Asan E, Kugler P, Schiebler T H
Histochemistry. 1986;84(4-6):408-17. doi: 10.1007/BF00482971.
Sex-dependent protein handling in the rat renal tubular system was studied both qualitatively and quantitatively using the method of direct fluorescent protein tracing. The protein tracer, fluorescent ovalbumin, was synthesized by conjugating hen ovalbumin with fluorescein isothiocyanate (FITC), and the fluorescence characteristics of fluoresceinthiocarbamyl (FTC)-ovalbumin conjugates with different degrees of labelling were studied. Heavily labelled tracer was intravenously injected into male and female rats, and both kidneys were perfused; the right kidney was then homogenized and used for quantitative fluorometric measurements, while the left kidney was perfusion fixed and prepared for fluorescence microscopy. The tubular reabsorption of fluorescent ovalbumin was studied 4 min and 10 min after the injection of different doses (1.4, 7.0 and 14.0 mg/kg body weight) of the tracer, and the tubular catabolism was investigated in animals killed 60 and 120 min after the injection. Fluorescence microscopy demonstrated that, in both sexes and regardless of the dose administered and the time after injection, specifically fluorescent protein or its degradation products was only present in the epithelial cells of the proximal tubule. With regard to sex-dependent differences in protein handling, fluorometry indicated that at 4 min (7.0 mg) and at 10 min (all doses) after injection, female animals had reabsorbed more fluorescent protein than males. With regard to the catabolic phase, both the fluorescence microscopy and the fluorometric results showed that the female rats had degraded the fluorescent tracer at a significantly higher rate than males. The results are discussed in connection with sex-dependent proteinuria in rats.
采用直接荧光蛋白示踪法,对大鼠肾小管系统中性别依赖性蛋白处理进行了定性和定量研究。蛋白示踪剂荧光卵清蛋白是通过将鸡卵清蛋白与异硫氰酸荧光素(FITC)偶联合成的,并研究了不同标记程度的荧光硫代氨基甲酰(FTC)-卵清蛋白偶联物的荧光特性。将标记程度高的示踪剂静脉注射到雄性和雌性大鼠体内,对双侧肾脏进行灌注;然后将右肾匀浆用于荧光定量测量,而将左肾进行灌注固定并制备用于荧光显微镜检查。在注射不同剂量(1.4、7.0和14.0mg/kg体重)的示踪剂后4分钟和10分钟,研究荧光卵清蛋白的肾小管重吸收情况,并在注射后60分钟和120分钟处死动物,研究肾小管分解代谢情况。荧光显微镜检查表明,无论性别、给药剂量和注射后时间如何,特异性荧光蛋白或其降解产物仅存在于近端小管的上皮细胞中。关于蛋白处理的性别依赖性差异,荧光测定表明,在注射后4分钟(7.0mg)和10分钟(所有剂量)时,雌性动物重吸收的荧光蛋白比雄性动物多。关于分解代谢阶段,荧光显微镜检查和荧光测定结果均表明,雌性大鼠降解荧光示踪剂的速率明显高于雄性大鼠。结合大鼠性别依赖性蛋白尿对结果进行了讨论。