Herzog V, Farquhar M G
Proc Natl Acad Sci U S A. 1977 Nov;74(11):5073-7. doi: 10.1073/pnas.74.11.5073.
Dextran was used to trace membrane retrieved from the luminal surface after induced exocytosis in secretory cells of rat lacrimal and parotid glands. Two different approaches were used: (a) isolated acini were incubated in vitro with dextran followed by stimulation with carbamylcholine (lacrimal) or isoproterenol (parotid) and (b) rats were injected with isoproterenol followed by dextran infusion into the parotid duct in vivo. The main findings were the same regardless of the gland source or experimental approach. Dextran was taken up initially via coated pits into smooth-surfaced apical vesicles. Shortly thereafter it was found in multiple cell compartments: within the stacked Golgi cisternae, in condensing vacuoles, and in lysosomes. Uptake was more rapid and uniform in vivo; dextran was seen in multiple cisternae of numerous Golgi complexes within 5 min after infusion. In acini incubated in vitro uptake into Golgi cisternae was more delayed and occurred with increasing frequency up to 60 min; also, more dextran was taken up into lysosomes, which were more numerous in vitro than in vivo. The results demonstrate that, after exocytosis, membrane is removed from the cell surface via vesicles that fuse with multiple cell compartments. The two novel findings are: (a) the demonstration that the tracer can reach most of the Golgi cisternae in a given stack and (b) the demonstration of the rapidity with which the process takes place (i.e., within 5 min). The findings imply that at least some membrane retrieved from the cell surface after exocytosis fuses with the stacked Golgi cisternae.
葡聚糖被用于追踪大鼠泪腺和腮腺分泌细胞诱导胞吐后从管腔表面回收的膜。采用了两种不同的方法:(a) 将分离的腺泡与葡聚糖在体外孵育,然后用氨甲酰胆碱(泪腺)或异丙肾上腺素(腮腺)刺激;(b) 给大鼠注射异丙肾上腺素,然后在体内将葡聚糖注入腮腺导管。无论腺体来源或实验方法如何,主要发现都是相同的。葡聚糖最初通过被膜小窝进入表面光滑的顶端小泡。此后不久,在多个细胞区室中发现了它:在堆叠的高尔基池、浓缩泡和溶酶体中。体内摄取更快且更均匀;输注后5分钟内,在众多高尔基复合体的多个池内可见葡聚糖。在体外孵育的腺泡中,进入高尔基池的摄取更延迟,并且频率逐渐增加,直至60分钟;此外,更多的葡聚糖被摄取到溶酶体中,溶酶体在体外比在体内更多。结果表明,胞吐后,膜通过与多个细胞区室融合的小泡从细胞表面被移除。两个新发现是:(a) 证明示踪剂可以到达给定堆叠中的大多数高尔基池;(b) 证明该过程发生的速度(即5分钟内)。这些发现意味着,胞吐后从细胞表面回收的至少一些膜与堆叠的高尔基池融合。