Simionescu N, Simionescu M, Palade G E
J Cell Biol. 1972 May;53(2):365-92. doi: 10.1083/jcb.53.2.365.
The pathway followed by macromolecules across the wall of visceral capillaries has been studied by using a set of tracers of graded sizes, ranging in diameter from 100 A (ferritin) to 300 A (glycogen). Polysaccharide particles, i.e. dextran 75 (mol wt approximately 75,000; diam approximately 125 A), dextran 250 (mol wt 250,000; diam approximately 225 A), shellfish glycogen (diam approximately 200 A) and rabbit liver glycogen (diam approximately 300 A), are well tolerated by Wistar-Furth rats and give no vascular reactions ascribable to histamine release. Good definition and high contrast of the tracer particles were obtained in a one-step fixation-in block staining of the tissues by a mixture containing aldehydes, OsO(4) and lead citrate in phosphate or arsenate buffer, pH 7.4, followed by lead staining of sections. The glycogens and dextrans used move out of the plasma through the fenestrae and channels of the endothelium relatively fast (3-7 min) and create in the pericapillary spaces transient (2-5 min) concentration gradients centered on the fenestrated sectors of the capillary walls. The tracers also gained access to the plasmalemmal vesicles, first on the blood front and subsequently on the tissue front of the endothelium. The particles are temporarily retained by the basement membrane. No probe moved through the intercellular junctions. It is concluded that, in visceral capillaries, the fenestrae, channels, and plasmalemmal vesicles, viewed as related parts in a system of dynamic structures, are the structural equivalent of the large pore system.
通过使用一组大小分级的示踪剂,研究了大分子穿过内脏毛细血管壁的途径,这些示踪剂的直径范围从100埃(铁蛋白)到300埃(糖原)。多糖颗粒,即右旋糖酐75(分子量约75,000;直径约125埃)、右旋糖酐250(分子量250,000;直径约225埃)、贝类糖原(直径约200埃)和兔肝糖原(直径约300埃),Wistar-Furth大鼠对其耐受性良好,且不会因组胺释放而产生血管反应。通过在pH 7.4的磷酸盐或砷酸盐缓冲液中含有醛、OsO(4)和柠檬酸铅的混合物对组织进行一步固定包埋染色,然后对切片进行铅染色,获得了示踪剂颗粒的良好清晰度和高对比度。所使用的糖原和右旋糖酐通过内皮细胞的窗孔和通道相对较快地(3 - 7分钟)从血浆中移出,并在毛细血管周围间隙中以毛细血管壁有窗孔部分为中心形成短暂的(2 - 5分钟)浓度梯度。示踪剂也进入了质膜小泡,首先是在内皮细胞的血液侧,随后是在组织侧。颗粒被基底膜暂时保留。没有探针穿过细胞间连接。得出的结论是,在内脏毛细血管中,窗孔、通道和质膜小泡,被视为动态结构系统中的相关部分,是大孔系统的结构等效物。