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嵌合小鼠中β-葡萄糖醛酸酶的细胞间转移

Intercellular transfer of beta-glucuronidase in chimeric mice.

作者信息

Herrup K, Mullen R J

出版信息

J Cell Sci. 1979 Dec;40:21-31. doi: 10.1242/jcs.40.1.21.

Abstract

Chimeric mice were produced by aggregating 2 embryos, each of which was homozygous for a different structural allele of the enzyme beta-glucuronidase. The alleles used were Gusb, the 'wild-type' allele, and Gush, an allele whose gene product shows decreased activity in all tissues as well as decreased heat stability. Staining of untreated adult chimeric livers for glucuronidase activity revealed a mosaic of high (Gusb) and low (Gush) activity cells. The boundaries between cells of different activity were sharp and revealed no diffusion of enzyme or reaction product. Treating sections at 73 degrees c before staining led to a decay of staining activity in normal (non-chimeric) Gush/Gush and Gusb/Gusb tissue. The rates of activity loss under the conditions used differed by 10-fold. Of the 2 genotypes in the chimera, the dark-staining, Gusb, cells decayed in a fashion similar to that of the Gusb/Gusb control. The light-staining, Gush, cells of the chimera lost their staining a a unique fashion. Within 20 min they quickly lost a majority of their staining activity but that which remained was relatively heat stable. The second Gusb-like phase of the decay, seen both photographically and photometrically, suggests that Gusb gene product has been transferred to cells of Gush/Gush genotype.

摘要

通过聚合两个胚胎培育出嵌合小鼠,每个胚胎对于β-葡萄糖醛酸酶的不同结构等位基因都是纯合的。所使用的等位基因是“野生型”等位基因Gusb和Gush,后者的基因产物在所有组织中的活性均降低,且热稳定性也降低。对未经处理的成年嵌合肝脏进行葡萄糖醛酸酶活性染色,结果显示出高活性(Gusb)和低活性(Gush)细胞的镶嵌图案。不同活性细胞之间的边界清晰,未发现酶或反应产物的扩散现象。在染色前于73摄氏度处理切片,导致正常(非嵌合)Gush/Gush和Gusb/Gusb组织中的染色活性衰减。在所使用的条件下,活性丧失的速率相差10倍。在嵌合体的两种基因型中,深色染色的Gusb细胞的衰减方式与Gusb/Gusb对照相似。嵌合体中浅色染色的Gush细胞以独特的方式失去其染色。在20分钟内,它们迅速丧失了大部分染色活性,但剩余的活性相对热稳定。从照片和光度测量中都能看到的衰减的第二个类似Gusb的阶段,表明Gusb基因产物已转移到Gush/Gush基因型的细胞中。

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