GENE EXPRESSION ANALYSIS OF THE SPINAL CORD UNDER CHRONIC MECHANICAL COMPRESSION USING IN-HOUSE COMPLEMENTARY DNA MICROARRAY

使用内部互补 DNA 微阵列对慢性机械压迫下的脊髓进行基因表达分析

• 批准号: 15591563
• 负责人: OKAWA Akihiko
• 金额: $ 1.34万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591563/
• 关键词: chronic spinal cord compression; cDNA microarray; RT-PCR; ttw mice; cDNAマイクロアレイ; 慢性圧迫性脊髄障害

To elucidate precise mechanism of progression of chronic compressive myelopathy, we analyzed gene expression profile of chronically-compressed spinal cord using cDNA microarray. We employed ttw mice, which is a mutant mouse naturally develop calcified deposit at C1/2 level, as a model for chronic spinal cord compression. Fluorescent probe were made from mRNA extracted from spinal cord tissue of ttw or control mice and hybridized with cDNA microarray which contains cDNA from mouse brain and neural stem cell. After the identification of differentially expressed genes in the spinal cords of the ttw and control mice by our microarray system, we confirmed the results using semi-quantitative RT-PCR. Among the 4,851 genes on the cDNA microarray chip, one gene (0.02%) showed increase greater than a 2-fold, and 17 genes (0.35%) showed decrease less than a 2-fold in expression. The up-regulated gene was gephyrin (X66366). The seventeen down-regulated genes were extracellular superoxide dismutase … More (SOD3;U38261), p34 (AF178669), GalNAc alpha-2,6-sialyltransferase V (AB028840), a glucosidase II a subunit (U92793), nuclear RNA export factor 1 homolog (BC005594), ss-spectrin (Spnb-2;M74773), UDP-N-acetylglucosaminyltransferase (AF363030), mPACPL1 (AB030038), HOOK1 (AF044923), NCAML1 (AU035962), NSPC1 (BC004952), solute carrier family 29 (BC006812), serine/threonine protein kinase (AB041542), and 4 unknown genes (BE291425,BC003481,AU051226,AU078872). To confirm the microarray data, semi-quantitative RT-PCR was performed for the microarray-screened 18 genes. Among those 18 genes, gephyrin (up-regulated in ttw mice) and NSPC1 (down-regulated in ttw mice) were excluded because no significant difference in expression level of those two genes was detected by semi-quantitative RT-PCR. On the contrary to the results of our previous study about acute spinal cord injury, unexpectedly small number of genes showed alteration of expression in the chronic spinal cord compression in the present study. There may be difference in mechanism of progression between acute and chronic spinal cord compression. Less

为了阐明慢性压迫性脊髓病进展的确切机制，我们使用cDNA微阵列分析了慢性压迫性脊髓的基因表达谱。我们采用ttw小鼠作为慢性脊髓压迫模型，ttw小鼠是一种在C1/2水平自然形成钙化存款的突变小鼠。从ttw小鼠和对照小鼠的脊髓组织中提取mRNA，制成荧光探针，与含有小鼠脑和神经干细胞cDNA的cDNA微阵列杂交。在我们的微阵列系统鉴定ttw和对照小鼠脊髓中差异表达的基因后，我们使用半定量RT-PCR证实了结果。在cDNA微阵列芯片上的4，851个基因中，1个基因（0.02%）显示表达增加大于2倍，17个基因（0.35%）显示表达减少小于2倍。上调基因为桥蛋白（X66366）。下调的17个基因为胞外超氧化物歧化酶 ...更多信息 (SOD3;U38261），第34页（AF 178669），GalNAc α-2，6-唾液酸转移酶V（AB 028840），葡萄糖苷酶II a亚基（U92793），核RNA输出因子1同源物（BC 005594），ss-血影蛋白（Spnb-2; M74773），UDP-N-乙酰葡糖胺转移酶（AF363030），mPACPL1（AB 030038），钩1（AF044923），NCAML1（AU035962），NSPC1（BC 004952）、溶质载体家族29（BC 006812）、丝氨酸/苏氨酸蛋白激酶（AB 041542）和4个未知基因（BE 291425、BC 003481、AU 051226、AU 078872）。为了证实微阵列数据，对微阵列筛选的18个基因进行半定量RT-PCR。在这18个基因中，排除了gephyrin（在ttw小鼠中上调）和NSPC 1（在ttw小鼠中下调），因为通过半定量RT-PCR检测到这两个基因的表达水平没有显著差异。与我们以前关于急性脊髓损伤的研究结果相反，在本研究中，出乎意料的少数基因在慢性脊髓压迫中表现出表达改变。急性和慢性脊髓压迫症的进展机制可能存在差异。少

项目成果

Anomalous vertebral artery at the craniovertebral junction in a patient with Down Syndrome : A case report.

唐氏综合症患者颅椎交界处椎动脉异常：病例报告。

• 发表时间: 2004
• 期刊: Journal of Neurosurgery Spine 1号3巻
• 作者: Ymazaki M;Koda M;et al.
• 通讯作者: et al.

Gene expression profiling of cathepsin D, metallothioneins-1 and-2, osteopontin, and tenascin-C in a mouse spinal cord injury model by cDNA microarray analysis

• DOI: 10.1007/s00401-004-0926-z
• 发表时间: 2005-02-01
• 期刊: ACTA NEUROPATHOLOGICA
• 影响因子: 12.7
• 作者: Hashimoto, M;Koda, M;Moriya, H
• 通讯作者: Moriya, H

Hashimoto M, Yamazaki M 他: "Upregulation of osteopontin expression in rat spinal cord microglia after traumatic injury"Journal of Neurotrauma. 20(3). 287-296 (2003)

Hashimoto M、Yamazaki M 等人：“创伤性损伤后大鼠脊髓小胶质细胞中骨桥蛋白表达的上调”《神经创伤杂志》20(3) (2003)。

Transplanted hematopoietic stem cells form bone marrow differentiate into neural lineage cells and promote functional recovery after spinal cord injury in mice.

移植的造血干细胞在小鼠脊髓损伤后形成骨髓分化为神经谱系细胞并促进功能恢复。

• 发表时间: 2004
• 期刊: J Neuropathol Exp Neurol 63巻
• 作者: Koshizuka S;Okawa A;et al.
• 通讯作者: et al.

Yonghui Fu, Yainazaki M 他: "Spinal root avulsion-induced upregulation of osteopontin expression in the adult rat spinal cord"Acta Neuropathologica. 107. 8-16 (2004)

Yonghui Fu, Yainazaki M et al.:“成年大鼠脊髓中脊髓根撕脱诱导的骨桥蛋白表达上调”Acta Neuropathologica. 107. 8-16 (2004)