了一种灌注新生大鼠离体心脏的方法。对心脏流出液进行反相高效液相色谱(HPLC)分析并结合心钠素放射免疫分析(RIA)表明,释放的心钠素的主要形式在色谱上与心钠素(99 - 126)相同。尺寸排阻高效液相色谱证实,分泌的心钠素与心钠素(99 - 126)无法区分。这表明新生大鼠心脏能够有效地生成和分泌一种与成年大鼠体内循环形式的心钠素相似的肽,并且进一步证明了将新生大鼠心房作为研究心钠素生物合成和分泌的原代细胞来源的合理性。