Cantin M, Gutkowska J, Thibault G, Milne R W, Ledoux S, MinLi S, Chapeau C, Garcia R, Hamet P, Genest J
Histochemistry. 1984;80(2):113-27. doi: 10.1007/BF00679984.
Antibodies produced in the mouse by repeated intraperitoneal injections of partly purified atrial natriuretic factor (low molecular weight peptide (LMWP) and high molecular weight peptide (HMWP)) have been used to localize these factors by immunohistochemistry (immunofluorescence and immunoperoxidase method) and by immunocytochemistry (protein A-gold technique) in the heart of rats and of a variety of animal species including man and in the rat salivary glands. Immunofluorescence and the immunoperoxidase method gave identical results; in the rat, atrial cardiocytes gave a positive reaction at both nuclear poles while ventricular cardiocytes were consistently negative. The cardiocytes of the right atrial appendage were more intensely reactive than those localized in the left appendage. A decreasing gradient of intensity was observed from the subpericardial to the subendocardial cardiocytes. The cardiocytes of the interatrial septum were only lightly granulated. Sodium deficiency and thirst (deprivation of drinking water for 5 days) produced, as already shown at the ultrastructural level, a marked increase in the reactivity of all cardiocytes from both atria with the same gradient of intensity as in control animals. Cross-reactivity of intragranular peptides with the rat antibodies allowed visualization of specific granules in a variety of animal species (mouse, guinea pig, rabbit, rat, dog) and in human atrial appendages. No reaction could be elicited in the frog atrium and ventricle although, in this species, specific granules have been shown to be present by electron microscopy in all cardiac chambers. With the protein A-gold technique, at the ultrastructural level, single labeling (use of one antibody on one face of a fine section) or double labeling (use of two antibodies on the two faces of a fine section) showed that the two peptides are localized simultaneously in all three types (A, B and D) of specific granules. In the rat salivary glands, immunofluorescence and the immunoperoxidase method showed reactivity exclusively in the acinar cells. The reaction was most intense in the acinar cells of the parotid gland. In the sublingual gland, only the serous cells, sometimes forming abortive "demi-lunes", were reactive. In the submaxillary gland, the reaction was weaker and distributed seemingly haphazardly in the gland. The most constantly reactive cells were localized near the capsule while many cells did not contain visible reaction product.
通过反复腹腔注射部分纯化的心房利钠因子（低分子量肽（LMWP）和高分子量肽（HMWP））在小鼠体内产生的抗体，已被用于通过免疫组织化学（免疫荧光和免疫过氧化物酶法）以及免疫细胞化学（蛋白A-金技术）在大鼠以及包括人类在内的多种动物物种的心脏和大鼠唾液腺中定位这些因子。免疫荧光和免疫过氧化物酶法得出了相同的结果；在大鼠中，心房心肌细胞在两个核极均呈阳性反应，而心室心肌细胞始终为阴性。右心耳的心肌细胞比左心耳中的心肌细胞反应更强。从心包下心肌细胞到心内膜下心肌细胞观察到强度递减梯度。房间隔的心肌细胞颗粒化程度较轻。如在超微结构水平上已经显示的那样，缺钠和口渴（剥夺饮用水5天）使两个心房所有心肌细胞的反应性显著增加，其强度梯度与对照动物相同。颗粒内肽与大鼠抗体的交叉反应性使得能够在多种动物物种（小鼠、豚鼠、兔子、大鼠、狗）以及人类心耳中观察到特定颗粒。尽管在该物种中通过电子显微镜已显示所有心腔中均存在特定颗粒，但在蛙的心房和心室中未引发反应。使用蛋白A-金技术，在超微结构水平上，单标记（在薄切片的一个面上使用一种抗体）或双标记（在薄切片的两个面上使用两种抗体）表明这两种肽同时定位于所有三种类型（A、B和D）的特定颗粒中。在大鼠唾液腺中，免疫荧光和免疫过氧化物酶法显示仅腺泡细胞有反应性。反应在腮腺的腺泡细胞中最为强烈。在舌下腺中，只有有时形成不完全“半月形”的浆液性细胞有反应性。在颌下腺中，反应较弱，且在腺体内似乎呈随机分布。反应最恒定的细胞位于被膜附近，而许多细胞不含可见的反应产物。
