Arjamaa O, Sormunen R, Lehto V P, Vuolteenaho O
Institute of Arctic Medicine, University of Oulu, Oulu, 90014, Finland.
Gen Comp Endocrinol. 2000 Dec;120(3):276-82. doi: 10.1006/gcen.2000.7558.
We have previously cloned and characterized a novel cardiac hormone from the salmon (Salmo salar) which has a uniquely heart-specific distribution and a low structural similarity with any other known natriuretic peptides. Specific antibodies were raised in goat against the salmon cardiac peptide. For localization and quantification, four different methods were applied: immunohistochemistry (avidin-biotin peroxidase), transmission electron microscopy, cryoimmunoelectron microscopy (protein A-gold), and a specific radioimmunoassay. Both atrial and ventricular myocytes stained immunohistochemically. The staining was similar in all myocytes and no specific myoendocrine cells were found. Within a single myocyte, both atrial and ventricular, the staining was stronger near the nucleus. Transmission electron microscopy revealed that both the atrium and the ventricle contained small sarcoplasmic granules of similar type with a diameter of 100 to 200 nm and an electron-dense core with a clear halo. The granules were typical vesicles which can be found in secretory cells utilizing the regulatory pathway. The highest number of granules was found near the nucleus, but granules were located also near the Golgi apparatus, between myofilament bundles, and in subsarcolemmal positions. Gold particles, conjugated to antibodies raised against the salmon cardiac peptide, were deposited on similar sarcoplasmic granules found in transmission electron microscopy. Among the sarcoplasmic granules with gold particles there were granules which did not show any cardiac peptide immunoreactivity. A significantly (Student's t test, P < 0.05) higher concentration of cardiac peptide was found in the heart atrium than in the ventricle, 16.2 +/- 3.5 pmol/mg tissue (n = 8) and 4.5 +/- 1.7 pmol/mg tissue (n = 8), respectively. The findings show that the salmon cardiac peptide is localized in secretory granules in both compartments of the heart. The morphology of the granules suggests that both the atrium and the ventricle utilize the regulatory pathway to release salmon cardiac peptide.
我们之前从鲑鱼(Salmo salar)中克隆并鉴定了一种新型心脏激素,它具有独特的心脏特异性分布,且与其他任何已知的利钠肽结构相似性较低。我们用山羊制备了针对鲑鱼心脏肽的特异性抗体。为了进行定位和定量,我们应用了四种不同的方法:免疫组织化学(抗生物素蛋白-生物素过氧化物酶法)、透射电子显微镜、冷冻免疫电子显微镜(蛋白A-金标法)以及特异性放射免疫测定法。心房和心室肌细胞在免疫组织化学染色中均呈阳性。所有肌细胞中的染色情况相似,未发现特异性心肌内分泌细胞。在单个心房和心室肌细胞内,细胞核附近的染色更强。透射电子显微镜显示,心房和心室均含有类似类型的小肌浆颗粒,直径为100至200纳米,有一个电子致密的核心和清晰的晕圈。这些颗粒是典型的囊泡,可在利用调节途径的分泌细胞中找到。颗粒数量最多的部位是细胞核附近,但在高尔基体附近、肌丝束之间以及肌膜下位置也有颗粒。与针对鲑鱼心脏肽产生的抗体结合的金颗粒沉积在透射电子显微镜下发现的类似肌浆颗粒上。在带有金颗粒的肌浆颗粒中,有些颗粒未显示任何心脏肽免疫反应性。通过学生t检验发现,心房中心脏肽的浓度显著(P < 0.05)高于心室,分别为16.2±3.5 pmol/mg组织(n = 8)和4.5±1.7 pmol/mg组织(n = 8)。研究结果表明,鲑鱼心脏肽定位于心脏两个腔室的分泌颗粒中。颗粒的形态表明,心房和心室都利用调节途径释放鲑鱼心脏肽。