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高尔基体中的铅染色。

Lead staining in the Golgi complex.

作者信息

McClintock J, Locke M

出版信息

Tissue Cell. 1982;14(3):541-54. doi: 10.1016/0040-8166(82)90046-5.

Abstract

Lead ions at similar concentrations to those used for Gomori type phosphatase localization stain some parts of the vacuolar system, particularly compartments of the Golgi complex (GC) and isolation envelopes (im) in a characteristic way in both vertebrates and invertebrates. After fixation in 2.5% glutaraldehyde, lead citrate in acetate or aspartate buffer (pH 5.5-7.2) leaves the contents of GC cisternal compartments with a fine particulate stippling. In the fat body of Calpodes ethlius and in mouse pancreas the staining is faint but definite without further enhancement of contrast, although it is easily overlooked after section staining. The distribution of lead stain differs from that of the lead phosphate precipitated after Gomori type acid phosphatase reactions. Whereas lead stain may be in all GC and im compartments, acid phosphatase is restricted to the innermost saccules and nearby vacuoles. The compartment specific staining by led also differs from the generalized staining in all compartments given by uranyl. Thus the contents of luminal membrane surfaces of some parts of the vacuolar system can be characterized by their ability to bind lead. In cells where protein synthesis has been blocked by cycloheximide, secretory vesicles are absent and the RER and GC from the generalized staining in all compartments given by uranyl. Thus the contents of luminal membrane surfaces of some parts of the vacuolar system can be characterized by their ability to bind lead. In cells where protein synthesis has been blocked by cycloheximide, secretory vesicles are absent and the RER and GC from the generalized staining in all compartments given by uranyl. Thus the contents of luminal membrane surfaces of some parts of the vacuolar system can be characterized by their ability to bind lead. In cells where protein synthesis has been blocked by cycloheximide, secretory vesicles are absent and the RER and GC cisternae are devoid of uranyl stainable material. However, lead staining and acid phosphatase activity in the GC continue. We presume that they mark the environment within these cisternae rather than the proteins passing through them. This environment is itself not static. Several observations suggest that the function of cisternae that is detectable by lead staining is temporally discontinuous and related to a stage of maturation or development. Only early stage ims stain: the staining ceases by the beginning of autophagy after hydrolytic enzymes are presumed to have been added. Condensing vacuoles cease to stain as the central core crystallizes out. Stain may be absent from one or two GC saccules at any position in the stack as though the phase of lead staining (or lack or it) can move progressively through the system. We conclude that in studies characterizing components of the vacuolar system it is necessary to separate those that mark transient occupants of a compartment from those that mark the compartment itself. Both may vary temporally independently from one another.

摘要

与用于Gomori型磷酸酶定位的铅离子浓度相似时,铅会以一种特征性方式对脊椎动物和无脊椎动物的液泡系统的某些部分进行染色,特别是高尔基体复合体(GC)的隔室和隔离膜(im)。在2.5%戊二醛中固定后,醋酸盐或天冬氨酸盐缓冲液(pH 5.5 - 7.2)中的柠檬酸铅会使GC潴泡隔室的内容物呈现出细微的颗粒状斑点。在埃氏角斑蛾的脂肪体和小鼠胰腺中,染色较淡但很明显,无需进一步增强对比度,不过在切片染色后很容易被忽略。铅染色的分布与Gomori型酸性磷酸酶反应后沉淀的磷酸铅的分布不同。虽然铅染色可能存在于所有GC和im隔室中,但酸性磷酸酶仅限于最内层的囊泡和附近的液泡。铅的隔室特异性染色也不同于铀酰对所有隔室的普遍染色。因此,液泡系统某些部分的腔膜表面内容物可以通过它们结合铅的能力来表征。在蛋白质合成被环己酰亚胺阻断的细胞中,分泌小泡不存在,粗面内质网和GC潴泡也没有铀酰可染色的物质。然而,GC中的铅染色和酸性磷酸酶活性仍在继续。我们推测它们标记的是这些潴泡内的环境,而不是穿过它们的蛋白质。这种环境本身并非静止不变。一些观察结果表明,可通过铅染色检测到的潴泡功能在时间上是不连续的,并且与成熟或发育阶段相关。只有早期的im会染色:在假定水解酶已添加后,自噬开始时染色停止。随着中央核心结晶析出,浓缩液泡停止染色。在堆叠中任何位置的一个或两个GC囊泡可能没有染色,就好像铅染色的阶段(或缺乏染色)可以在系统中逐渐移动。我们得出结论,在表征液泡系统成分的研究中,有必要将标记隔室短暂占据者的成分与标记隔室本身的成分区分开来。两者在时间上可能彼此独立变化。

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