人脑低级别星形细胞瘤及瘤周脑组织的差异蛋白表达

Differential protein expression between various pathological grades of glioma has been shown in studies of glioma proteomics. However, very little data is available regarding normal brain tissues and glioma differential protein expression, because normal human brain tissues are difficult to harvest. The present study selected samples from low-grade astrocytomas and peritumoral brain tissues to analyze differential protein expression by two-dimensional (2D) electrophoresis and mass spectrometry techniques. Results revealing 36 protein spots by 2D electrophoresis, including 23 spots revealing increased expression and 13 spots revealing decreased expression. However, 25 differential proteins were identified by mass spectrometry, including 16 proteins with increased expression and 9 with decreased expression. Western blot analysis confirmed the mass spectrometry results, i.e., heat shock protein 70 (HSP70) and human transthyretin (TTR) expressions were increased, but glial fibrillary acidic protein (GFAP) was decreased, in astrocytomas. The present study constructed a 2D electrophoresis pattern between low-grade astrocytomas in the human brain and peritumoral tissues. Results demonstrated that a majority of differential proteins, such as HSP70, TTR, and GFAP, participate in malignant progression of gliomas.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

Proteomic identification of brainstem cytosolic proteins in a neuropathic pain model

Neuropathic pain involves co-regulation of many genes and their translational products in both peripheral and central nervous system. We used proteomics approaches to investigate expressional changes in cytosolic protein levels in rat brainstem tissues following ligation of lumbar 5 and 6 (L5, L6) spinal nerves, which generates a model of peripheral neuropathic pain (NP). Proteins from brainstem tissue homogenates of NP and SHAM animals were fractionated by two-dimensional (2-DE) gel electrophoresis to produce a high-resolution map of the brainstem soluble proteins. Proteins showing altered expression levels between NP and SHAM were selected. Isolated proteins were in-gel trypsin-digested and the resulting peptides were analyzed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. Using the mass spectrometric data, we were able to identify 17 proteins of interest through searches of the Swiss-Prot and NCBi nonredundant protein sequence database. Several of the identified proteins, including fatty acid binding protein-brain (FABP-B), major histocompatibility complex (MHC) class 1, T-cell receptor (TCR) alpha chain, and interleukin-1 (IL-1), showed significantly higher levels in the NP rat brainstem. Proteomic analysis has identified several proteins with differential expression levels in NP as compared to SHAM. However, the function of the proteins identified is postulated; therefore, further experiments are required to determine the true role of each protein in NP.     

原发性脑创伤后人脑皮层差异蛋白质组研究

目的应用蛋白质组学技术，研究不同程度脑创伤后的人脑皮层蛋白质组表达变化的情况。方法将临床标本以GCS评分分为中度和重度损伤组，提取脑挫伤部位皮层的总蛋白。通过双向电泳．分离蛋白。应用胶内酶切、生物质谱，鉴定由图像分析软件所得出的具有表达差异的蛋白质点。结果通过比较，目前已发现12个蛋白质点，表达水平具有显著性差异变化。在已鉴定出的蛋白点中，属于10种蛋白质。依其功能可分为：细胞骨架、代谢反应、氧化应激反应、功能未知等几类。在重伤组中，一共有9个蛋白点、6种蛋白表达上调（α-烯醇化酶、磷酸丙糖异构酶、5’磷酸吡哆醇氧化酶、crtstalin、stathmin-1、NP25）；中度损伤组3个蛋白点、4种蛋白表达上调（谷氨酰胺S转移酶、5’3’核苷酸酶、HSPC108、proapolipoprotein）。结论外伤后，神经系统由于受伤程度的不同，蛋白表达水平会发生变化。原发性脑创伤的损伤程度不同将产生神经系统病生理反应的差异。     

抑郁模型大鼠小脑的比较蛋白组学初步研究

目的 通过抑郁模型大鼠小脑的差异蛋白组学分析,探讨小脑在抑郁症病理生理过程中的作用.方法 建立慢性不可预见性温和应激(CUMS)抑郁大鼠模型,并应用同位素标记相对和绝对定量技术结合多维液相色谱-串联质谱分析CUMS组和对照组大鼠小脑差异蛋白质.随后对结果采用DAVID生物信息数据库进行分析.结果 与对照组大鼠比较,CUMS组大鼠糖水偏好值明显降低,差异有统计学意义(P＜0.05).对两组样品鉴定,共得到差异蛋白55个,其中在CUMS组中表达下调的有18个,表达上调的有37个.这些蛋白主要参与神经递质代谢、糖酵解、ATP合成利用、脂质代谢、氨基酸代谢等生物过程.结论 本研究所发现的差异蛋白可能是抑郁症小脑功能异常的神经生物学基础,对深入探讨小脑在抑郁症发病机制中的作用提供了重要的线索.     

Impact of thyroid hormone deficiency on the developing CNS: cerebellar glial and neuronal protein expression in rat neonates exposed to antithyroid drug propylthiouracil

The developing rat cerebellum is vulnerable to thyroid hormone (TH) deficiency. The present study addresses the molecular mechanisms involved in this response. Specifically, the study focuses on the expression of selected cerebellar proteins that are known to be directly [protein expressing 3-fucosyl-N-acetyl-lactosamine antigen (CD15), neuronal cell adhesion molecule (L1)] or indirectly [glial fibrillary acidic protein (GFAP)], involved in glial-neuronal interactions and thus regulation of cell proliferation and granule cell migration. Cerebellar mass, structure, and protein expression in rat neonates exposed to antithyroid drug propylthiouracil (PTU) from the embryonic day (E) 16 to postnatal day (P) 21 were compared against rat neonates that received replacement of thyroxin (T4) starting on day P1 or untreated controls. Cerebellar proteins were analyzed by quantitative Western blots. PTU-treated rats lagged in growth and showed reduction in cerebellar mass and alterations in cerebellar structure on P15. Daily treatment of neonates with T4 restored normal cerebellum-to-body-mass ratio, cerebellar structure, and cerebellar protein expression. Densitometric analysis of Western blots revealed altered expression of selected proteins in the cerebella of hypothyroid neonates. A decrease of CD15 (46%, p = 0.031) was observed on P10 and was accompanied by a decrease in GFAP expression (64%, p = 0.039). Furthermore, a shift in the developmental GFAP profile was observed in the PTU-treated cerebellum. L1 expression was not significantly affected in the hypothyroid cerebellum. Altered expression of cerebellar proteins is likely to affect cell-cell interactions and consequently cell proliferation and migration and contribute to structural and functional alterations seen in the hypothyroid rat neonates.     

cDNA cloning and characterization of three genes uniquely expressed in cerebellum by Purkinje neurons

The characteristics that distinguish the different neuronal cell types of an organism are believed to be primarily determined by unique patterns of cellular gene expression. The identification of cell-type specific molecules should therefore provide a good basis for understanding the biology of specific neuron types. In this paper, we describe the isolation of cDNA clones corresponding to mRNA uniquely expressed by Purkinje cells in mature mouse cerebellum. Three cDNA clones were selected from a library of normal mouse cerebellar cDNA by virtue of their failure to hybridize to mRNA sequences from the cerebella of Purkinje cell degeneration (pcd) mice. The cDNA clones were shown by in situ and Northern hybridization to correspond with mRNA present in Purkinje cells but absent or at low levels in other cell types of the cerebellum. By sequence analysis, clone PCD29 was determined to encode the calcium-binding protein calbindin-D28K. Clones PCD5 and PCD6 encode previously undescribed proteins of 99 and greater than 500 amino acids, respectively. All 3 PCD clones hybridized to mouse mRNA from sources other than cerebellum; clone PCD5 was found to have the most restricted expression, as it hybridized only to mRNA from cerebellum and eye. To define potential correlations between the PCD clones and mutations in the mouse genome known to affect Purkinje cells, clones PCD5, PCD6, and PCD29 were localized to mouse chromosomes 8, 6, and 4, respectively.     

Distribution of Purkinje cell-specific Zebrin-II/aldolase C immunoreactivity in the mouse,rat, rabbit,and human retina

The developmental, genetic, and biochemical similarities that have been observed between the cerebellum and retina form the basis for ongoing investigations into retinal expression of cerebellar-specific proteins. We have examined the mouse, rat, rabbit, and human retina for expression of a protein that is present in parasagittal Purkinje cell strips and that is recognized by the antibody Zebrin-II. This protein has recently been identified as a member of the aldolase C isoenzymes. Western blotting and immunocytochemistry have been used. The monoclonal antibody Zebrin-II recognized a prominent 36 kDa protein band on immunoblots of both the cerebellum and the retina of the examined species. Immunocytochemistry showed that, in the three nonhuman species, cells were stained in the ganglion cell layer (GCL). In addition, in the mouse and rabbit, cells in the inner nuclear layer (INL) were also labeled. Except for the visual streak, there were more immunopositive cells in the rabbit GCL and INL than in corresponding areas of the mouse retina. In the human, in contrast to the other species, the photoreceptor cell layer was strongly aldolase C immunoreactive. In the h all species except for the rat, the photoreceptor inner segments also displayed a weak labeling. The results show that this aldolase C isoenzyme is another protein that is selectively expressed by the cerebellum: and retina. Furthermore, the retinal expression is species specific, and this pattern seems to show a good correlation with the oxygenation level of the individual compartments. The indication that this aldolase C isoenzyme has specific developmental functions in the retina provides additional clues for our understanding of cerebellar organization. © 1994 Wiley-Liss, Inc.     

Selective reduction in synaptic proteins involved in vesicle docking and signalling at synapses in the ataxic mutant mouse stargazer

The spontaneous recessive mutant mouse stargazer has a specific and pronounced deficit in brain-derived neurotrophic factor (BDNF) mRNA expression in the cerebellum. Cerebellar granule cells, in particular, show a selective and near-total loss of BDNF. The mutation involves a defect in the calcium channel subunit Cacng2. This severely reduces expression of stargazin. A stargazin-induced failure in BDNF expression is thought to underlie the cerebellar ataxia with which the mutant presents. BDNF is known to regulate plasticity at cerebellar synapses. However, relatively little is known about the mechanism involved. We previously demonstrated that the stargazer mutation affects the phenotype of cerebellar glutamatergic neurons. Stargazer neurons have less glutamate and proportionally fewer docked vesicles at presynaptic sites than controls. In the current study, we investigate the mechanism underlying BDNF-induced synaptic changes by analyzing alterations in synaptic signalling proteins in the stargazer cerebellum. Expression levels of synaptic proteins were evaluated by measuring relative density of immunogold label over granule cell terminals in ultrathin sections from ataxic stargazer mutants compared with matched nonataxic littermates. We show that there is a selective and marked depletion in the levels of vesicle-associated proteins (synaptobrevin, synaptophysin, synaptotagmin, and Rab3a) but not of plasma membrane-associated protein (SNAP-25) in the terminals of the BDNF-deficient granule cells. Changes are restricted to the cerebellum; levels in the hippocampus are unaltered. These data suggest that the BDNF deficits in the cerebellum of stargazer affect synaptic vesicle docking by selectively altering synaptic-protein distribution and abundance.