基因

藏刀九段

Novel splicing variant of the human orphan nuclear receptor Nurr1 gene

XU Ping-yi 徐评议 and LE Wei-dong 乐卫东

Keywords: splicing site · exon5 · Nurr1 gene

Department of Neurology, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China (Xu PY) Health and Science Center, Shanghai Institute for Biological Sciences, Ruijin Hospital, Shanghai 200025, China (Le WD) Correspondence t Le Wei-dong, M.D., Room 1209, Science and Education Building, Institute of Biomedical Sciences, Ruijin Hospital, Shanghai 2nd Medical University, 197 Ruijin Second Road, Shanghai 200025, China (Tel: 86-21-64677503. Fax: 86-21-64670637. Email: [email protected]) This study was supported by a grant from Chinese National Science Foundation (No. 30370509), Chinese Oversea Scholar Foundation (No. 2003-406), the 100-Talent Project, sponsored by the Chinese Academy of Sciences (No. 2002298), and by a research grant from the Health and Sciences Center, Shanghai Institute for Biological Sciences (No. J0041-1402).

Background Nurr1 is a member of the nuclear receptor superfamily of transcription factors. The objective of the present study was to identify novel splicing variants of the gene in neuronal and non-neuronal tissues and determine their functions. Methods Reverse transcription-polymerase chain reaction (RT-PCR) analysis was used to screen for Nurr1 splice variants in the adult human central nervous system (CNS) and in other tissues such as lymphocytes, and liver, muscle, and kidney cells. Functional assays of the variants were performed by measuring Nurr1 response element (NuRE) transcriptional activity in vitro. Results In this study, the authors identified a novel splicing variant of Nurr1 within exon 5, found in multiple adult human tissues, including lymphocytes, and liver, muscle, and kidney cells, but not in the brain or spinal cord. Sequencing analysis showed the variant has a 75 bp deletion between nucleotides 1402 and 1476. A functional assay of the Nurr1-c splicing variant, performed by measuring NuRE transcriptional activity in vitro, detected a 39% lower level of luciferase (LUC) activity (P<0.05). Conclusion A novel splicing variant of Nurr1 exists in human non-neuronal tissues and functional assays suggest that the variant may act as an alternate transcription regulator.

Chin Med J 2004; 117(6):899-902

The orphan nuclear receptor, Nurr1, is a member of the steroid/thyroid hormone nuclear receptor superfamily. The gene has been mapped to chromosome 2q22-23, and contains eight exons and seven introns, with a total size of 9.822 kb.［1］ It has been demonstrated that Nurr1 is not only essential for final differentiation of ventral mesencephalic dopaminergic (DAergic) precursor neurons into the mature DAergic phenotype, but is also required for the normal function of mature DAergic neurons.［2,3］ Several studies have suggested that the Nurr1 gene may be involved in Parkinson’s disease, schizophrenia, and manic-depressive disorder.［4,5］ To date, several variants of Nurr1 with N-terminal and C-terminal domains of variable length have been isolated from human and mouse fetal brain tissues and lymphocytes. Most of these splice variants of Nurr1 are within exon 7.［6,7］ METHODS

Total RNA was isolated from adult human peripheral blood lymphocytes and postmortem brain and spinal cord tissue using the simian virus (SV) total RNA isolation system (Promega, Madison, USA). Total RNA from the adult human kidney, liver, and skeletal muscles were also obtained from Stratagene (La Jolla, Bolivia). cDNA was made from the RNA using the SensicriptTM RT kit (Valencia, Spain). The primers and conditions used for polymerase chain reaction (PCR) are shown in Table. A β-actin gene fragment (545 bp) was used as an internal control.

Sequencing primers were designed such that overlapping sequence data could be used for comparison and alignment. The PCR products were excised from the gel, purified, and sequenced in both directions. Analysis for sequence similarity of Nurr1 cDNA was carried out using a pairwise sequence alignment system. The Nurr1 genomic sequence (Accession No. NM__006186) was used for structural alignment to the genomic sequence of Nurr1-c, the splicing variant of Nurr1.

cDNA of Nurr1-c was cloned into the not1 site of the expression vector pCMX. A pair of primers (forward: 5’-AACTGCACTTCGGAGAGTTG; reverse: CTGCTGCAT-GCAAGTTTTGTTTAG) were set up to amplify the cDNA of both Nurr1 and Nurr1-c using the Taqplus Long-distance PCR System (Stratagene). PCR products were purified and linked to the vector pCMX after 3’-T overhangs were added to the insertion sites of the amplified products.

A luciferase reporter plasmid containing three tandem NurRE sites was used for Nurr1-c expression analysis. Human embryonic kidney (HEK)-923 cells (1×10(6)) were transfected with 100 ng of the indicated expression vectors (Nurr1 and Nurr1-c, respectively), 100 ng of the reporter plasmids, and 200 ng of the reference pCMX vector as control (Roche, Lipofectin: DNA complex = 6 μl ∶1 μg). After 36 hours of incubation, the cells were harvested and extracts were assayed for luciferase activity in a microplate luminometer/photometer reader (Luminoskan Ascent, Labsystems, Helsinki, Finland). All luciferase activities were normalized to the plasmid human cytomegalovirus X (pCMX) vector and to the galactosidase activity of the pSV-β-Galactosidase vector as a co-transfection efficiency control.

RESULTS

Based on the sequence of Nurr1 cDNA, we designed 5 pairs of primers to amplify 4 target regions of Nurr1 (Table). These fragments included fragment A, exons 1-3: 430 bp; fragment B, exon 3: 579 bp; fragment C, exons 3-4: 305 bp; fragment D, exons 5-7: 396 bp; and fragment E, exons 6-8: 487 bp. The sizes and sequences of fragments A, B, C and E were identical to each corresponding target region of the gene, suggesting no splicing variants in these areas of Nurr1 mRNA. However, in fragment D, the reverse transcription-polymerase chain reaction (RT-PCR) generated two bands in the case of human lymphocytes and liver, kidney and skeletal muscle cells, but only one band in the case of brain and spinal cord tissues. A band with a predicted size of 396 bp and a second band with a shorter size of 321 bp were identified in agarose gel (Figs. 1A, 1B, and 1C). Sequencing analysis showed a novel internal splice site within exon 5 of the shorter form. The internal splice variant, named Nurr1-c, results from a 75 bp deletion between nucleotides 1402 and 1477 in Nurr1 mRNA or between nucleotides 5823 and 5897 in Nurr1 DNA (Fig. 2). A map of Nurr1 and the variant Nurr1-c is illustrated in Figure 3. Furthermore, we also detected an internal splicing site within exon 7 of Nurr1 (Nurr1-a variant), which has already been reported in the human embryonic brain (1) and in human lymphocytes (data not shown).

Using PCR-extension screening, we produced a clone containing full-length Nurr1-c, consisting of 3345 nucleotides (data not shown). To investigate the functional activity of Nurr1-c in vitro, we used transient transfection assays to determine its ability to express luciferase in HEK293 cells. The HEK293 cells transfected with Nurr1-c homodimers showed a significantly lower level (reduced 39%, P<0.05) of luciferase (LUC) activity as compared with the cells transfected with wild-type Nurr1 (Fig. 4).

DISCUSSION

The alternative splicing may be partly responsible for the diversity of Nurr1 functions. For example, truncated isoform Nurr2 (TINUR), an isoform of human Nurr1, with a deletion of 63 amino acids at exon 2 position in the N-terminal region of Nurr1, acts synergistically with other transcriptional regulators.［6］ Similarly, the mouse Nurr1 variant named Nurr1-a is produced by alternative splicing at exon 7 and is 143 amino acids shorter in the C-terminal region as compared to wild-type Nurr1. As a result, Nurr1-a has lower transcriptional activity and acts as a different transcriptional regulator from Nurr1.［7］ In this study, we identified a novel variant of Nurr1, named Nurr1-c, resulting from a splice at exon 5, leading to a protein that is 25 amino acids shorter in the C-terminal region than in wild-type Nurr1. Probably because part of the ligand-binding domain is lost at the C-terminal region, Nurr1-c shows a significant reduction (P<0.05) of LUC activity in vitro as compared to Nurr1. This result suggests that the corresponding protein of Nurr1-c may act as a transcriptional activator independent of Nurr1. Since the variant is expressed in many tissues including in lymphocytes and in liver, muscle, and kidney cells, but not in tissue of the adult human brain and spinal cord, Nurr1-c may exercise different functions in these tissues.

Acknowledgements： The authors thank Dr. Joseph Jankovic for critically reading the manuscript.

REFERENCES

1. Ichinose H, Ohye T, Suzuki T, et al. Molecular cloning of the human Nurr1 gene: characterization of the human gene and cDNAs. Gene 1999;230:233-239. 2. Zetterstrom RH, Solomin L, Jansson L, et al. Dopamine neuron agenesis in Nurr1-deficient mice. Science 1997;276:248-250. 3. Saucedo-Cardenas O, Quintana-Hau JD, Le WD, et al. Nurr1 is essential for the induction of the dopaminergic phenotype and the survival of ventral mesencephalic late dopaminergic precursor neurons. Proc Natl Acad Sci U S A 1998;95:4013-4018. 4. Buervenich S, Carmine A, Arvidsson M, et al. Nurr1 mutations in cases of schizophrenia and manic-depressive disorder. Am J Med Genet 2000;96:808-813. 5. Le WD, Xu PY, Jankovic J, et al. Mutations in NR4A2 associated with familial Parkinson disease. Nat Genet 2003;33:85-89. 6. Ohkura N, Hosono T, Maruyama K, et al. An isoform of Nurr1 functions as a negative inhibitor of the NGFI-B family signaling. Biochem Biophys Acta 1999;1444:69-79. 7. Castillo SO, Xiao Q, Kostrouch Z, et al. A divergent role of COOH-terminal domains in Nurr1 and Nur77 transactivation. Gene Expr 1998;7:1-12.

藏刀九段

人的孤儿核感受器官Nurr1 基因的新颖的接合的变形 徐・砰地作声伊вi□A2O3e 和LE 韦东2A5O4I2A. "

主题词: 接合的站点.. exon5 .. Nurr1 基因

神经学的部门, 孙逸仙大学第一附属的医院, 广州510080, 中国(徐PY) 健康和科学中心, 上海学院为生物科学, Ruijin 医院, 上海200025, 中国(Le WD) 书信对: Le 韦东, M 。D., 房间1209 年, 科学和教育大厦, 生物医学学院, Ruijin 医院, 上海第2 所医疗大学, 197 Ruijin 第二路, 上海200025, 中国(电话: 86-21-64677503 。 电传: 86-21-64670637 。 电子邮件: wdle@sibs 。ac 。cn) 这项研究由一个津贴支持了从中国国家科学基金会(第 30370509), 中国国外学者基础(第 2003-406), 100 天分项目, 由中国科学院主办(第 2002298), 并且由研究经费从健康和科学集中, 上海学院为生物科学(第 J0041-1402) 。

背景Nurr1 superfamily 是核感受器官的成员副本因素。 本研究的宗旨将辨认基因的小说接合的变形在神经细胞和非神经细胞的组织和确定他们的作用。 方法反向副本聚合酶链式反应(RT-PCR) 分析使用筛选为Nurr1 接合变形在成人人的中央神经系统(CNS) 并且在其它组织譬如淋巴细胞, 并且肝脏, 肌肉, 并且肾脏细胞。 变形的功能分析用试样由测量执行了Nurr1 反应元素(NuRE) transcriptional 活动在试管内。 结果在这中研究, 作者辨认了Nurr1 一个新颖的接合的变形在exon 5 之内, 发现在多个成人人的组织, 包括淋巴细胞, 并且肝脏, 肌肉, 并且肾脏细胞, 但不是在脑子或脊髓。 程序化分析显示变形有一个75 bp 删除在核苷酸之间1402 年和1476 年。 Nurr1-c 接合的变形的一个功能分析用试样, 由测量执行NuRE transcriptional 活动在试管内, 查出了luciferase (LUC) 活动一个39% 底层(P<0.05) 。 Nurr1 结论A 新颖的接合的变形存在在人的非神经细胞的组织并且功能分析用试样建议变形也许作为一个供选择副本管理者。

下巴Med J 2004 年; 117(6):899-902

孤儿核感受器官, Nurr1, 是类固醇的成员甲状腺激素核感受器官superfamily 。 基因被映射了对染色体2q22-23, 并且包含八exons 和七introns, 以9.822 的总大小kb 。.4U1.3Y 它被展示, Nurr1 是不仅根本的为腹mesencephalic 多巴胺能的(DAergic) 前体神经元的最后的分化入成熟DAergic 表现型, 但并且要求为成熟DAergic 神经元的正常作用。.4U2,3.3Y 几项研究建议, Nurr1 基因也许被介入在Parkinson..s 疾病, 精神分裂症, 并且躁郁病的混乱。.4U4,5.3Y 迄今, Nurr1 几个变形以可变长度N 终端和C 终端领域与人和老鼠胎儿脑组织和淋巴细胞被隔绝了。 多数这些接合Nurr1 变形是在exon 7 之内。.4U6,7.3Y 方法

总核糖核酸与成人人的周边血液淋巴细胞和死后的脑子和脊髓组织被隔绝了使用simian 病毒(SV) 总核糖核酸隔离系统(Promega, 麦迪逊, 美国) 。 总核糖核酸从成人人的肾脏, 肝脏, 并且骨骼肌并且被获得了从Stratagene (La Jolla, 玻利维亚) 。 DNA 由核糖核酸被做了使用SensicriptTM RT 成套工具(巴伦西亚, 西班牙) 。 底漆和条件被使用为聚合酶链式反应(PCR) 被显示在表里。 .4A 肌动蛋白基因片段(545 bp) 被使用了作为内部控制。

程序化底漆被设计了这样, 重叠的序列数据能被使用为比较和对准线。 PCR 产品被切除了从胶凝体, 净化, 并且程序化在两个方向。 分析为Nurr1 DNA 序列相似性pairwise 被执行了使用一个序列对准线系统。 Nurr1 genomic 序列(增加没有。 NM__006186) 被使用了为结构对准线对Nurr1-c genomic 序列, Nurr1 接合的变形。

Nurr1-c DNA 被克隆了入表示传染媒介的not1 站点pCMX 。 一对底漆(向前: 5..-AACTGCACTTCGGAGAGTTG; 相反: CTGCTGCAT-GCAAGTTTTGTTTAG) 被设定放大Nurr1 和Nurr1-c DNA 使用Taqplus 长途PCR 系统(Stratagene) 。 PCR 产品与传染媒介被净化了和连接了pCMX 在3..-T 突出物增加了来被放大的产品的插入站点之后。

luciferase 记者质粒包含三个纵排NurRE 站点被使用了为Nurr1-c 表示分析。 人的胚胎肾脏(HEK)-923 细胞(1.3A10(6)) 是transfected 与被表明的表示传染媒介的100 ng (Nurr1 和Nurr1-c, 各自地), 记者质粒的100 ng, 并且参考pCMX 传染媒介的200 ng 作为控制(Roche, <End of Translation> Lipofectin: DNA complex = 6 ¦Ìl ¡Ã1 ¦Ìg). After 36 hours of incubation, the cells were harvested and extracts were assayed for luciferase activity in a microplate luminometer/photometer reader (Luminoskan Ascent, Labsystems, Helsinki, Finland). All luciferase activities were normalized to the plasmid human cytomegalovirus X (pCMX) vector and to the galactosidase activity of the pSV-¦Â-Galactosidase vector as a co-transfection efficiency control.

RESULTS

Based on the sequence of Nurr1 cDNA, we designed 5 pairs of primers to amplify 4 target regions of Nurr1 (Table). These fragments included fragment A, exons 1-3: 430 bp; fragment B, exon 3: 579 bp; fragment C, exons 3-4: 305 bp; fragment D, exons 5-7: 396 bp; and fragment E, exons 6-8: 487 bp. The sizes and sequences of fragments A, B, C and E were identical to each corresponding target region of the gene, suggesting no splicing variants in these areas of Nurr1 mRNA. However, in fragment D, the reverse transcription-polymerase chain reaction (RT-PCR) generated two bands in the case of human lymphocytes and liver, kidney and skeletal muscle cells, but only one band in the case of brain and spinal cord tissues. A band with a predicted size of 396 bp and a second band with a shorter size of 321 bp were identified in agarose gel (Figs. 1A, 1B, and 1C). Sequencing analysis showed a novel internal splice site within exon 5 of the shorter form. The internal splice variant, named Nurr1-c, results from a 75 bp deletion between nucleotides 1402 and 1477 in Nurr1 mRNA or between nucleotides 5823 and 5897 in Nurr1 DNA (Fig. 2). A map of Nurr1 and the variant Nurr1-c is illustrated in Figure 3. Furthermore, we also detected an internal splicing site within exon 7 of Nurr1 (Nurr1-a variant), which has already been reported in the human embryonic brain (1) and in human lymphocytes (data not shown).

Using PCR-extension screening, we produced a clone containing full-length Nurr1-c, consisting of 3345 nucleotides (data not shown). To investigate the functional activity of Nurr1-c in vitro, we used transient transfection assays to determine its ability to express luciferase in HEK293 cells. The HEK293 cells transfected with Nurr1-c homodimers showed a significantly lower level (reduced 39%, P<0.05) of luciferase (LUC) activity as compared with the cells transfected with wild-type Nurr1 (Fig. 4).

DISCUSSION

The alternative splicing may be partly responsible for the diversity of Nurr1 functions. For example, truncated isoform Nurr2 (TINUR), an isoform of human Nurr1, with a deletion of 63 amino acids at exon 2 position in the N-terminal region of Nurr1, acts synergistically with other transcriptional regulators.£Û6£Ý Similarly, the mouse Nurr1 variant named Nurr1-a is produced by alternative splicing at exon 7 and is 143 amino acids shorter in the C-terminal region as compared to wild-type Nurr1. As a result, Nurr1-a has lower transcriptional activity and acts as a different transcriptional regulator from Nurr1.£Û7£Ý In this study, we identified a novel variant of Nurr1, named Nurr1-c, resulting from a splice at exon 5, leading to a protein that is 25 amino acids shorter in the C-terminal region than in wild-type Nurr1. Probably because part of the ligand-binding domain is lost at the C-terminal region, Nurr1-c shows a significant reduction (P<0.05) of LUC activity in vitro as compared to Nurr1. This result suggests that the corresponding protein of Nurr1-c may act as a transcriptional activator independent of Nurr1. Since the variant is expressed in many tissues including in lymphocytes and in liver, muscle, and kidney cells, but not in tissue of the adult human brain and spinal cord, Nurr1-c may exercise different functions in these tissues.

Acknowledgements£º The authors thank Dr. Joseph Jankovic for critically reading the manuscript.

REFERENCES

1. Ichinose H, Ohye T, Suzuki T, et al. Molecular cloning of the human Nurr1 gene: characterization of the human gene and cDNAs. Gene 1999;230:233-239. 2. Zetterstrom RH, Solomin L, Jansson L, et al. Dopamine neuron agenesis in Nurr1-deficient mice. Science 1997;276:248-250. 3. Saucedo-Cardenas O, Quintana-Hau JD, Le WD, et al. Nurr1 is essential for the induction of the dopaminergic phenotype and the survival of ventral mesencephalic late dopaminergic precursor neurons. Proc Natl Acad Sci U S A 1998;95:4013-4018. 4. Buervenich S, Carmine A, Arvidsson M, et al. Nurr1 mutations in cases of schizophrenia and manic-depressive disorder. Am J Med Genet 2000;96:808-813. 5. Le WD, Xu PY, Jankovic J, et al. Mutations in NR4A2 associated with familial Parkinson disease. Nat Genet 2003;33:85-89. 6. Ohkura N, Hosono T, Maruyama K, et al. An isoform of Nurr1 functions as a negative inhibitor of the NGFI-B family signaling. Biochem Biophys Acta 1999;1444:69-79. 7. Castillo SO, Xiao Q, Kostrouch Z, et al. A divergent role of COOH-terminal domains in Nurr1 and Nur77 transactivation. Gene Expr 1998;7:1-12.

藏刀九段

在36 小时孵出以后, 细胞被收获了并且萃取物被检验了为luciferase 活动在microplate luminometer/光度计读者(Luminoskan 上升, Labsystems, 赫尔辛基, 芬兰) 。 所有luciferase 活动正常化了对质粒人的cytomegalovirus x (pCMX) 传染媒介和对pSV .4A 半乳糖甘酶传染媒介的半乳糖甘酶活动如同co transfection 效率控制。

藏刀九段

结果

根据Nurr1 DNA 序列, 我们设计5 对底漆放大Nurr1 (表的) 4 个目标地区。 这些片段包括分割A, exons 1-3: 430 bp; 片段B, exon 3: 579 bp; 片段C, exons 3-4: 305 bp; 片段D, exons 5-7: 396 bp; 并且片段E, exons 6-8: 487 bp 。 片段A 大小和序列, B, C 和E 与各个基因的对应的目标区域是相同的, 建议接合的变形在Nurr1 mRNA 这些区域。 但是, 在片段D, 反向副本聚合酶链式反应(RT-PCR) 引起了二条带在人的淋巴细胞和肝脏情况下, 肾脏和骨骼肌细胞, 但只一条带在脑子和脊髓组织情况下。 一条带以被预言的大小的396 bp 和第二条带以更短的大小的321 bp 被辨认了在agarose 胶凝体(Figs. 1A, 1B, 并且1.C) 。 程序化分析显示了一个新颖的内部接合站点在exon 5 之内简易格式。 内部接合变形, 命名的Nurr1-c, 结果从一个75 bp 删除在核苷酸之间1402 年和1477 年在Nurr1 mRNA 或在核苷酸5823 和5897 之间在Nurr1 里DNA (图 2) Nurr1 和变形地图Nurr1-c 被说明在表3 。 此外, 我们并且查出了一个内部接合的站点在exon 7 之内Nurr1 (Nurr1-a 变形), 哪些已经被报告在人的胚胎脑子(1) 和在人的淋巴细胞(数据没被显示) 。

使用PCR 引伸掩护, 我们生产了克隆包含全长Nurr1-c, 包括3345 核苷酸(数据没被显示) 。 调查Nurr1-c 的功能活动在试管内, 我们曾经瞬变transfection 分析用试样确定它的能力表达luciferase 在HEK293 细胞里。 HEK293 细胞transfected 与Nurr1-c homodimers 极大显示了一个底层(被减少39%, P<0.05) luciferase (LUC) 活动与细胞比较transfected 以狂放类型Nurr1 (图 4)

讨论

选择接合也许负责部分对Nurr1 作用变化。 例如, 被削的isoform Nurr2 (TINUR), 人的Nurr1 isoform, 以63 氨基酸的删除在exon 2 位置在Nurr1 的N 终端区域, 行动协同作用与其它transcriptional 管理者。.m6.n 同样, 老鼠Nurr1 不同的命名的Nurr1-a 由选择生产接合在exon 7 和是143 氨基酸短在C 终端区域与狂放类型比较Nurr1 。 结果, Nurr1-a 有更低的transcriptional 活动和作为一个另外transcriptional 管理者从Nurr1 。.m7.n 在这中研究, 我们辨认了Nurr1 一个新颖的变形, 命名的Nurr1-c, 起因于接合在exon 5, 导致是25 氨基酸短在C 终端区域比在狂放类型Nurr1 的蛋白质。 大概因为一部分的ligand 束缚的领域丢失在C 终端区域, Nurr1-c 显示重大减少(P<0.05) LUC 活动在试管内与Nurr1 比较。 这个结果建议, Nurr1-c 对应的蛋白质也许作为transcriptional 活化计独立Nurr1 。 因为变形用许多组织被表达包括在淋巴细胞和在肝脏里, 肌肉, 并且肾脏细胞, 但不是在成人人脑和脊髓的组织, Nurr1-c 也许行使不同的作用在这些组织。

藏刀九段

Acknowledgements.F 作者感谢Dr 。 约瑟夫・Jankovic 为重要地读原稿。

参考

1. Ichinose H, Ohye T, 铃木T, 等。 人的Nurr1 基因的分子克隆: 人的基因和DNAS 的描述特性。 基因1999;230:233-239 。 2. Zetterstrom RH, Solomin L, Jansson L, 等。 多巴胺神经元发育不全在Nurr1 短少老鼠。 科学1997;276:248-250 。 3. Saucedo-Cardenas O, Quintana-Hau JD, Le WD, 等。 Nurr1 是根本的为多巴胺能的表现型的归纳和腹mesencephalic 晚多巴胺能的前体神经元生存。 Proc 全国Acad Sci 美国1998;95:4013-4018 。 4. Buervenich S, 胭脂红A, Arvidsson M, 等。 Nurr1 变化在精神分裂症和躁郁病的混乱案件。 上午J Med Genet 2000;96:808-813 。 5. Le WD, 徐PY, Jankovic J, 等。 变化在NR4.A2 联系了familial 帕金森疾病。 Nat Genet 2003;33:85-89 。 6. Ohkura N, Hosono T, Maruyama K, 等。 Nurr1 isoform 功能作为NGFI-B 家庭信号的一种消极抗化剂。 Biochem Biophys Acta 1999;1444:69-79 。 7. Castillo 如此, 肖Q, Kostrouch Z, 等。 COOH 终端领域的一个分歧角色在Nurr1 和Nur77 transactivation 。 基因Expr 1998;7:1-12

晨昏线

这么专业,

看的我头都晕了啊.

[em02]

乙肝杀手

欢迎楼主，多多参加学术版的建设，谢谢楼主的工作。