A quantitative analysis of the endocytic pathway in baby hamster kidney cells.
A quantitative analysis of the endocytic pathway in baby hamster kidney cells.
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DOI：
10.1083/jcb.109.6.2703
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发表时间：
1989-12
期刊：
影响因子：
--
通讯作者：
Marsh M
中科院分区：
文献类型：
--
作者：
Griffiths G;Back R;Marsh M
用辣根过氧化物酶（HRP）作液相标记，对幼仓鼠肾（BHK）细胞内吞途径的各个部分进行了形态学分析。内吞作用次数增加后标记的内吞结构已被确定，并测量其体积和表面密度。在第一个2分钟的HRP摄取的标记结构的体积密度迅速增加，此后保持不变，为未来13-18分钟。这个平台表示内体细胞器的体积密度和占0.65%的细胞质体积（或6.8微米3每个细胞）。标记的结构由管状池元件组成，其经常与300-400 nm的囊泡连续观察。内化15-20分钟后，HRP标记结构的体积密度再次迅速增加，并在标记的30和60分钟之间达到第二个平台。这第二次增加对应于可检测水平的HRP到达后，酸性磷酸酶（Acetylase）反应的车厢。这些结构，包括前溶酶体和溶酶体，主要是囊泡和共同占3.5%的细胞质体积（或37微米3每个细胞）。两类细胞器（内体和前溶酶体/溶酶体）的绝对外周表面积估计分别为430和370 μ m 2/细胞。在摄取的前2分钟内内化的液体体积比此时标记的隔室体积小5 - 7倍。为了解释这些结果，我们提出，从细胞表面摄取后，HRP被传递到，并稀释，内体是预先存在的细胞器，最初没有标记。随着内吞作用时间的增加，早期内体中HRP的浓度增加，因为更多的标记物进入该隔室。在冷冻切片上使用抗HRP抗体和胶体金直接显示在早期时间点期间内体中HRP浓度的升高。在本研究中给出的体视学值，结合早期的研究，提供了一个最小的估计膜的总表面积和膜的合成率在BHK细胞。
A morphological analysis of the compartments of the endocytic pathway in baby hamster kidney (BHK) cells has been made using the fluid-phase marker horseradish peroxidase (HRP). The endocytic structures labeled after increasing times of endocytosis have been identified and their volume and surface densities measured. In the first 2 min of HRP uptake the volume density of the labeled structures increased rapidly and thereafter remained constant for the next 13-18 min. This plateau represents the volume density of endosome organelles and accounts for 0.65% of the cytoplasmic volume (or 6.8 microns 3 per cell). The labeled structures consist of tubular-cisternal elements which are frequently observed in continuity with 300-400 nm vesicles. After 15-20 min of internalization the volume density of HRP-labeled structures again increased rapidly and reached a second plateau between 30 and 60 min of labeling. This second increase corresponded to detectable levels of HRP reaching later, acid phosphatase (AcPase)-reactive compartments. These structures, comprising the prelysosomes and lysosomes, were mostly vesicular and collectively accounted for 3.5% of the cytoplasmic volume (or 37 microns 3 per cell). The absolute peripheral surface areas of the two classes of organelles (endosomes and prelysosomes/lysosomes) were estimated to be 430 and 370 microns 2 per cell, respectively. The volume of fluid internalized in the first 2 min of uptake was five- to sevenfold less than the volume of the compartment labeled in this time. To account for these results we propose that, after uptake from the cell surface, HRP is delivered to, and diluted in, endosomes that are preexisting organelles initially devoid of the marker. With increasing times of endocytosis the concentration of HRP in the early endosomes increases, as more of the marker enters this compartment. An elevation in HRP concentration in endosomes during the early time points was shown directly using anti- HRP antibodies and colloidal gold on cryosections. The stereological values given in the present study, in combination with earlier studies, provide a minimum estimate for both the total surface area of membranes and the rate of membrane synthesis in a BHK cell.
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DOI：
10.1007/bf00307161
发表时间：
1973-01-01
期刊：
ZEITSCHRIFT FUR ZELLFORSCHUNG UND MIKROSKOPISCHE ANATOMIE
影响因子：
--
作者：
COPE, GH;WILLIAMS, MA
通讯作者：
WILLIAMS, MA
影响因子：
11.4
作者：
KIELIAN, MC;MARSH, M;HELENIUS, A
通讯作者：
HELENIUS, A
影响因子：
5.6
作者：
MARSH, M;HELENIUS, A
通讯作者：
HELENIUS, A
影响因子：
7.8
作者：
GRIFFITHS, G;QUINN, P;WARREN, G
通讯作者：
WARREN, G
影响因子：
5.6
作者：
GREEN, J;GRIFFITHS, G;WARREN, G
通讯作者：
WARREN, G