Bauchet Anne-Laure, Masson Regis, Guffroy Magali, Slaoui Mohamed
Sanofi-Aventis R&D, DSAR, Preclinical Safety, Vitry-sur-Seine, France.
Toxicol Pathol. 2011 Dec;39(7):1115-28. doi: 10.1177/0192623311425060. Epub 2011 Oct 17.
Kidney is a major target organ in preclinical studies. In recent years, intense research has been undertaken to characterize novel renal toxicity biomarkers. In this context, we studied nephron segment specific antibodies against aquaporin-1 (AQP-1), α-glutathione-S-transferase (alpha-GST), Tamm-Horsfall protein (TH), calbindin-D(28K) (CalD), and aquaporin-2 (AQP-2), using an immunoperoxidase method on formalin-fixed paraffin-embedded kidney tissues of dogs, rats, mice, and Cynomolgus monkeys. AQP-1 was specific for proximal tubules and thin descending limbs of Henle's loops and AQP-2 for connecting and collecting ducts in dogs, rats, mice, and Cynomolgus monkeys. Alpha-GST stained the straight part of proximal tubules in dogs and proximal convoluted tubule and straight part of proximal tubules in rats. TH was specific for thick ascending limbs of Henle's loops in mice, rats, dogs, and Cynomolgus monkeys and stained additionally scattered cells in cortical connecting/collecting ducts of dogs. CalD was found in distal convoluted tubules and cortical connecting and collecting ducts of dogs, rats, and mice and in distal convoluted tubules, connecting ducts, and cortical and medullary collecting ducts of Cynomolgus monkey. This panel of antibodies may be a helpful tool to identify renal tubules by light microscopy in preclinical studies and to validate new biomarkers of renal toxicity.
肾脏是临床前研究中的主要靶器官。近年来,人们开展了大量研究以鉴定新型肾毒性生物标志物。在此背景下,我们采用免疫过氧化物酶法,对犬、大鼠、小鼠和食蟹猴经福尔马林固定、石蜡包埋的肾脏组织进行研究,检测针对水通道蛋白-1(AQP-1)、α-谷胱甘肽-S-转移酶(alpha-GST)、Tamm-Horsfall蛋白(TH)、钙结合蛋白-D(28K)(CalD)和水通道蛋白-2(AQP-2)的肾单位节段特异性抗体。在犬、大鼠、小鼠和食蟹猴中,AQP-1对近端小管和髓袢细降支具有特异性,AQP-2对连接管和集合管具有特异性。Alpha-GST在犬的近端小管直部以及大鼠的近端曲管和近端小管直部染色。TH在小鼠、大鼠、犬和食蟹猴的髓袢厚升支具有特异性,并且在犬的皮质连接/集合管中额外染色散在细胞。CalD在犬、大鼠和小鼠的远端曲管以及皮质连接管和集合管中发现,在食蟹猴的远端曲管、连接管以及皮质和髓质集合管中也有发现。这组抗体可能是临床前研究中通过光学显微镜识别肾小管以及验证新的肾毒性生物标志物的有用工具。