Physiological and pathological functions of a novel prion-like protein, PrPLP/Dpl.

新型朊病毒样蛋白 PrPLP/Dpl 的生理和病理功能。

• 批准号: 13470065
• 负责人: KATAMINE Shigeru
• 金额: $ 9.15万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470065/
• 关键词: prion diseases; prion protein; prion-like protein; gene-knockout mice; neurodegeneration; 海綿状変性

Certain lines of mice devoid of prion protein (PrP), such as Ngsk Prnp^<0/0>, exhibit late-onset ataxia due to Purkinje cell degeneration, which is rescued by a transgene encoding PrP. In these mice, PrP-like protein, PrPLP/Dpl, is ectopically expressed by neurons including Purkinje cells and glial cells. To explore mechanisms of the neurodegeneration, two types of transgenic (tg) mice expressing PrPLP/Dpl in all neurons, tg(N-PrPLPIDpI), or only Purkinje cells, tg(P-PrPLP/Dpl), were generated and subsequently backcrossed with non-ataxic Zrch I Prnp^<0/0> mice to eliminate the Prnp alleles. In contrast to the tg mice with the Prnp^<+/+> background, never showing neurological abnormalities, not only tg(N-PrPLP/Dpl) but also, tg(P-PrPLP/Dpl) mice with the Prnp^<0/0> alleles developed Purkinje cell degeneration after incubation periods inversely correlated to the expression levels of PrPLP/Dpl. Moreover, some of the tg mice hemizygous for Prnp allele (Prnp^<0+>) also succumbed to the dise … More ase but much later than those carrying the Prnp^<0/0> alleles. These results indicated that PrPLP/Dpl ectopically expressed by Purkinje cells itself executes a does-dependent deleterious effect on the cells, and that a stoichiometric antagonistic interaction between PrP and PrPLP/Dpl is crucially involved in the neurodegeneration. We next introduced five types of PrP transgenes including heterologous hamster and two mouse/hamster chimeric genes as well as two mutants, each of which encoded PrP lacking residues 23-88 (MHM2.del23-88) or with E199K substitution (Mo.E199K), into Ngsk Prnp^<0/0> mice. Only MHM2.de123-88 failed to rescue from the Purkinje cell death. Little difference was observed in pathology and onset of ataxia between Ngsk Prnp^<0/0> and MHM2.de123-88/Ngsk Prnp^<0/0>. No detergent-insoluble PrPLP/Dpl was detectable in the CNS of Ngsk Prnp^<0/0> even after the onset of ataxia. Our findings provide evidence that the N-terminal residues 23-88 of PrP containing the unique octapeptide-repeat region is crucial for preventing Purkinje cell death in the Prnp^<0/0> mice expressing PrPLP/Dpl in the neuron. Less

某些没有原始蛋白质（PRP）的小鼠（例如NGSK PRNP^<0/0>）是由于Purkinje细胞变性而暴露的晚期性共济失调，这是由编码PRP的转基因响应的。在这些小鼠中，PRP样蛋白PRPLP/DPL是由包括Purkinje细胞和神经胶质细胞在内的神经元表达的。为了探索神经退行性的机制，在所有神经元中表达PRPLP/DPL的两种类型的转基因（TG）小鼠，或仅生成非触发Zrch I Prnp^<0/0/0/0/0/0/0/0/0/0/purkinje细胞TG（PPRPLP/DPL）均产生，并随后将其全部反向。 In contrast to the tg mice with the Prnp^<+/+> background, never showing neurologic abnormalities, not only tg(N-PrPLP/Dpl) but also, tg(P-PrPLP/Dpl) mice with the Prnp^<0/0> alleles developed Purkinje cell degeneration after incubation periods inversely correlated to the expression levels of PrPLP/Dpl.此外，某些TG小鼠在PRNP等位基因（PRNP^<0+>）中也屈服于Disse…ASE…比携带PRNP^<0/0>等位基因的TG小鼠。这些结果表明，由Purkinje细胞本身以生态表达的PRPLP/DPL对细胞执行依赖性有害作用，并且PRP和PRPLP/DPL之间的化学计量拮抗相互作用完全参与神经变性。接下来，我们介绍了五种类型的PRP翻译，包括异源仓鼠和两个鼠标/仓鼠嵌合基因以及两个突变体，每个突变体编码的PRP缺乏保留23-88（MHM2.DEL23-88）或E199K替代（MO.E199K）（MO.E199K）（MO.E199K）（MO.E199K）（ngsk prnp prnp prnp prnp^<0/0/0> mice）。只有MHM2.DE123-88才能从Purkinje细胞死亡中救出。 NGSK PRNP^<0/0>和MHM2.DE123-88/ngsk PRNP^<0/0>之间的病理和共济失调的病理和共济失调的发作几乎没有差异。即使在共济失调开始后，也可以在NGSK PRNP^<0/0>的CNS中检测到无法确定不可差的PRPLP/DPL。我们的发现提供了证据表明，N端保留了23-88个含有独特的八肽肽重复区域的PRP，对于防止在神经元中表达PRPLP/DPL的PRNP^<0/0>小鼠中的Purkinje细胞死亡至关重要。较少的

项目成果

Lehmann S: "Ex vivo transmission of mouse-adapted prion strains to N2a and GT1-7 cell lines"Alzheimer's Disease : Advances in Etiology, Pathogenesis and Therapeutics. 679-686 (2001)

Lehmann S：“小鼠适应的朊病毒株离体传播至 N2a 和 GT1-7 细胞系”阿尔茨海默氏病：病因学、发病机制和治疗学的进展。

Atarashi, R., Sakaguchi, S., Shigematsu, K., Arima, K., Okimura, N., Yamaguchi, N., Li, A., Kopacek, J., Katamine, S: "Abnormal activation of glial cells in the brains of prion protein-deficient mice ctopically expressing prion protein-like protein, PrPLP

Atarashi, R.、Sakaguchi, S.、Shigematsu, K.、Arima, K.、Okimura, N.、Yamaguchi, N.、Li, A.、Kopacek, J.、Katamine, S：“神经胶质细胞的异常激活

Atarashi, R., Sakaguchi, S., Shigematsu, K Anima, K., Okimura, N., Yamaguchi, N.Li, A., Kopacek, J., Katamine, S.: "Abnormal activation of glial cells in the brains of prion protein-deficient mice ectopically expressing prion protein-like protein, PrPLP/D

Atarashi, R.、Sakaguchi, S.、Shigematsu、K Anima, K.、Okimura, N.、Yamaguchi, N.Li, A.、Kopacek, J.、Katamine, S.：“神经胶质细胞的异常激活

Yukitake M, Satoh J, Katamine S, Kuroda Y: "EAAT4 mRNA expression is preserved in the cerebellum of prion protein-deficient mice."Neurosci Lett. 352(3). 171-174 (2003)

Yukitake M、Satoh J、Katamine S、Kuroda Y：“EAAT4 mRNA 表达保留在朊病毒蛋白缺陷小鼠的小脑中。”Neurosci Lett。

Yukitake M, Satoh J, Katamine S, Kuroda Y: "EAAT4 mRNA expression is preserved in the cerebellum.of prion protein-deficient mice."Neurosci Lett.. 352(3). 171-174 (2003)

Yukitake M、Satoh J、Katamine S、Kuroda Y：“EAAT4 mRNA 表达保留在朊病毒蛋白缺陷小鼠的小脑中。”Neurosci Lett.. 352(3)。