Huntingtin-552片段在星形胶质细胞中的代谢及其对BDNF合成分泌的影响
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摘要
目的:构建亨廷顿舞蹈病(Huntington’s disease, HD)的体外原代星形胶质细胞模型;研究亨廷顿蛋白(Hungtingtin, htt)氨基端片段htt552在星形胶质细胞中的代谢途径及其对自噬水平的影响;研究htt552对星形胶质细胞脑源性神经营养因子(brain-derived neurotrophic factor , BDNF)转录与分泌的影响。
方法:采用腺病毒介导野生型与突变型htt552片段转染的方法构建HD的原代星形胶质细胞模型,利用Western Blot法和免疫荧光双标记法检测htt552在细胞内表达的水平及时程,采用MTT法测定腺病毒载体对细胞生存率的影响。运用Western Blot法及免疫荧光双标记法检测自噬抑制剂3-甲基腺嘌呤(3-MA)、自噬激活剂雷帕霉素(Rapamycin)和蛋白酶体抑制剂乳胞素(Lactacystin)对htt552在细胞内蓄积的影响。利用Western Blot法和Real-time PCR法检测细胞模型中自噬相关蛋白LC3II和Beclin1的表达情况。采用MDC法检测细胞自噬体形成的情况。采用ELISA和Western Blot法检测表达htt552后细胞内以及分泌的BDNF含量,采用Real-time PCR法测定htt552对细胞内BDNF mRNA及其不同转录本水平的影响。采用免疫荧光双标记法测定htt552与CREB结合蛋白(CBP)或高尔基复合体的共定位,从而研究htt552影响星形胶质细胞内BDNF转录和分泌的机制。
结果:腺病毒成功介导野生型与突变型htt552在原代星形胶质细胞中表达,该蛋白在细胞内主要分布于胞浆,而核内表达较少,部分细胞内可见突变型片段形成明显的聚集体。为了提高感染率和降低腺病毒对细胞的影响,以m.o.i为40的剂量加入病毒最为合适,感染率可达80%以上,感染后12hr直到7d都有蛋白表达,并且感染24hr到4d蛋白保持高表达。加入自噬抑制剂3-MA或者加入蛋白酶体抑制剂Lactacystin后,htt552在细胞内的蓄积量明显增加,其中突变型htt552蓄积更为显著且聚集体明显增多;加入自噬激活剂Rapamycin后,细胞内突变型htt552及其聚集体的含量显著降低,而对野生型片段无显著影响;在表达突变型和野生型htt552的两种细胞模型中,LC3II蛋白表达量显著增加,Beclin1的mRNA和蛋白表达水平均显著增高,MDC染色阳性的自噬囊泡数目显著增加;与野生型片段相比,表达突变型片段的细胞内LC3II与MDC染色阳性自噬囊泡的水平更高。免疫荧光实验显示,表达htt552-100Q的星形胶质细胞条件培养基(ACM)使原代培养的神经细胞突起分支数目减少、发育不良;ELISA和Western Blot法检测ACM中BDNF含量,结果显示htt552-100Q抑制BDNF的产生,而野生型片段无显著影响;Real-time PCR实验显示htt552-100Q抑制了细胞内BDNF mRNA水平,其原因和该突变片段显著抑制BDNF促进子II、III、IV有关,而野生型片段对BDNF的转录无显著影响;免疫荧光双标记法实验结果显示,htt552-100Q的可溶性片段及其聚集体和核内的CBP存在共定位,可能影响BDNF的转录过程;ELISA和Western Blot法检测细胞内BDNF含量,结果显示与表达野生型片段相比,表达htt552-100Q的细胞内BDNF含量显著升高,其中前体含量显著升高,而成熟的BDNF含量显著降低,提示htt552-100Q抑制BDNF的转运和成熟过程;免疫荧光双标记法实验显示,htt552-100Q的可溶性片段及其聚集体与BDNF囊泡和高尔基体共定位,并且使高尔基体的形态和分布发生变化,从而影响了BDNF的转运和分泌。
结论:成功构建高效表达htt552的体外HD星形胶质细胞模型,为研究HD的发病机制提供了一个新的平台;泛素-蛋白酶体途径和自噬/溶酶体途径均参与了htt552在细胞内的降解过程,而自噬-溶酶体途径是突变型htt552及其聚集体代谢的主要途径;htt552特别是突变型片段能够提高细胞自噬水平,从而加速自身代谢;突变型htt552能够抑制星形胶质细胞内BDNF的转录和分泌,从而对其周围神经元的生长发育造成影响;突变型htt552的可溶性片段及其聚集体与CBP、分泌囊泡及高尔基体结合,干扰了这些结构的正常功能,从而抑制BDNF转录和分泌。
Aim: To establish a glia Huntington’s disease (HD) model expressing wild-type and mutant N-terminal huntingtin fragment 1–552 amino acids (htt552), to study the contribution of autophagy/lysosomal pathway on the metabolism of the wild-type and mutant htt552 and to study the effects of wild-type and mutant htt552 on the production and secretion of brain-derived neurotrophic factor (BDNF) in astrocytes.
Method: In this study, we developed an in vitro model of HD by infecting primary cortical astrocytes of new-born rat with adenoviral vectors encoding the first 552 amino acids of wild-type (18Q) and mutant (100Q) htt. Western Blot analysis and double immunofluorescence (IF) were used to measure the levels and duration of htt552 expression in astrocytes. MTT assay was used to detect the viability of astrocytes after viral infection. The effects of autophagy specific inhibitor 3-methyadenine (3-MA), autophagy activator rapamycin, and the proteasome inhibitor lactacystin on htt552 accumulation were assessed in astrocytes expressing htt552 with Western Blot analysis and double immunofluorescence (IF). Proteins related to autophagy -- LC3 and Beclin1 were determined with Western Blot analysis and Real-time PCR. Autophagosomes were detected by monodansylcardeverine (MDC) immuofluorescence. Protein levels of BDNF in cell lysates and medium were detected by enzyme-linked immunosorbent assay (ELISA) and Western Blot analysis. The levels of BDNF mRNA and its four transcripts were detected with Real-time PCR to estimate the influence of htt552 on the transcription of BDNF. The co-location of htt552 with cAMP-response element binding protein (CBP) or Golgi complex was detected by double IF to estimate the mechanisms by which htt552 affects the transcription and secretion of BDNF.
Results: Wild-type and mutant htt552 transfected by adenovirus successfully expressed in primary astrocytes. They distributed predominantly in the cytoplasm with relatively low levels in the nucleus. Meanwhile, htt552-100Q produced the characteristic HD pathology by the appearance of cytoplasmic aggregates and intranuclear inclusions. To obtain high infection rate and low toxicity, the viral dose with an m.o.i. of 40 was optimal to our cell model, the duration of expression of htt552 lasted for about 7 days with a relatively high levels from 1 to 4 days post-infection. Treatments with the autophagy inhibitor (3-MA) or proteasome inhibitor (lactacystin) resulted in the augments of htt552 levels in cells, especially in those expressing mutant htt552. The activator of autophagy, rapamycin, significantly decreased the levels of mutant htt552 and the number of aggregates, however, had little effect on htt552-18Q accumulation. Western Blot analysis and Real time PCR revealed that the expression of LC3 and Beclin1 were up-regulated, and autophagosomes detected by MDC increased in cells expressing wild-type htt552 but it was more robust in the cells expressing mutant htt552. The result of IF assay shown that neurons cultured with astrocyte conditioned medium (ACM) from the cell expressing htt552-100Q exhibited shorter processes and fewer branchings than those with either ACM from the cell cultured normally or ACM from the cell expressing htt552-18Q. ELISA analysis revealed that total BDNF in the cell expressing htt552-100Q was increased compared with the cells expressing htt552-18Q. Western blot and ELISA analysis revealed that BDNF in ACM was down-regulated by htt552-100Q, and was not significantly changed by wild-type htt552. Western Blot assay further showed that proBDNF in astrocytes expressing with htt552-100Q was increased, but mature BDNF was decreased compared with the astrocytes expressing with htt552-18Q. Real time PCR revealed that BDNF mRNA levels were decreased in the astrocytes expressing htt552-100Q, as a result of the inhibition of promoter II、III、IV in BDNF gene. The result also showed that the levels of BDNF and its four transcripts were not significant change in the astrocytes expressing htt552-18Q. The result of double IF showed that htt552-100Q co-localized with CBP in the nucleus, which may have influence on the function of CBP. The result of double IF showed that htt552-100Q and its aggregates colocalized with BDNF puncta or Golgi complex, moreover, the morphology of Golgi complex and size of Golgi complex were changed by mutant htt552.
Conclusion: In vitro astrocytes HD model expressing wild-type and mutant N-terminal huntingtin fragment htt552 had been successfully established, which provided an advantageous system for research of HD pathogenesis in primary cortical astrocyte cultures. The ubiquitin-proteasome pathway (UPP) and autophagy/lysosomal pathway participated in the catabolism of htt552, and autophagy/lysosomal pathway was the major pathway in the catabolism of mutant htt552 and its aggregates. Wild-type and mutant htt552 fragments can both activate autophagy in cells and mutant fragments activated autophagy more robustly. Htt552-100Q can inhibit the transcription and secretion of BDNF in astrocytes, and then have influence on the development and function of neurons. Htt552-100Q and its aggregates may sequester CBP in the nucleus and BDNF vesicles in the cytoplasm. In addition, Htt552-100Q may have an aberrant effect on Golgi complex, thus disrupt the processing and secretion of BDNF.
方法:采用腺病毒介导野生型与突变型htt552片段转染的方法构建HD的原代星形胶质细胞模型,利用Western Blot法和免疫荧光双标记法检测htt552在细胞内表达的水平及时程,采用MTT法测定腺病毒载体对细胞生存率的影响。运用Western Blot法及免疫荧光双标记法检测自噬抑制剂3-甲基腺嘌呤(3-MA)、自噬激活剂雷帕霉素(Rapamycin)和蛋白酶体抑制剂乳胞素(Lactacystin)对htt552在细胞内蓄积的影响。利用Western Blot法和Real-time PCR法检测细胞模型中自噬相关蛋白LC3II和Beclin1的表达情况。采用MDC法检测细胞自噬体形成的情况。采用ELISA和Western Blot法检测表达htt552后细胞内以及分泌的BDNF含量,采用Real-time PCR法测定htt552对细胞内BDNF mRNA及其不同转录本水平的影响。采用免疫荧光双标记法测定htt552与CREB结合蛋白(CBP)或高尔基复合体的共定位,从而研究htt552影响星形胶质细胞内BDNF转录和分泌的机制。
结果:腺病毒成功介导野生型与突变型htt552在原代星形胶质细胞中表达,该蛋白在细胞内主要分布于胞浆,而核内表达较少,部分细胞内可见突变型片段形成明显的聚集体。为了提高感染率和降低腺病毒对细胞的影响,以m.o.i为40的剂量加入病毒最为合适,感染率可达80%以上,感染后12hr直到7d都有蛋白表达,并且感染24hr到4d蛋白保持高表达。加入自噬抑制剂3-MA或者加入蛋白酶体抑制剂Lactacystin后,htt552在细胞内的蓄积量明显增加,其中突变型htt552蓄积更为显著且聚集体明显增多;加入自噬激活剂Rapamycin后,细胞内突变型htt552及其聚集体的含量显著降低,而对野生型片段无显著影响;在表达突变型和野生型htt552的两种细胞模型中,LC3II蛋白表达量显著增加,Beclin1的mRNA和蛋白表达水平均显著增高,MDC染色阳性的自噬囊泡数目显著增加;与野生型片段相比,表达突变型片段的细胞内LC3II与MDC染色阳性自噬囊泡的水平更高。免疫荧光实验显示,表达htt552-100Q的星形胶质细胞条件培养基(ACM)使原代培养的神经细胞突起分支数目减少、发育不良;ELISA和Western Blot法检测ACM中BDNF含量,结果显示htt552-100Q抑制BDNF的产生,而野生型片段无显著影响;Real-time PCR实验显示htt552-100Q抑制了细胞内BDNF mRNA水平,其原因和该突变片段显著抑制BDNF促进子II、III、IV有关,而野生型片段对BDNF的转录无显著影响;免疫荧光双标记法实验结果显示,htt552-100Q的可溶性片段及其聚集体和核内的CBP存在共定位,可能影响BDNF的转录过程;ELISA和Western Blot法检测细胞内BDNF含量,结果显示与表达野生型片段相比,表达htt552-100Q的细胞内BDNF含量显著升高,其中前体含量显著升高,而成熟的BDNF含量显著降低,提示htt552-100Q抑制BDNF的转运和成熟过程;免疫荧光双标记法实验显示,htt552-100Q的可溶性片段及其聚集体与BDNF囊泡和高尔基体共定位,并且使高尔基体的形态和分布发生变化,从而影响了BDNF的转运和分泌。
结论:成功构建高效表达htt552的体外HD星形胶质细胞模型,为研究HD的发病机制提供了一个新的平台;泛素-蛋白酶体途径和自噬/溶酶体途径均参与了htt552在细胞内的降解过程,而自噬-溶酶体途径是突变型htt552及其聚集体代谢的主要途径;htt552特别是突变型片段能够提高细胞自噬水平,从而加速自身代谢;突变型htt552能够抑制星形胶质细胞内BDNF的转录和分泌,从而对其周围神经元的生长发育造成影响;突变型htt552的可溶性片段及其聚集体与CBP、分泌囊泡及高尔基体结合,干扰了这些结构的正常功能,从而抑制BDNF转录和分泌。
Aim: To establish a glia Huntington’s disease (HD) model expressing wild-type and mutant N-terminal huntingtin fragment 1–552 amino acids (htt552), to study the contribution of autophagy/lysosomal pathway on the metabolism of the wild-type and mutant htt552 and to study the effects of wild-type and mutant htt552 on the production and secretion of brain-derived neurotrophic factor (BDNF) in astrocytes.
Method: In this study, we developed an in vitro model of HD by infecting primary cortical astrocytes of new-born rat with adenoviral vectors encoding the first 552 amino acids of wild-type (18Q) and mutant (100Q) htt. Western Blot analysis and double immunofluorescence (IF) were used to measure the levels and duration of htt552 expression in astrocytes. MTT assay was used to detect the viability of astrocytes after viral infection. The effects of autophagy specific inhibitor 3-methyadenine (3-MA), autophagy activator rapamycin, and the proteasome inhibitor lactacystin on htt552 accumulation were assessed in astrocytes expressing htt552 with Western Blot analysis and double immunofluorescence (IF). Proteins related to autophagy -- LC3 and Beclin1 were determined with Western Blot analysis and Real-time PCR. Autophagosomes were detected by monodansylcardeverine (MDC) immuofluorescence. Protein levels of BDNF in cell lysates and medium were detected by enzyme-linked immunosorbent assay (ELISA) and Western Blot analysis. The levels of BDNF mRNA and its four transcripts were detected with Real-time PCR to estimate the influence of htt552 on the transcription of BDNF. The co-location of htt552 with cAMP-response element binding protein (CBP) or Golgi complex was detected by double IF to estimate the mechanisms by which htt552 affects the transcription and secretion of BDNF.
Results: Wild-type and mutant htt552 transfected by adenovirus successfully expressed in primary astrocytes. They distributed predominantly in the cytoplasm with relatively low levels in the nucleus. Meanwhile, htt552-100Q produced the characteristic HD pathology by the appearance of cytoplasmic aggregates and intranuclear inclusions. To obtain high infection rate and low toxicity, the viral dose with an m.o.i. of 40 was optimal to our cell model, the duration of expression of htt552 lasted for about 7 days with a relatively high levels from 1 to 4 days post-infection. Treatments with the autophagy inhibitor (3-MA) or proteasome inhibitor (lactacystin) resulted in the augments of htt552 levels in cells, especially in those expressing mutant htt552. The activator of autophagy, rapamycin, significantly decreased the levels of mutant htt552 and the number of aggregates, however, had little effect on htt552-18Q accumulation. Western Blot analysis and Real time PCR revealed that the expression of LC3 and Beclin1 were up-regulated, and autophagosomes detected by MDC increased in cells expressing wild-type htt552 but it was more robust in the cells expressing mutant htt552. The result of IF assay shown that neurons cultured with astrocyte conditioned medium (ACM) from the cell expressing htt552-100Q exhibited shorter processes and fewer branchings than those with either ACM from the cell cultured normally or ACM from the cell expressing htt552-18Q. ELISA analysis revealed that total BDNF in the cell expressing htt552-100Q was increased compared with the cells expressing htt552-18Q. Western blot and ELISA analysis revealed that BDNF in ACM was down-regulated by htt552-100Q, and was not significantly changed by wild-type htt552. Western Blot assay further showed that proBDNF in astrocytes expressing with htt552-100Q was increased, but mature BDNF was decreased compared with the astrocytes expressing with htt552-18Q. Real time PCR revealed that BDNF mRNA levels were decreased in the astrocytes expressing htt552-100Q, as a result of the inhibition of promoter II、III、IV in BDNF gene. The result also showed that the levels of BDNF and its four transcripts were not significant change in the astrocytes expressing htt552-18Q. The result of double IF showed that htt552-100Q co-localized with CBP in the nucleus, which may have influence on the function of CBP. The result of double IF showed that htt552-100Q and its aggregates colocalized with BDNF puncta or Golgi complex, moreover, the morphology of Golgi complex and size of Golgi complex were changed by mutant htt552.
Conclusion: In vitro astrocytes HD model expressing wild-type and mutant N-terminal huntingtin fragment htt552 had been successfully established, which provided an advantageous system for research of HD pathogenesis in primary cortical astrocyte cultures. The ubiquitin-proteasome pathway (UPP) and autophagy/lysosomal pathway participated in the catabolism of htt552, and autophagy/lysosomal pathway was the major pathway in the catabolism of mutant htt552 and its aggregates. Wild-type and mutant htt552 fragments can both activate autophagy in cells and mutant fragments activated autophagy more robustly. Htt552-100Q can inhibit the transcription and secretion of BDNF in astrocytes, and then have influence on the development and function of neurons. Htt552-100Q and its aggregates may sequester CBP in the nucleus and BDNF vesicles in the cytoplasm. In addition, Htt552-100Q may have an aberrant effect on Golgi complex, thus disrupt the processing and secretion of BDNF.
引文
1. Harper, P.S., et al. Hungtington’s Disease. 2ed. 1996: London: WB Saunder. 1-30.
2.李书林等.我国Hungtinton氏舞蹈病家系发病分析.中国优生与遗传杂志,2001,9(2):112-113.
3. Wang LH, et al. Treatment of Huntinton’s disease. Neurosi Bull, 2005, 21(3): 230-235.
4. The Huntington’s Disease Collaborative Research Group. A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington’s disease chromosomes. Cell 1993; 72: 971–983.
5. Gusella, J.F., et al. A polymorhic DNA marker genetically linked to Huntington’s disease. Nature, 1983, 306(5940): 234-238.
6. Perutz, M.F. Glutamine repeats and neurodegenerative disease: Molecular aspects. Trend Biol Sci, 1999, 24(2): 58-63.
7. Penney, J. B. et al. CAG repeat number governs the development rate of pathology in Huntington's disease. Ann. Neurol, 1997 , 41, 689-692.
8. DiFiglia, M., et al., Huntingtin is a cytoplasmic protein associated with vesicles in human and rat brain neurons. Neuron, 1995, 14(5): 1075-1081.
9. Gutekunst, C.A., et al. Identification and localization of huntingtin in brain and human lymphoblastoid cell lines with anti-fusion protein antibodies. Proc Natl Acad Sci USA, 1995, 92(19): 8710-8714.
10. Difiglia, M., et al., Aggregation of huntingtin in neuronal intranuclear inclusions and dystrophic neuritis in brain. Science, 1997, 277(5334): 1990-1993.
11. Davies, S.W., et al., Formation of neuronal intranuclear inclusions underlies the neurological dysfunction in mice transgenic for the HD mutation. Cell, 1997, 90(3): 537-548.
12. Saudou, F., et al. Huntingtin acts in the nucleus to induce apoptosis but death does not correlate with the formation of intranulear inclusions. Cell, 1998, 95(1): 55-56.
13. Qin ZH, Gu ZL. Huntingtin processing in pathogenesis of Huntington disease.ActaPharmacol Sin. 2004 Oct;25 (10):1243-9.
14. Zuccato C, et al. Loss of Huntingtin-mediated BDNF gene transcription in Huntington’s disease. Science. 2001 Jul 20;293(5529):493-498.
15. Gauthier LR, et al. Huntingtin controls neurotrophic support and survival of neurons by enhancing BDNF vesicular transport along microtubules. Cell. 2004 Jul 9;118(1):127-138.
16. Markiewicz I, et al. The role of astrocytes in the physiology and pathology of the central nervous system. Acta Neurobio Exp, 2006, 66 (4):343-358
17. Yongmei Chen, et al. Swanson. Astrocytes and Brain Injury. J Cereb Blood Flow Metab, 2003; 23 (2):137-148
18. Singhrao SK, et al.. Huntingtin protein colocalizes with lesions of neurodegenerative diseases: An investigation in Huntington’s, Alzheimer’s, and Pick’s disease. Exp Neurol. 1998 Apr;150 (2):213-22.
19. Hebb MO, et al. Expression of the Huntington’s disease gene is regulated in astrocytes in the arcuate nucleus of the hypothalamus of postpartum rats. FASEB J. 1999 Jun;13(9):1099-106.
20. Liévens JC, et al. Impaired glutamate uptake in the R6 Huntington’s disease transgenic mice. Neurobiol Dis. 2001 Oct;8(5):807-21.
21. Behrens PF, et al. Impaired glutamate transport and glutamate-glutamine cycling: downstream effects of the Huntington mutation. Brain. 2002 Aug;125(Pt 8):1908-22.
22. Shin JY, et al. Expression of mutant huntingtin in glial cells contributes to neuronal excitotoxicity. J Cell Biol. 2005 Dec 19;171(6):1001-1012.
23. Hershlo, A., et al. The ubiquitin system. Nature Med, 2000, 6: 1073-1081.
24. Pickart, C.M.. Mechanisms underlying ubiquitination. Annu Rev Biochem, 2001, 70: 503-533.
25. Doherty, F.J., et al. The ubiquitin-proteasome pathway of intracellular proteolysis. Essays Biochem, 2002, 38: 51-63.
26. Mizushima, N. The pleiotropic role of autophagy: from protein metabolism to bactericide. Cell Death Differ, 2005, 12: 1535-1541.
27. Doi, H., et al., Identification of ubiquitin-interacting proteins in purified polyglutamine aggregates. FEBS Lett, 2004, 571: 171-176.
28. Verhoef, L.G., et al., Aggregate formation inhibits proteasomal degradation of polyglutamine proteins. Hum Mol Genet, 2002, 11: 2689-2700.
29. Qin, Z.H., et al., Autophagy regulates the processing of amino terminal huntingtin fragments. Hum Mol Genet, 2003, 12(24): 3231-3244.
30. Kegel, K.B., et al., Huntingtin expression stimulates endosomal-lysosomal activity, endosome tubulation, and autophagy. Neurosci, 2000, 20: 7268-7278.
31. Michalik, A., et al. Pathogenesis of polyglutamine disorders: aggregation revisited. Hum Mol Genet, 2003, 12: R173-R186.
32. Benarroch EE. Neuron-astrocyte interactions: partnership for normal function and disease in the central nervous system. Mayo Clin Proc. 2005 Oct;80 (10):1326-38.
33. Wu H, et al. Differential regulation of neurotrophin expression in basal forebrain astrocytes by neuronal signals. J Neurosci Res. 2004 Apr 1;76(1):76-85.
34. Chou SY, et al. Expanded-polyglutamine huntingtin protein suppresses the secretion and production of a chemokine (CCL5/RANTES) by astrocytes. J Neurosci. 2008 Mar 26;28(13):3277-90.
35. Yuen EC, et al. Early BDNF, NT-3 , and NT-4 signalling events. Exp Neurol, 1999 , 159 ( 1 ) : 297-308.
36. Hansen MR, et al. Multiple distinct signal pathway, including an autocrine neurotrophic mechanism, contribute to the survival promoting effect of depolarization on spiral ganglion neurons in vivo. J Neurosci, 2001 , 21 ( 7 ) : 2256-2267.
37. Chiara Zuccato, et al. Role of brain-derived neurotrophic factor in Huntington’s disease. Progress in Neurobiology 81 (2007) 294–330.
38. Timmusk, T., et al. Multiple promoters direct tissue-specific expression of the rat BDNF gene. Neuron 1993, 10, 475–489.
39. Liu, Q.R., et al. Rodent BDNF genes, novel promoters, novel splice variants, and regulation by cocaine. Brain Res. 2006. 1067, 1–12.
40. Liu, Q.R., et al. Human brain derived neurotrophic factor (BDNF) genes, splicingpatterns, and assessments of associations with substance abuse and Parkinson’s Disease. Am. J. Med. Genet. B Neuropsychiatr. Genet. 2005.134, 93–103.
41. Hodgson, J.G., et al. A YAC mouse model for Huntington’s disease with full-length mutant huntingtin, cytoplasmic toxicity, and selective striatal neurodegeneration. Neuron 1999.23, 181–192.
42. Zuccato, C., et al. Progressive loss of BDNF in a mouse model of Huntington’s disease and rescue by BDNF delivery. Pharmacol. Res. 2005a. 52, 133–139.
43. Fusco, F.R., et al. Co-localization of brain-derived neurotrophic factor (BDNF) and wild-type huntingtin in normal and quinolinic acid-lesioned rat brain. Eur. J. Neurosci. 2003.18, 1093–1102.
44. Trushina, E., et al. Mutant huntingtin impairs axonal trafficking in mammalian neurons in vivo and in vitro. Mol. Cell. Biol. 2004.24, 8195–8209.
45. Gauthier, L.R., et al. Huntingtin controls neurotrophic support and survival of neurons by enhancing BDNF vesicular transport along microtubules. Cell 2004.118, 127–138.
46. McGeer, E.G. et al. Duplication of bio-chemical changes of Huntington's chorea by intrastriatal injections of glutamic and kainic acid. Nature, 1976, 263: 517-519.
47. Roberts, R., et al., Intrastriatal injections of quinolinic acid or kainic acid: differential patterns of cell survival and effects of data analysis on outcome. Exp. Neurol, 1993, 124: 274-282.
48. Guyot, M.C., et al., Quantifiable bradykinesia, gait abnormalities and Huntington's disease-like striatal lesions in rats chronically treated with 3-nitropropionic acid. Neuroscience, 1997, 79: 45-56.
49. Kirupa, S., et al., Transgenic models of Huntington’s disease. Phil. Trans. R. Soc. Lond. B, 1999, 354: 963-969.
50. Reddy, P.H., et al., Behavioural abnormalities and selective neuronal loss in HD transgenic mice expressing mutated full-length HD cDNA. Nat. Genet, 1998, 20: 198-202.
51. Hodgson, J.G., et al. AYAC mouse model for Huntington’s disease with full-length mutant huntingtin, cytoplasmic toxicity, and selective striatal neurodegeneration.Neuron, 1999, 23: 181-192.
52. Nasir, J., et al., Targeted disruption of the Huntington’s disease gene results in embryonic lethality and behavioral and morphological changes in heterozygotes. Cell, 1995, 81: 811-823.
53. White, J.K., et al., Huntingtin is required for neurogenesis and is not impaired by the Huntington’s disease CAG expansion. Nat. Genet, 1997, 17: 404-410.
54. Lin, C.H., et al. Neurological abnormalities in a knock-in mouse model of Huntington’s disease. Hum. Mol. Genet, 2001, 10: 137-144.
55. Almeida, L.P., et al., Lentiviral-mediated delivery of mutant huntingtin in the striatum of rats induces a selective neuropathology modulated by polyglutamine repeat size huntingtin expression levels and protein length. J Neurosci, 2002, 22: 3473-3483.
56. Zhen, L., et al., A putative drosophila homolog of the Huntington’s disease gene. Human Molecular Genetics, 1999, 8: 1807-1815.
57. Lawrence, M., et al. Fly models of Huntington’s disease. Human Molecular Genetics, 2003, 12: R187-R193.
58. Alex, P., et al., Expanded polyglutamines in Caenorhabditis elegans cause axonal abnormalities and severe dysfunction of PLM mechanosensory neurons without cell death, Proc Natl Acad Sci USA, 2001, 98: 13318-13323.
59. Moore, H., et al., Triplet repeats form secondary structures that escape DNA repair in yeast. Proc Natl Acad Sci USA, 1999, 96: 1504-1509.
60. Petersen A, et al. Expanded CAG repeats in exon 1 of the Huntington’s disease gene stimulate dopamine-mediated striatal neuron autophagy and degeneration. Hum Mol Genet 2001, 10: 1243–1254.
61. Hermel E, et al. Specific caspase interactions and amplification are involved in selective neuronal vulnerability in Huntington’s disease. Cell Death Differ 2004, 11: 424–438.
62. Zeron MM, et al. Potentiation of NMDA receptormediated excitotoxicity linked with intrinsic apoptotic pathway in YAC transgenic mouse model of Huntington’s disease. Mol Cell Neurosci 2004, 25: 469–479.
63. Qin ZH, et al. Huntingtin processing in pathogenesis of Huntington disease.Acta Pharmacol Sin. 2004 Oct;25 (10):1243-9.
64. Wellington CL, et al. Caspase cleavage of mutant huntingtin precedes neurodegeneration in Huntington's disease. J Neurosci. 2002 Sep 15;22(18):7862-72.
65. During MJ et al.. Towards gene therapy for the central nervous system. Mol Med Today 1998, 11: 485?493.
66. Kabeya, Y., et al., LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J, 2000, 19:5720-5728.
67. Ferrante RJ, et al. Selective sparing of a class of striatal neurons in Huntington’s disease. Science 1985; 230: 561–4.
68. Ferrante RJ, et al. Morphologic and histochemical characteristics of a spared subset of striatal neurons in Huntington’s disease. J Neuropathol Exp Neurol 1987; 46: 12–27.
69. Cicchetti F, et al. Striatal interneurons in Huntington’s disease: selective increase in the density of calretinin-immunoreactive medium-sized neurons. Mov Disord 1996; 11: 619–26
70. Lin-hui WANG, et al. Animal models of Huntington’s disease: implications in uncovering pathogenic mechanisms and developing therapies. Acta Pharmacologica Sinica 2006; 27 (10): 1287–1302.
71. Goldberg YP, et al. Cleavage of huntingtin by apopain, a proapoptotic cysteine protease, is modulated by the polyglutamine tract. Nat Genet 1996, 13: 442–449.
72. Wellington CL, et al. Caspase cleavage of gene products associated with triplet expansion disorders generates truncated fragments containing the polyglutamine tract. J Biol Chem 1998, 273: 9158–9167.
73. Wellington CL, et al. Inhibiting caspase cleavage of huntingtin reduces toxicity and aggregate formation in neuronal and non neuronal cells. J Biol Chem 2000, 275: 19831–19838.
74. Lin F, et al. Huntingtin cleavage induced by thrombin in vitro. Acta Biochim Biophys Sin 2007, 39: 15–18.
75. Wellington CL, et al. Caspase cleavage of mutant huntingtin precedes neurodegeneration in Huntington’s disease. J Neurosci 680 2002, 22: 7862–7872.
76. During MJ et al. Towards gene therapy for the central nervous system. Mol Med Today 1998, 11: 485?493.
77. Petersén A, et al.Expanded CAG repeats in exon 1 of the Huntington’s disease gene stimulate dopamine-mediated striatal neuron autophagy and degeneration. Hum Mol Genet 2001, 10: 1243?1254.
78. Saudou F, et al.. Huntingtin acts in the nucleus to induce apoptosis but death does not correlate with the formation of intranuclear inclusions. Cell 1998, 95: 55?66.
79. Zeron MM, et al. Potentiation of NMDA receptor-mediated excitotoxicity linked with intrinsic apoptotic pathway in YAC transgenic mouse model of Huntington’s disease. Mol Cell Neurosci 2004, 25: 469?479.
80. DiFiglia M, et al. Aggregation of huntingtin in neuronal intranuclear inclusions and dystrophic neurites in brain. Science 1997, 277: 1990–1993.
81. Sieradzan KA, et al. Huntington’s disease intranuclear inclusions contain truncated, ubiquitinated huntingtin protein. Exp Neurol 1999, 156: 92–99.
82. Qin ZH et al. Huntingtin processing in pathogenesis of Huntington disease. Acta Pharmacol Sin 2004, 25: 1243–1249.
83. Zala D, et al. Progressive and selective striatal degeneration in primary neuronal cultures using lentiviral vector coding for a mutant huntingtin fragment. Neurobiol Dis 2005, 20: 785?798.
84. Per, O.S., et al. 3-Methyladenine: specific inhibitor of autophagic/lysosomal protein degradation in isolated rat hepatocytes. Proc Natl Acad Sci USA, 1982, 79(6): 1889-1892.
85. Verhoef, L.G., et al., Aggregate formation inhibits proteasomal degradation of polyglutamine proteins. Hum Mol Genet, 2002, 11: 2689-2700.
86. Per, O.S., et al. 3-Methyladenine: specific inhibitor of autophagic/lysosomal protein degradation in isolated rat hepatocytes. Proc Natl Acad Sci USA, 1982, 79(6): 1889-1892.
87. Kova′cs, A.L., et al. Inhibition of autophagic sequestration and endogenous proteindegradation in isolated rat hepatocytes by methylated adenosine derivatives. FEBS Lett, 1981, 134: 194-196.
88. Klionsky, D.J., et al., Autophagy and P70S6 kinase. Autophagy, 2005, 1(1): 59-61.
89. Klionsky, D.J., et al., A unified nomenclature for yeast autophagy-related genes. Dev. Cell, 2003, 5(4): 539-545.
90. Lockshin, R.A., et al. Apoptosis, autophagy and more. Inter J Biochem Cell Biol, 2004, 36: 2405-2419.
91. Yashimori, T. Autophagy: a regulated bulk degradation process inside cells. Biochem Biophys Res Commun, 2004, 313(2): 453-458.
92. Liang, X.H., et al., Beclin 1 contains a leucine-rich nuclear export signal that is required for its autophagy and tumor suppressor function. Cancer Research 2001, 61(8): 3443-3449.
93. Yue, Z.Y., et al., Beclin 1, an autophagy gene essential for early embryonic development, is a haploinsufficient tumor suppressor. Proc Natl Acad Sci USA, 2003, 100(25): 15077-15082.
94. Teismann P, et al. Cellular pathology of Parkinson’s disease: astrocytes, microglia and inflammation. Cell Tissue Res, 2004; 318: 149-161
95. Timmusk, T., et al. Brain-derived neurotrophic factor expression in vivo is under the control of neuron-restrictive silencer element. J. Biol. Chem, 1999, 274, 1078–1084.
96. Zuccato, C., et al. Huntingtin interacts with REST/NRSF to modulate the transcription of NRSE-controlled neuronal genes. Nat. Genet., 2003, 35, 76–83.
97. Sugars, K.L, et al. Transcriptional abnormalities in Huntington disease. Trends Genet., 2003, 19, 233–238.
98. Sugars, K.L., et al. Decreased cAMP response element-mediated transcription: an early event in exon 1 and full-length cell models of Huntington’s disease that contributes to polyglutamine pathogenesis. J. Biol. Chem.,2004, 279, 4988–4999.
99. Steffan, J.S., et al. Histone deacetylase inhibitors arrest polyglutamine-dependent neurodegeneration in Drosophila. Nature , 2001, 413, 739–743.
100. Yu, Z.X., et al. Huntingtin inclusions do not deplete polyglutamine-containing transcription factors in HD mice. Hum.Mol. Genet, 2002, 11, 905–914.
101. Gines, S., et al. Specific progressive camp reduction implicates energy deficit in presymptomatic Huntington’s disease knock-in mice. Hum. Mol. Genet, 2003, 12, 497–508.
102. Giampa, C., et al. Striatal modulation of cAMPresponse- element-binding protein (CREB) after excitotoxic lesions: implications with neuronal vulnerability in Huntington’s disease. Eur. J. Neurosci., 2006, 23, 11–20.
103. Dunah, A.W., et al. Sp1 and TAFII130 transcriptional activity disrupted in early Huntington’s disease. Science , 2002., 296, 2238–2243.
104. Zuccato, C., et al. Progressive loss of BDNF in a mouse model of Huntington’s disease and rescue by BDNF delivery. Pharmacol. Res, 2005,.52, 133–139.
105. Takeuchi, Y., et al. Analysis on the promoter region of exon IV brain-derived neurotrophic factor in NG108-15 cells. J. Neurochem, 2002, 83, 67–79.
106. Li, S.H., et al. Interaction of Huntington disease protein with transcriptional activator Sp1. Mol. Cell. Biol., 2002., 22, 1277–1287.
107. Li, J.Y., et al. Huntington’s disease: a synaptopathy? Trends Mol. Med, 2003., 9, 414–420.
108. Fusco, F.R., et al. Co-localization of brain-derived neuro-trophic factor (BDNF) and wild-type huntingtin in normal and quinolinic acid-lesioned rat brain. Eur. J. Neurosci, 2003, 18, 1093–1102.
109. Block-Galarza, J. et al. Fast transport and retrograde movement of huntingtin and HAP 1 in axons. Neuroreport ,1997, 8, 2247–2251.
110. Engelender, S., et al. Huntingtin-associated protein 1 (HAP1) interacts with the p150Glued subunit of dynactin. Hum. Mol.Genet, 1997, 6, 2205–2212
111. McGuire, J.R., et al. Interaction of Huntingtinassociated protein-1 with kinesin light chain: implications in intracellular trafficking in neurons. J. Biol. Chem, 2005, 281, 3552–3559.
112. Borrell-Pages, M., et al. Cystamine and cysteamine increase brain levels of BDNF in Huntington disease via HSJ1b and transglutaminase. J. Clin. Invest,2006, 116, 1410–1424.
113. Cheetham, M.E., et al. Inhibition of hsc70- catalysed clathrin uncoating by HSJ1 proteins. Biochem, 1996, J. 319, 103–108.
114. Gleeson, P.A., et al. Domains of the TGN: coats, tethers and G proteins. Traffic, 2004, 5, 315–326.
115. Canals, J.M., et al. Brain-derived neurotrophic factor regulates the onset and severity of motor dysfunction associated with enkephalinergic neuronal degeneration in Huntington’s disease. J. Neurosci, 2004, 24, 7727–7739.
116. Mangiarini, L., et al. Exon 1 of the HD gene with an expanded CAG repeat is sufficient to cause a progressive neurological phenotype in transgenic mice. Cell,1996, 87, 493–506.
117. Xu, B, et al. Slowing progression of Huntington’s disease by BDNF overexpression. In: Proceedings of the 36th Annual Congress of the Society for Neuroscience (abstract, 2006., 277.17/MM3).
118. Tardito, D et al. Signaling pathways regulating gene expression, neuroplasticity, and neurotrophic mechanisms in the action of antidepressants: a critical overview. Pharmacol. Rev, 2006, 58, 115–134.
119. Chuang, D.M.. Neuroprotective and neurotrophic actions of the mood stabilizer lithium: can it be used to treat neurodegenerative diseases? Crit. Rev. Neurobiol., 2004,16, 83–90.
120. Katoh-Semba, R., et al. Riluzole enhances expression of brain-derived neurotrophic factor with consequent proliferation of granule precursor cells in the rat hippocampus. FASEB J, 2002, 6, 1328–1330.
121. Marvanova, M., et al. The neuroprotective agent memantine induces brain-derived neurotrophic factor and trkB receptor expression in rat brain. Mol. Cell. Neurosci, 2001, 18, 247–258.
2.李书林等.我国Hungtinton氏舞蹈病家系发病分析.中国优生与遗传杂志,2001,9(2):112-113.
3. Wang LH, et al. Treatment of Huntinton’s disease. Neurosi Bull, 2005, 21(3): 230-235.
4. The Huntington’s Disease Collaborative Research Group. A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington’s disease chromosomes. Cell 1993; 72: 971–983.
5. Gusella, J.F., et al. A polymorhic DNA marker genetically linked to Huntington’s disease. Nature, 1983, 306(5940): 234-238.
6. Perutz, M.F. Glutamine repeats and neurodegenerative disease: Molecular aspects. Trend Biol Sci, 1999, 24(2): 58-63.
7. Penney, J. B. et al. CAG repeat number governs the development rate of pathology in Huntington's disease. Ann. Neurol, 1997 , 41, 689-692.
8. DiFiglia, M., et al., Huntingtin is a cytoplasmic protein associated with vesicles in human and rat brain neurons. Neuron, 1995, 14(5): 1075-1081.
9. Gutekunst, C.A., et al. Identification and localization of huntingtin in brain and human lymphoblastoid cell lines with anti-fusion protein antibodies. Proc Natl Acad Sci USA, 1995, 92(19): 8710-8714.
10. Difiglia, M., et al., Aggregation of huntingtin in neuronal intranuclear inclusions and dystrophic neuritis in brain. Science, 1997, 277(5334): 1990-1993.
11. Davies, S.W., et al., Formation of neuronal intranuclear inclusions underlies the neurological dysfunction in mice transgenic for the HD mutation. Cell, 1997, 90(3): 537-548.
12. Saudou, F., et al. Huntingtin acts in the nucleus to induce apoptosis but death does not correlate with the formation of intranulear inclusions. Cell, 1998, 95(1): 55-56.
13. Qin ZH, Gu ZL. Huntingtin processing in pathogenesis of Huntington disease.ActaPharmacol Sin. 2004 Oct;25 (10):1243-9.
14. Zuccato C, et al. Loss of Huntingtin-mediated BDNF gene transcription in Huntington’s disease. Science. 2001 Jul 20;293(5529):493-498.
15. Gauthier LR, et al. Huntingtin controls neurotrophic support and survival of neurons by enhancing BDNF vesicular transport along microtubules. Cell. 2004 Jul 9;118(1):127-138.
16. Markiewicz I, et al. The role of astrocytes in the physiology and pathology of the central nervous system. Acta Neurobio Exp, 2006, 66 (4):343-358
17. Yongmei Chen, et al. Swanson. Astrocytes and Brain Injury. J Cereb Blood Flow Metab, 2003; 23 (2):137-148
18. Singhrao SK, et al.. Huntingtin protein colocalizes with lesions of neurodegenerative diseases: An investigation in Huntington’s, Alzheimer’s, and Pick’s disease. Exp Neurol. 1998 Apr;150 (2):213-22.
19. Hebb MO, et al. Expression of the Huntington’s disease gene is regulated in astrocytes in the arcuate nucleus of the hypothalamus of postpartum rats. FASEB J. 1999 Jun;13(9):1099-106.
20. Liévens JC, et al. Impaired glutamate uptake in the R6 Huntington’s disease transgenic mice. Neurobiol Dis. 2001 Oct;8(5):807-21.
21. Behrens PF, et al. Impaired glutamate transport and glutamate-glutamine cycling: downstream effects of the Huntington mutation. Brain. 2002 Aug;125(Pt 8):1908-22.
22. Shin JY, et al. Expression of mutant huntingtin in glial cells contributes to neuronal excitotoxicity. J Cell Biol. 2005 Dec 19;171(6):1001-1012.
23. Hershlo, A., et al. The ubiquitin system. Nature Med, 2000, 6: 1073-1081.
24. Pickart, C.M.. Mechanisms underlying ubiquitination. Annu Rev Biochem, 2001, 70: 503-533.
25. Doherty, F.J., et al. The ubiquitin-proteasome pathway of intracellular proteolysis. Essays Biochem, 2002, 38: 51-63.
26. Mizushima, N. The pleiotropic role of autophagy: from protein metabolism to bactericide. Cell Death Differ, 2005, 12: 1535-1541.
27. Doi, H., et al., Identification of ubiquitin-interacting proteins in purified polyglutamine aggregates. FEBS Lett, 2004, 571: 171-176.
28. Verhoef, L.G., et al., Aggregate formation inhibits proteasomal degradation of polyglutamine proteins. Hum Mol Genet, 2002, 11: 2689-2700.
29. Qin, Z.H., et al., Autophagy regulates the processing of amino terminal huntingtin fragments. Hum Mol Genet, 2003, 12(24): 3231-3244.
30. Kegel, K.B., et al., Huntingtin expression stimulates endosomal-lysosomal activity, endosome tubulation, and autophagy. Neurosci, 2000, 20: 7268-7278.
31. Michalik, A., et al. Pathogenesis of polyglutamine disorders: aggregation revisited. Hum Mol Genet, 2003, 12: R173-R186.
32. Benarroch EE. Neuron-astrocyte interactions: partnership for normal function and disease in the central nervous system. Mayo Clin Proc. 2005 Oct;80 (10):1326-38.
33. Wu H, et al. Differential regulation of neurotrophin expression in basal forebrain astrocytes by neuronal signals. J Neurosci Res. 2004 Apr 1;76(1):76-85.
34. Chou SY, et al. Expanded-polyglutamine huntingtin protein suppresses the secretion and production of a chemokine (CCL5/RANTES) by astrocytes. J Neurosci. 2008 Mar 26;28(13):3277-90.
35. Yuen EC, et al. Early BDNF, NT-3 , and NT-4 signalling events. Exp Neurol, 1999 , 159 ( 1 ) : 297-308.
36. Hansen MR, et al. Multiple distinct signal pathway, including an autocrine neurotrophic mechanism, contribute to the survival promoting effect of depolarization on spiral ganglion neurons in vivo. J Neurosci, 2001 , 21 ( 7 ) : 2256-2267.
37. Chiara Zuccato, et al. Role of brain-derived neurotrophic factor in Huntington’s disease. Progress in Neurobiology 81 (2007) 294–330.
38. Timmusk, T., et al. Multiple promoters direct tissue-specific expression of the rat BDNF gene. Neuron 1993, 10, 475–489.
39. Liu, Q.R., et al. Rodent BDNF genes, novel promoters, novel splice variants, and regulation by cocaine. Brain Res. 2006. 1067, 1–12.
40. Liu, Q.R., et al. Human brain derived neurotrophic factor (BDNF) genes, splicingpatterns, and assessments of associations with substance abuse and Parkinson’s Disease. Am. J. Med. Genet. B Neuropsychiatr. Genet. 2005.134, 93–103.
41. Hodgson, J.G., et al. A YAC mouse model for Huntington’s disease with full-length mutant huntingtin, cytoplasmic toxicity, and selective striatal neurodegeneration. Neuron 1999.23, 181–192.
42. Zuccato, C., et al. Progressive loss of BDNF in a mouse model of Huntington’s disease and rescue by BDNF delivery. Pharmacol. Res. 2005a. 52, 133–139.
43. Fusco, F.R., et al. Co-localization of brain-derived neurotrophic factor (BDNF) and wild-type huntingtin in normal and quinolinic acid-lesioned rat brain. Eur. J. Neurosci. 2003.18, 1093–1102.
44. Trushina, E., et al. Mutant huntingtin impairs axonal trafficking in mammalian neurons in vivo and in vitro. Mol. Cell. Biol. 2004.24, 8195–8209.
45. Gauthier, L.R., et al. Huntingtin controls neurotrophic support and survival of neurons by enhancing BDNF vesicular transport along microtubules. Cell 2004.118, 127–138.
46. McGeer, E.G. et al. Duplication of bio-chemical changes of Huntington's chorea by intrastriatal injections of glutamic and kainic acid. Nature, 1976, 263: 517-519.
47. Roberts, R., et al., Intrastriatal injections of quinolinic acid or kainic acid: differential patterns of cell survival and effects of data analysis on outcome. Exp. Neurol, 1993, 124: 274-282.
48. Guyot, M.C., et al., Quantifiable bradykinesia, gait abnormalities and Huntington's disease-like striatal lesions in rats chronically treated with 3-nitropropionic acid. Neuroscience, 1997, 79: 45-56.
49. Kirupa, S., et al., Transgenic models of Huntington’s disease. Phil. Trans. R. Soc. Lond. B, 1999, 354: 963-969.
50. Reddy, P.H., et al., Behavioural abnormalities and selective neuronal loss in HD transgenic mice expressing mutated full-length HD cDNA. Nat. Genet, 1998, 20: 198-202.
51. Hodgson, J.G., et al. AYAC mouse model for Huntington’s disease with full-length mutant huntingtin, cytoplasmic toxicity, and selective striatal neurodegeneration.Neuron, 1999, 23: 181-192.
52. Nasir, J., et al., Targeted disruption of the Huntington’s disease gene results in embryonic lethality and behavioral and morphological changes in heterozygotes. Cell, 1995, 81: 811-823.
53. White, J.K., et al., Huntingtin is required for neurogenesis and is not impaired by the Huntington’s disease CAG expansion. Nat. Genet, 1997, 17: 404-410.
54. Lin, C.H., et al. Neurological abnormalities in a knock-in mouse model of Huntington’s disease. Hum. Mol. Genet, 2001, 10: 137-144.
55. Almeida, L.P., et al., Lentiviral-mediated delivery of mutant huntingtin in the striatum of rats induces a selective neuropathology modulated by polyglutamine repeat size huntingtin expression levels and protein length. J Neurosci, 2002, 22: 3473-3483.
56. Zhen, L., et al., A putative drosophila homolog of the Huntington’s disease gene. Human Molecular Genetics, 1999, 8: 1807-1815.
57. Lawrence, M., et al. Fly models of Huntington’s disease. Human Molecular Genetics, 2003, 12: R187-R193.
58. Alex, P., et al., Expanded polyglutamines in Caenorhabditis elegans cause axonal abnormalities and severe dysfunction of PLM mechanosensory neurons without cell death, Proc Natl Acad Sci USA, 2001, 98: 13318-13323.
59. Moore, H., et al., Triplet repeats form secondary structures that escape DNA repair in yeast. Proc Natl Acad Sci USA, 1999, 96: 1504-1509.
60. Petersen A, et al. Expanded CAG repeats in exon 1 of the Huntington’s disease gene stimulate dopamine-mediated striatal neuron autophagy and degeneration. Hum Mol Genet 2001, 10: 1243–1254.
61. Hermel E, et al. Specific caspase interactions and amplification are involved in selective neuronal vulnerability in Huntington’s disease. Cell Death Differ 2004, 11: 424–438.
62. Zeron MM, et al. Potentiation of NMDA receptormediated excitotoxicity linked with intrinsic apoptotic pathway in YAC transgenic mouse model of Huntington’s disease. Mol Cell Neurosci 2004, 25: 469–479.
63. Qin ZH, et al. Huntingtin processing in pathogenesis of Huntington disease.Acta Pharmacol Sin. 2004 Oct;25 (10):1243-9.
64. Wellington CL, et al. Caspase cleavage of mutant huntingtin precedes neurodegeneration in Huntington's disease. J Neurosci. 2002 Sep 15;22(18):7862-72.
65. During MJ et al.. Towards gene therapy for the central nervous system. Mol Med Today 1998, 11: 485?493.
66. Kabeya, Y., et al., LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J, 2000, 19:5720-5728.
67. Ferrante RJ, et al. Selective sparing of a class of striatal neurons in Huntington’s disease. Science 1985; 230: 561–4.
68. Ferrante RJ, et al. Morphologic and histochemical characteristics of a spared subset of striatal neurons in Huntington’s disease. J Neuropathol Exp Neurol 1987; 46: 12–27.
69. Cicchetti F, et al. Striatal interneurons in Huntington’s disease: selective increase in the density of calretinin-immunoreactive medium-sized neurons. Mov Disord 1996; 11: 619–26
70. Lin-hui WANG, et al. Animal models of Huntington’s disease: implications in uncovering pathogenic mechanisms and developing therapies. Acta Pharmacologica Sinica 2006; 27 (10): 1287–1302.
71. Goldberg YP, et al. Cleavage of huntingtin by apopain, a proapoptotic cysteine protease, is modulated by the polyglutamine tract. Nat Genet 1996, 13: 442–449.
72. Wellington CL, et al. Caspase cleavage of gene products associated with triplet expansion disorders generates truncated fragments containing the polyglutamine tract. J Biol Chem 1998, 273: 9158–9167.
73. Wellington CL, et al. Inhibiting caspase cleavage of huntingtin reduces toxicity and aggregate formation in neuronal and non neuronal cells. J Biol Chem 2000, 275: 19831–19838.
74. Lin F, et al. Huntingtin cleavage induced by thrombin in vitro. Acta Biochim Biophys Sin 2007, 39: 15–18.
75. Wellington CL, et al. Caspase cleavage of mutant huntingtin precedes neurodegeneration in Huntington’s disease. J Neurosci 680 2002, 22: 7862–7872.
76. During MJ et al. Towards gene therapy for the central nervous system. Mol Med Today 1998, 11: 485?493.
77. Petersén A, et al.Expanded CAG repeats in exon 1 of the Huntington’s disease gene stimulate dopamine-mediated striatal neuron autophagy and degeneration. Hum Mol Genet 2001, 10: 1243?1254.
78. Saudou F, et al.. Huntingtin acts in the nucleus to induce apoptosis but death does not correlate with the formation of intranuclear inclusions. Cell 1998, 95: 55?66.
79. Zeron MM, et al. Potentiation of NMDA receptor-mediated excitotoxicity linked with intrinsic apoptotic pathway in YAC transgenic mouse model of Huntington’s disease. Mol Cell Neurosci 2004, 25: 469?479.
80. DiFiglia M, et al. Aggregation of huntingtin in neuronal intranuclear inclusions and dystrophic neurites in brain. Science 1997, 277: 1990–1993.
81. Sieradzan KA, et al. Huntington’s disease intranuclear inclusions contain truncated, ubiquitinated huntingtin protein. Exp Neurol 1999, 156: 92–99.
82. Qin ZH et al. Huntingtin processing in pathogenesis of Huntington disease. Acta Pharmacol Sin 2004, 25: 1243–1249.
83. Zala D, et al. Progressive and selective striatal degeneration in primary neuronal cultures using lentiviral vector coding for a mutant huntingtin fragment. Neurobiol Dis 2005, 20: 785?798.
84. Per, O.S., et al. 3-Methyladenine: specific inhibitor of autophagic/lysosomal protein degradation in isolated rat hepatocytes. Proc Natl Acad Sci USA, 1982, 79(6): 1889-1892.
85. Verhoef, L.G., et al., Aggregate formation inhibits proteasomal degradation of polyglutamine proteins. Hum Mol Genet, 2002, 11: 2689-2700.
86. Per, O.S., et al. 3-Methyladenine: specific inhibitor of autophagic/lysosomal protein degradation in isolated rat hepatocytes. Proc Natl Acad Sci USA, 1982, 79(6): 1889-1892.
87. Kova′cs, A.L., et al. Inhibition of autophagic sequestration and endogenous proteindegradation in isolated rat hepatocytes by methylated adenosine derivatives. FEBS Lett, 1981, 134: 194-196.
88. Klionsky, D.J., et al., Autophagy and P70S6 kinase. Autophagy, 2005, 1(1): 59-61.
89. Klionsky, D.J., et al., A unified nomenclature for yeast autophagy-related genes. Dev. Cell, 2003, 5(4): 539-545.
90. Lockshin, R.A., et al. Apoptosis, autophagy and more. Inter J Biochem Cell Biol, 2004, 36: 2405-2419.
91. Yashimori, T. Autophagy: a regulated bulk degradation process inside cells. Biochem Biophys Res Commun, 2004, 313(2): 453-458.
92. Liang, X.H., et al., Beclin 1 contains a leucine-rich nuclear export signal that is required for its autophagy and tumor suppressor function. Cancer Research 2001, 61(8): 3443-3449.
93. Yue, Z.Y., et al., Beclin 1, an autophagy gene essential for early embryonic development, is a haploinsufficient tumor suppressor. Proc Natl Acad Sci USA, 2003, 100(25): 15077-15082.
94. Teismann P, et al. Cellular pathology of Parkinson’s disease: astrocytes, microglia and inflammation. Cell Tissue Res, 2004; 318: 149-161
95. Timmusk, T., et al. Brain-derived neurotrophic factor expression in vivo is under the control of neuron-restrictive silencer element. J. Biol. Chem, 1999, 274, 1078–1084.
96. Zuccato, C., et al. Huntingtin interacts with REST/NRSF to modulate the transcription of NRSE-controlled neuronal genes. Nat. Genet., 2003, 35, 76–83.
97. Sugars, K.L, et al. Transcriptional abnormalities in Huntington disease. Trends Genet., 2003, 19, 233–238.
98. Sugars, K.L., et al. Decreased cAMP response element-mediated transcription: an early event in exon 1 and full-length cell models of Huntington’s disease that contributes to polyglutamine pathogenesis. J. Biol. Chem.,2004, 279, 4988–4999.
99. Steffan, J.S., et al. Histone deacetylase inhibitors arrest polyglutamine-dependent neurodegeneration in Drosophila. Nature , 2001, 413, 739–743.
100. Yu, Z.X., et al. Huntingtin inclusions do not deplete polyglutamine-containing transcription factors in HD mice. Hum.Mol. Genet, 2002, 11, 905–914.
101. Gines, S., et al. Specific progressive camp reduction implicates energy deficit in presymptomatic Huntington’s disease knock-in mice. Hum. Mol. Genet, 2003, 12, 497–508.
102. Giampa, C., et al. Striatal modulation of cAMPresponse- element-binding protein (CREB) after excitotoxic lesions: implications with neuronal vulnerability in Huntington’s disease. Eur. J. Neurosci., 2006, 23, 11–20.
103. Dunah, A.W., et al. Sp1 and TAFII130 transcriptional activity disrupted in early Huntington’s disease. Science , 2002., 296, 2238–2243.
104. Zuccato, C., et al. Progressive loss of BDNF in a mouse model of Huntington’s disease and rescue by BDNF delivery. Pharmacol. Res, 2005,.52, 133–139.
105. Takeuchi, Y., et al. Analysis on the promoter region of exon IV brain-derived neurotrophic factor in NG108-15 cells. J. Neurochem, 2002, 83, 67–79.
106. Li, S.H., et al. Interaction of Huntington disease protein with transcriptional activator Sp1. Mol. Cell. Biol., 2002., 22, 1277–1287.
107. Li, J.Y., et al. Huntington’s disease: a synaptopathy? Trends Mol. Med, 2003., 9, 414–420.
108. Fusco, F.R., et al. Co-localization of brain-derived neuro-trophic factor (BDNF) and wild-type huntingtin in normal and quinolinic acid-lesioned rat brain. Eur. J. Neurosci, 2003, 18, 1093–1102.
109. Block-Galarza, J. et al. Fast transport and retrograde movement of huntingtin and HAP 1 in axons. Neuroreport ,1997, 8, 2247–2251.
110. Engelender, S., et al. Huntingtin-associated protein 1 (HAP1) interacts with the p150Glued subunit of dynactin. Hum. Mol.Genet, 1997, 6, 2205–2212
111. McGuire, J.R., et al. Interaction of Huntingtinassociated protein-1 with kinesin light chain: implications in intracellular trafficking in neurons. J. Biol. Chem, 2005, 281, 3552–3559.
112. Borrell-Pages, M., et al. Cystamine and cysteamine increase brain levels of BDNF in Huntington disease via HSJ1b and transglutaminase. J. Clin. Invest,2006, 116, 1410–1424.
113. Cheetham, M.E., et al. Inhibition of hsc70- catalysed clathrin uncoating by HSJ1 proteins. Biochem, 1996, J. 319, 103–108.
114. Gleeson, P.A., et al. Domains of the TGN: coats, tethers and G proteins. Traffic, 2004, 5, 315–326.
115. Canals, J.M., et al. Brain-derived neurotrophic factor regulates the onset and severity of motor dysfunction associated with enkephalinergic neuronal degeneration in Huntington’s disease. J. Neurosci, 2004, 24, 7727–7739.
116. Mangiarini, L., et al. Exon 1 of the HD gene with an expanded CAG repeat is sufficient to cause a progressive neurological phenotype in transgenic mice. Cell,1996, 87, 493–506.
117. Xu, B, et al. Slowing progression of Huntington’s disease by BDNF overexpression. In: Proceedings of the 36th Annual Congress of the Society for Neuroscience (abstract, 2006., 277.17/MM3).
118. Tardito, D et al. Signaling pathways regulating gene expression, neuroplasticity, and neurotrophic mechanisms in the action of antidepressants: a critical overview. Pharmacol. Rev, 2006, 58, 115–134.
119. Chuang, D.M.. Neuroprotective and neurotrophic actions of the mood stabilizer lithium: can it be used to treat neurodegenerative diseases? Crit. Rev. Neurobiol., 2004,16, 83–90.
120. Katoh-Semba, R., et al. Riluzole enhances expression of brain-derived neurotrophic factor with consequent proliferation of granule precursor cells in the rat hippocampus. FASEB J, 2002, 6, 1328–1330.
121. Marvanova, M., et al. The neuroprotective agent memantine induces brain-derived neurotrophic factor and trkB receptor expression in rat brain. Mol. Cell. Neurosci, 2001, 18, 247–258.