Lelongt B, Makino H, Kanwar Y S
Department of Pathology, Northwestern University Medical School, Chicago, Illinois.
Kidney Int. 1987 Oct;32(4):498-506. doi: 10.1038/ki.1987.238.
Maturation of extracellular matrices in relation to heparan sulfate proteoglycans was investigated during glomerular development. Antibodies directed against the core-protein (Mr = 18,000) of the basement membrane heparan sulfate-proteoglycan were utilized. The IgG fraction of the antibodies was conjugated with either fluorescein or rhodamine or 125I-iodine. The fluoresceinated and radioiodinated antibodies were given intravenously to three-day-old rats; their kidneys were obtained 6, 48, and 120 hours later and processed for immunohistochemical and tissue autoradiographic studies. At six hours (day 0), matrices of all the stages were labeled. The labeling was lowest in the vesicle stage and highest in the capillary stage. The autoradiographic grain-densities of vesicle, S-shaped, precapillary, and capillary stages were 10.62 +/- 0.54, 7.44 +/- 0.65, 11.37 +/- 1.03, and 18.35 +/- 1.97, respectively. After 48 hours (day 5), the grain-density for S-shaped body decreased from 7.44 +/- 0.65 to 4.38 +/- 0.61, while no appreciable change in the precapillary and capillary stages was observed. At this time interval, the fluorescein-labeled glomeruli were seen in the deeper cortex only. After 120 hours (day 8), the grain-density for the capillary stage decreased from 18.35 +/- 1.97 to 2.82 +/- 0.92. The double-labeling experiments included a second injection of rhodaminated antibody at 48 hours (day 5). In these experiments, newly developed glomeruli of varying stages were seen in the superficial cortex. With the use of double filters, the immature glomeruli in the superficial cortex and mature ones in the deeper cortex were visualized.(ABSTRACT TRUNCATED AT 250 WORDS)
在肾小球发育过程中,研究了与硫酸乙酰肝素蛋白聚糖相关的细胞外基质的成熟情况。使用了针对基底膜硫酸乙酰肝素蛋白聚糖核心蛋白(分子量 = 18,000)的抗体。抗体的IgG部分与荧光素、罗丹明或125I碘结合。将荧光素标记和放射性碘标记的抗体静脉注射给3日龄大鼠;6、48和120小时后获取它们的肾脏,并进行免疫组织化学和组织放射自显影研究。6小时(第0天)时,所有阶段的基质都被标记。在囊泡阶段标记最低,在毛细血管阶段最高。囊泡、S形、毛细血管前和毛细血管阶段的放射自显影颗粒密度分别为10.62±0.54、7.44±0.65、11.37±1.03和18.35±1.97。48小时(第5天)后,S形体的颗粒密度从7.44±0.65降至4.38±0.61,而毛细血管前和毛细血管阶段未观察到明显变化。在此时间间隔,仅在较深皮质中可见荧光素标记的肾小球。120小时(第8天)后,毛细血管阶段的颗粒密度从18.35±1.97降至2.82±0.92。双重标记实验包括在48小时(第5天)第二次注射罗丹明标记的抗体。在这些实验中,在浅表皮质中可见不同阶段新发育的肾小球。使用双重滤光片,可以观察到浅表皮质中的未成熟肾小球和较深皮质中的成熟肾小球。(摘要截断于250字)