Sun X Z, Harada Y N, Takahashi S, Shiomi N, Shiomi T
The 4th Research Group, National Institute of Radiological Sciences, Chiba, Japan.
J Neurosci Res. 2001 May 15;64(4):348-54. doi: 10.1002/jnr.1085.
Laboratory mice carrying the nonfunctional xeroderma pigmentosum group G gene (the mouse counterpart of the human XPG gene) alleles have been generated by using gene-targeting and embryonic stem cell technology. Homozygote animals of this autosomal recessive disease exhibited signs and symptoms, such as postnatal growth retardation, reduced levels of activity, progressive ataxia and premature death, similar to the clinical manifestations of Cockayne syndrome (CS). Histological analysis of the cerebellum revealed multiple pyknotic cells in the Purkinje cell layer of the xpg homozygotes, which had atrophic cell bodies and shrunken nuclei. Further examination by an immunohistochemistry for calbindin-D 28k (CaBP) showed that a large number of immunoreactive Purkinje cells were atrophic and their dendritic trees were smaller and shorter than in wild-type littermates. These results indicated a marked degeneration of Purkinje cells in the xpg mutant cerebellum. Study by in situ detection of DNA fragmentation in the cerebellar cortex demonstrated that some deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin in situ nick labeling (TUNEL)-positive cells appeared in the granule layer of the mutant mice, but few cell deaths were confirmed in the Purkinje layer. These results suggested Purkinje cell degeneration in the mutant cerebellum was underway, in which much Purkinje cell death had not appeared, and the appearance of some abnormal cerebellar symptoms in the xpg-deficient mice was not only due to a marked Purkinje cell degeneration, but also to damage of other cells.
通过基因靶向和胚胎干细胞技术,已培育出携带无功能的着色性干皮病G组基因(人类XPG基因的小鼠对应基因)等位基因的实验小鼠。这种常染色体隐性疾病的纯合子动物表现出一些体征和症状,如出生后生长迟缓、活动水平降低、进行性共济失调和过早死亡,类似于科凯恩综合征(CS)的临床表现。对小脑的组织学分析显示,xpg纯合子的浦肯野细胞层中有多个固缩细胞,其细胞体萎缩,细胞核缩小。通过对钙结合蛋白-D 28k(CaBP)进行免疫组织化学进一步检查发现,大量免疫反应阳性的浦肯野细胞萎缩,其树突比野生型同窝小鼠的更小、更短。这些结果表明xpg突变体小鼠的小脑浦肯野细胞有明显退化。通过对小脑皮质DNA片段化进行原位检测的研究表明,在突变小鼠的颗粒层中出现了一些脱氧核苷酸末端转移酶(TdT)介导的dUTP-生物素原位缺口标记(TUNEL)阳性细胞,但在浦肯野层中确认的细胞死亡很少。这些结果表明,突变体小鼠的小脑浦肯野细胞正在发生退化,其中尚未出现大量浦肯野细胞死亡,并且xpg缺陷小鼠中一些异常小脑症状的出现不仅是由于明显的浦肯野细胞退化,还由于其他细胞的损伤。