Fadool J M, Aggarwal S K
Department of Zoology, Michigan State University, East Lansing 48824.
J Histochem Cytochem. 1990 Jan;38(1):7-12. doi: 10.1177/38.1.2403579.
We investigated the immunoperoxidase demonstration of vasopressin (VSP) bound to paraffin-embedded sections of rat kidney and the effects of various fixatives. Slices of rat kidney from normal and 4-day water-deprived rats were incubated with 10(-7) M VSP, fixed, and embedded in paraffin. Hydrated sections of these tissues were again incubated with 10(-7) M VSP or 10(-7) M VSP and 10(-5) M oxytocin (OXY). VSP bound to the sections was demonstrated using rabbit anti-Arg8 VSP antiserum and peroxidase-labeled second antibody. In sections of kidney from both normal and water-deprived rats, immunoperoxidase labeling was most intense in the renal papilla and was restricted to the cells of the ducts of Bellini and loops of Henle. In the medulla, the collecting ducts and medullary thick ascending limbs of Henle were moderately stained. In the normal kidney sections there was no staining of the proximal tubules, distal convoluted tubules (DCT), and only slight staining of the cortical collecting ducts (CCD). However, in the water-deprived rats there was a considerable increase in the staining of the DCT and CCD. Simultaneous incubation in OXY and VSP resulted in reduced immunoperoxidase labeling of the tubules. Omission of VSP incubation led to a similar decrease in stain intensity, indicating a specificity for the sites of VSP binding. This technique allows the identification of cells responsible for the binding of VSP in the kidney.
我们研究了抗利尿激素（VSP）与大鼠肾脏石蜡包埋切片结合的免疫过氧化物酶显示法以及各种固定剂的作用。将正常大鼠和禁水4天大鼠的肾脏切片与10⁻⁷ M VSP孵育，固定后石蜡包埋。这些组织的水化切片再次与10⁻⁷ M VSP或10⁻⁷ M VSP和10⁻⁵ M催产素（OXY）孵育。使用兔抗精氨酸⁸ VSP抗血清和过氧化物酶标记的二抗显示结合到切片上的VSP。在正常大鼠和禁水大鼠的肾脏切片中，免疫过氧化物酶标记在肾乳头处最为强烈，且仅限于贝利尼管和亨利袢的细胞。在髓质中，集合管和髓质部亨利袢升支被中度染色。在正常肾脏切片中，近端小管、远端曲管（DCT）无染色，皮质集合管（CCD）仅有轻微染色。然而，在禁水大鼠中，DCT和CCD的染色有相当程度的增加。同时用OXY和VSP孵育导致小管的免疫过氧化物酶标记减少。省略VSP孵育导致染色强度有类似降低，表明对VSP结合位点具有特异性。该技术可鉴定肾脏中负责VSP结合的细胞。
