Shirataki S, Takihara H, Imoto K, Kamada K, Baba Y, Naito K
Department of Urology, Yamaguchi University School of Medicine, Ube, Japan.
Andrologia. 1994 Nov-Dec;26(6):343-9. doi: 10.1111/j.1439-0272.1994.tb00814.x.
The utility of the 5-bromodeoxyuridine (BrdUrd) labelling technique for the quantitative analysis of spermatogenic deoxyribonucleic acid (DNA) synthesis was investigated in the rat. Rat testicles were labelled by a single intraperitoneal injection of 100 mg kg-1 of BrdUrd. The testicles were removed 1 h after injection, fixed in Bouin's fluid and embedded in paraffin. BrdUrd-labelled cells were detected by immunohistochemical staining using a monoclonal anti-BrdUrd antibody. The number of BrdUrd-labelled tubules per total number of tubules (percent L.T.), the number of BrdUrd-labelled cells per total number of tubules (tubular ratio) and the number of BrdUrd-labelled cells per number of Sertoli cells (Sertoli cell ratio in BrdUrd-labelled cells) were calculated as indices of spermatogenic DNA synthesis during each stage of the seminiferous epithelial wave. BrdUrd labelling was found exclusively in the nuclei of spermatogonia and in preleptotene spermatocytes in the seminiferous epithelium. The percent L.T. was generally greater than 50%, except in stages VI, VII and XIV, and the tubular as well as Sertoli cell ratios in BrdUrd-labelled cells was greater than 2.0 and 0.15, respectively, in stages I, II-III, V, VIII, X, and XII. The tubular ratio and Sertoli cell ratio in BrdUrd-labelled cells along the seminiferous epithelial wave had two distinct peaks. The distribution of the tubular ratio using the BrdUrd-labelling technique correlated well with the distribution previously established by measuring tritiated thymidine uptake per tubule. Thus, the BrdUrd labelling technique, which is more efficient than the tritiated thymidine labelling technique, can be used to quantitatively evaluate spermatogenic DNA synthesis.
在大鼠中研究了5-溴脱氧尿苷（BrdUrd）标记技术用于定量分析生精脱氧核糖核酸（DNA）合成的效用。通过腹腔内单次注射100mg/kg的BrdUrd对大鼠睾丸进行标记。注射后1小时取出睾丸，固定于Bouin氏液中并包埋于石蜡中。使用单克隆抗BrdUrd抗体通过免疫组织化学染色检测BrdUrd标记的细胞。计算每曲细精管总数中BrdUrd标记的曲细精管数量（标记曲细精管百分比，L.T.%）、每曲细精管总数中BrdUrd标记的细胞数量（曲细精管比率）以及每支持细胞数量中BrdUrd标记的细胞数量（BrdUrd标记细胞中的支持细胞比率），作为生精上皮波各阶段生精DNA合成的指标。发现BrdUrd标记仅存在于生精上皮中生精细胞的细胞核和前细线期精母细胞中。除了VI、VII和XIV期外，L.T.%一般大于50%，在I、II-III、V、VIII、X和XII期，BrdUrd标记细胞中的曲细精管比率和支持细胞比率分别大于2.0和0.15。沿生精上皮波的BrdUrd标记细胞中的曲细精管比率和支持细胞比率有两个明显的峰值。使用BrdUrd标记技术的曲细精管比率分布与先前通过测量每曲细精管的氚标记胸腺嘧啶摄取所确定的分布密切相关。因此，比氚标记胸腺嘧啶标记技术更有效的BrdUrd标记技术可用于定量评估生精DNA合成。
