Dousa T P, Barnes L D, Ong S H, Steiner A L
Proc Natl Acad Sci U S A. 1977 Aug;74(8):3569-73. doi: 10.1073/pnas.74.8.3569.
Adenosine 3':5'-cyclic monophosphate (cAMP) and guanosine 3':5'-cyclic monophosphate (cGMP) were localized in cells of rat kidney cortex by an immunocytochemical technique before and after perfusion with parathyroid hormone (PTH). In control tissues the cAMP antiserum detected approximately the same intensity of fluorescence in cytoplasmic epithelial cell elements of cortical tubules and glomeruli (cells of Bowman's capsule and podocytes). PTH increased fluorescence in these glomerular cells and increased cAMP fluorescence in cytoplasmic granules in proximal tubular cells. These granules, whose structure has not been identified, were located predominantly on the luminal side of the tubular cells. In control rats, the renal cortical fluorescence detected with the cGMP antiserum was more pronounced in glomeruli (predominantly in the mesangial areas) and lesser amounts of fluorescence were observed in tubules. After PTH treatment, cGMP fluorescence increased in glomeruli and in renal tubular cells. A bright linear pattern of fluorescence was found in the area of the tubular luminal membrane. Perfusion with PTH caused relatively small increases in total tissue cAMP and no consistent increases in total tissue cGMP. Our observations suggest that both cAMP and cGMP are involved in the glomerular and tubular responses to PTH and point out the added dimension that this immunocytochemical technique brings to studies of cyclic nucleotide dynamics in heterogeneous tissues.
采用免疫细胞化学技术,在灌注甲状旁腺激素(PTH)前后,对大鼠肾皮质细胞中的3':5'-环磷酸腺苷(cAMP)和3':5'-环磷酸鸟苷(cGMP)进行定位。在对照组织中,cAMP抗血清在皮质肾小管和肾小球的细胞质上皮细胞成分(鲍曼囊细胞和足细胞)中检测到大致相同强度的荧光。PTH增加了这些肾小球细胞中的荧光,并增加了近端肾小管细胞细胞质颗粒中的cAMP荧光。这些结构尚未明确的颗粒主要位于肾小管细胞的管腔侧。在对照大鼠中,用cGMP抗血清检测到的肾皮质荧光在肾小球中更明显(主要在系膜区),而在肾小管中观察到的荧光较少。PTH处理后,肾小球和肾小管细胞中的cGMP荧光增加。在肾小管腔膜区域发现了明亮的线性荧光模式。灌注PTH导致组织总cAMP相对小幅增加,而组织总cGMP没有持续增加。我们的观察结果表明,cAMP和cGMP都参与了肾小球和肾小管对PTH的反应,并指出了这种免疫细胞化学技术为异质组织中环状核苷酸动力学研究带来的新维度。