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Identification and Characterization of a Rat Novel Gene RSEP4 Expressed Specifically in Central Nervous System

Acta Biochimica et Biophysica Sinica 2004年第7期36卷 501-507页

作者： Xi-Dao WANG1,2, Ling-Wei KONG1, Zhi-Qin XIE1, Yu-Qiu ZHANG3, Zhi-Xin LIN2, Zhi-Qi ZHAO3,Lei YU4, and Nai-He JING1* 1 Laboratory of Molecular cell biology, Laboratory of Stem cell biology, Institute of Biochemistry and cell biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China School of Life Science and Biotechnology, 2 Shanghai Jiaotong university, Shanghai 200030, China Institute of neurobiology, Fudan university, Shanghai 200433, China 3 4 department of cell biology, neurobiology and anatomy, college of medicine, university of cincinnati, cincinnati, ohio, usa Laboratory of Molecular cell biology and Stem cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences Shanghai China

The low-abundantly expressed genes composed the majorities of the mRNAs expressed in the central nervous system (CNS), and were thought to be important for the normal brain functions. Through differential screening a low-abundance cDNA sublibrary with mRNA from neuropathic pain of chronic constriction injury (CCI) model, we have identified a novel rat gene, rat spinal-cord expression protein 4 gene (RSEP4). The total length of RSEP4 cDNA is 2006 bp, with a 501 nucleotide open reading frame (ORF) that encodes a 167 amino acid polypeptide. Northern blot revealed that RSEP4 was expressed specifically in the CNS. In situ hybridization showed that the mRNA of RSEP4 was strongly expressed in the CA1, CA2, CA3 and DG regions of hippocampus, the Purkinje cells of cerebellum, and the small sensory neurons of dorsal horn and large motor neurons of ventral horn of spinal cord. Over-expression of RSEP4-EGFP fusion protein in the human embryonic kidney 293T cells showed that RSEP4 protein was mainly localized in the cell cytoplasm. These results suggest that RSEP4 may play some roles in the CNS.

关键词： central nervous system (CNS) rat Northern blot in situ hybridization

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DNA fragmentation in apoptosis

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cell Research 2000年第3期10卷 205-211页

作者： ZHANG JIAN HUA, MING XU (department of cell biology, neurobiology and anatomy, university of cincinnati college of medicine, cincinnati, ohio 452670521, usa) departmentofcellbiology NeurobiologyandAnatomyUniversityofCincinnatiCollegeofMedicineOhio45267-0521USA.

Cleavage of chromosomal DNA into oligonucleosomal size fragments is an integral part of apoptosis. Elegant biochemical work identified the DNA fragmentation factor (DFF) as a major apoptotic endonuclease for DNA fragmentation in vitro. Genetic studies in mice support the importance of DFF in DNA fragmentation and possibly in apoptosis in vivo. Recent work also suggests the existence of additional endonucleases for DNA degradation. Understanding the roles of individual endonucleases in apoptosis, and how they might coordinate to degrade DNA in different tissues during normal development and homeostasis, as well as in various diseased states, will be a major research focus in the near future.

关键词： DNA fragmentation DFF endonucleases apop tosts.

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Mitochondrial DNA Mutations Associated with Aminoglycoside Ototoxicity

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Journal of Otology 2006年第2期1卷 65-75页

作者： GUAN Min-Xin Division of Human Genetics and Center for Hearing and Deafness Research, cincinnati Children’s Hospital Medical Center, cincinnati, ohio 45229, usa department of Pediatrics, university of cincinnati college of medicine, cincinnati, ohio 45229, usa Division of Human Genetics and Center for Hearing and Deafness Research Cincinnati Children’s Hospital Medical Center Cincinnati Ohio 45229 USA department of Pediatrics University of Cincinnati College of Medicine Cincinnati Ohio 45229 USA

The mitochondrial 12S rRNA has been shown to be the hot spot for mutations associated with both aminoglycoside-induced and non-syndromic hearing loss. Of all the mutations, the homoplasmic A1555G and C1494T mutations at a highly conserved decoding region in the 12S rRNA have been associated with aminoglycoside-induced and non-syndromic hearing loss in many families worldwide. The A1555G or C1494T mutation is expected to form novel 1494C-G1555 or 1494U-A1555 base-pair at the highly conserved A-site of 12S rRNA. These transitions make the secondary structure of this RNA more closely resemble the corresponding region of bacterial 16S rRNA. Thus, the new U-A or G-C pair in 12S rRNA created by the C1494T or A1555G transition facilitates the binding of aminoglycosides, thereby accounting for the fact that the exposure to aminoglycosides can induce or worsen hearing loss in individuals carrying these mutations. Furthermore, the growth defect and impairment of mitochondrial translation were observed in cell lines carrying the A1555G or C1494T mutation in the presence of high concentration of aminoglycosides. In addition, nuclear modifier genes and mitochondrial haplotypes modulate the phenotypic manifestation of the A1555G and C1494T mutations. These observations provide the direct genetic and biochemical evidences that the A1555G or C1494T mutation is a pathogenic mtDNA mutation associated with aminoglycoside-induced and nonsyndromic hearing loss. Therefore, these data have been providing valuable information and technology to predict which individuals are at risk for ototoxicity, to improve the safety of aminoglycoside antibiotic therapy, and eventually to decrease the incidence of deafness.

关键词： gene Mitochondrial DNA Mutations Associated with Aminoglycoside Ototoxicity rRNA DNA

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An increase in opening of BKCa channels in smooth muscle cells in streptozotocin-induced diabetic mice

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Acta Pharmacologica Sinica 2004年第6期25卷 42-48页

作者： Chun-ling YE2, Bing SHEN4, Xian-da REN, Rong-jing LUO3, Sheng-yuan DING3, Fu-man YAN3, Jia-hua JIANG department of Pharmacology, Pharmacy college of Ji-nan university, Guangzhou 510632 3department of Physiology, Guangzhou university of Traditional Chinese medicine, Guangzhou 510407 4department of Molecular biology and cell biology, School ofLife Sciences, university of Sciences and Technology of China, Hefei 230036, China department of Pharmacology Pharmacy College of Ji-nan University Guangzhou 510632 Chin

AIM: To investigate the changes of function of large conductance of calcium-activated potassium channels (BKCa channels) in thoracic aortic smooth muscle cells in early stage of streptozotocin (STZ)-induced diabetic C57BL/6J mice. METHODS: Vascular muscle tension in the isolated thoracic aortic rings of mice was compared, and the role of BKCa channels in relaxation of isolated mice thoracic aortic rings induced by acetylcholine (ACh) was determined. Meanwhile, single vascular smooth muscle cells (VSMCs) were isolated by collagenase, and BKCa currents were recorded by patch-clamp single channel recording technique in symmetric high potassium solution. RESULTS: Tetraethylammonium (TEA) 1 mmol/L, a selective calcium-activated potassium channel blocker, caused significant rightward shift in the concentration-response curves of ACh in the isolated thoracic aortic rings of diabetic mice and pD2 value of ACh-induced relaxation was decreased notably after TEA treatment [(6.3±0.4) vs (6.9±0.5), n=10 rings from 7 mice, P4±0.15) vs (6.5±0.5), n=7 rings from 6 mice, P>0.05]. Furthermore, conductance of BKCa channels in single thoracic aortic smooth muscle cells was decreased [(199±15) pS, n=10 cells from 7 mice vs (266±11) pS, n=12 cells from 6 mice, Pcells from 6 mice, P32±98), n=6 cells from 6 mice, Pcells in the early stage of STZ-induced diabetic C57BL/6J mice by reducing mean closed time, but the conductance of BKCa channels was decreased.

关键词： diabetes mellitus streptozotocin aorta patch-clamp techniques potassium channels calcium

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Manipulation of collagen synthesis influences progenitor cell engraftment and angiomyogenesis

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岭南心血管病杂志 2011年第S1期17卷 32-33页

作者： Yigang Wang (department of Pathology and Laboratory medicine, college of medicine,university of cincinnati Medical Center,cincinnati,ohio 45267,usa)

Myocardial infarction(MI)results in loss of cardiomyocytes(CM) in the ischemic area of the heart followed by an inflammatory response and replacement of contractile CM with *** fibrosis,a key contributor to cardiac dysfunction after MI,presents as a secondary response to the pathophysiological remodeling of long-standing disease including ischemia,obstruction,and microvascular *** fibroblasts and myofibroblasts are responsible for post-MI remodeling which occurs via regulation of extracellular matrix (ECM),presenting as increased collagenⅠandⅢinto the interstitial and perivascular *** addition to the pluripotency of stem cells following stem/ progenitor cell transplantation,decreased apoptosis, hypertrophy,and fibrosis in the infarcted heart have been *** has made transplantation of progenitor/stem cells a primary research focus in the field of tissue ***,the accumulation of ECM and myofibroblasts in areas of tissue injury presents a barrier that can impair penetration of reparative stem/progenitor cells mobilized from peripheral ***,cardiac fibroblast production and degradation of ECM are critical in regulating cardiac remodeling and stem/progenitor cell *** study used transgenic mice overexpressing adenylyl cyclaseⅥ(AC6) in which collagen synthesis was decreased to determine the role of collagen deposition on the engraftment of iPSC from a tri-cell patch applied to infarcted area after MI.

关键词： cell Manipulation of collagen synthesis influences progenitor cell engraftment and angiomyogenesis iPSC MEF ECM

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Signaling, Actin Dynamics and Endocytosis

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生物物理学报 2001年第1期17卷 10-18页

作者： SONG Wen-xia(department of cell biology and Molecular Genetics, university of Maryland, college Park, MD 20742, usa) university of Maryland 美国 20742

Endocytosis is an essential process for normal function of all living cells. cells get nutrients, and control the surface-expressional level of proteins as well as membrane hemostats through the endocytosis. Endocytosis process is regulated in response to functional status of a particular cell. Signaling events and the endocytosis process go hand in hand to fulfill cellular functions. Although our understanding of the endocytosis process has grown rapidly during the last decade, little is known about how it is interconnected functionally with the signaling status of cells. During endocytosis, vesicles are formed from the plasma membrane through complex molecular machinery. The location where the vesicles are formed is rich in cortical actin cytoskeleton that supports the plasma membrane. To enter cells, vesicles have to diffuse through the cortical actin cytoskeleton. The actin cytoskeleton has a very dynamic structure and actively participates a wide variety of cellular functions. In addition to its central role in cytokinesis, cell shape, cell motility, and cell polarity, a connection between the endocytosis process and the actin cytoskeleton has been implicated in both yeast and mammalian system. In recent years the knowledge on how the actin cytoskeleton participates in the generation of coordinated cellular responses to external stimuli is grown rapidly. In this review, we focus on the potential roles of the actin cytoskeleton in regulating the endocytosis process in response to signaling events.

关键词：信号肌动蛋白胞吞

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Prevention of pathological change and cognitive degeneration of Tg2576 mice by inoculating Aβ_(1-15) vaccine

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Science China(Life Sciences) 2008年第8期51卷 743-750页

作者： HU JinJia1, LI GuoYing2, WANG HuaQiao3, LIN Xian3 & YAO ZhiBin3 1 department of anatomy, Medical college of Shanghai Jiao Tong university, Shanghai 200025, China 2 department of anatomy, Preclinical college, Guangdong university of Pharmacy, Guangzhou 510006, China 3 department of anatomy, Zhongshan School of medicine, Sun Yat-sen university, Guangzhou 510080, China 1. department of anatomy Medical College of Shanghai Jiao Tong University Shanghai 200025 China 2. department of anatomy Preclinical College Guangdong University of Pharmacy Guangzhou 510006 China 3. department of anatomy Zhongshan School of Medicine Sun Yat-sen University Guangzhou 510080 China

This study aims to discuss the effect of preventing pathological changes and cognitive degeneration of Tg2576 mice by inoculating the subunit fragment of Aβ ***-two Tg2576 mice were randomly divided into four groups, each having eight mice:Group I, the control group, inoculated with adjuvants;Group II, the Aβ42 group, inoculated with Aβ42 vaccine;Group Ⅲ, the Aβ1―15 group, inoculated with Aβ1―15 vaccine;and Group IV, the Aβ36―42 group, inoculated with Aβ36―42 *** titer of the serum anti-body against Aβ42(Group II) was significantly higher than that of the control group(Group I), and a low level of antibodies could be detected in the brain homogenate in the three vaccine-inoculated *** water maze test showed that the Aβ42 group, Aβ1―15 group and Aβ36―42 group were obviously im-proved compared with the control *** cultured splenocytes sampled from each group were induced by Con A or their respective antigens, and the cell proliferation of the three vaccine-inoculated groups was significantly higher than that of the control *** the Aβ42 group, IL2 and IFN-γ were relatively low and IL4 and IL10 were relatively *** contrast, IL4 and IL10 were much higher in the Aβ1―15 group and IL2 and IFN-γ were much higher in the Aβ36―42 *** immunohistochemical test showed a large number of senile plaques in the brain cortex and hippocampus of the mice in the con-trol group, no senile plaque in the brain of the Aβ1―15 group and Aβ42 group mice, and a small number of senile plaques in the brain of the Aβ36―42 group *** results suggest that the subunit fragment of Aβ1―15 vaccine could prevent not only cognitive and behavioral degeneration but also Aβ deposition and formation of senile plaques in Tg2576 mice.

关键词： Alzheimer’s disease β-amyloid peptide vaccine Tg2576 mice senile plaque

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The SHP-2 tyrosine phosphatase: Signaling mechanisms and biological functions

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cell Research 2000年第4期10卷 279-288页

作者： QU CHENG KUI department of Hematopoiesis, Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross, Rockville, department of anatomy and cell biology, George Washington university School of medicine and Health Sciences, Washington DC department of Hematopoiesis American Red Cross Rockville MD 20855 USA. quc@***

cellular biological activities are tightly controlled by intracellular signaling processes initiated by extracellular signals. Protein tyrosine phosphatases, which remove phosphate groups from phosphorylated signaling molecules, play equally important tyrosine roles as protein tyrosine kinases in signal transduction. SHP-2, a cytoplajsmic SH2 domain containing protein tyrosine phosphatase, is involved in the signaling pathways of a variety of growth factors and cytokines. Recent studies have clearly demonstrated that this phosphatase plays an important role in transducing signal relay from the cell surface to the nucleus, and is a critical intracellular regulator in mediating cell proliferation and differentiation.

关键词： SHP-2 SHP-1 Signal transduction.?

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Identification of glutamate transporters and receptors in mouse testis

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Acta Pharmacologica Sinica 2004年第03期25卷 112-117页

作者： Jia-hua HU,Na YANG2,Ying-hua MA,Jie JIANG,Jin-fu ZHANG3,Jian FEI4,Li-he GUO4 Laboratory of Molecular cell biology,Institute of Biochemistry and cell biology,Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences,Shanghai 200031 3department of Urology,Tongji Hospital,Tongji university,Shanghai 200065,China Laboratory of Molecular cell biology Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological Sciences Chinese Academy of Sciences department of Urology Tongji Hospital Tongji University

AIM: To investigate the presence of glutamate transporters and receptors in mouse testis. METHODS: Glutamate uptake analysis was performed to study the function of glutamate transporters in mouse testis. Comparative RT-PCR technique and sequencing analysis were used to study the expression of glutamate receptors and transport- ers in mouse testis. RESULTS: Mouse testis possessed glutamate uptake capacity with sodium-dependence. Vmax value of glutamate uptake was (1.60 ± 0.21) pmol/min per mg protein and Km value of glutamate uptake was (11.0±1.6) μmol/L in mouse testis according to saturation analysis. Furthermore, the uptake activity could be inhibited by DHK (GLT1 selective inhibitor) and THA (glutamate uptake inhibitor). In addition, RT-PCR results revealed that glutamate transporters (GLT1 and EAAC1) and ionotropic glutamate receptors (NR1, NR2B, GluR6 and KA2) were expressed in mouse testis. CONCLUSION: Glutamate transporters and receptors do exist in mouse testis.

关键词： glutamates receptors amino acid transport systems testis uptake

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Inhibition of ethanol-induced toxicity by tanshinone ⅡA in PC12 cells

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Acta Pharmacologica Sinica 2006年第6期27卷 659-664页

作者： Xian-fang MENG~2 Xiao-jing ZOU~3 Bin PENG~2 Jing SHI~2 Xin-min GUAN~2 Chun ZHANG~(4,5)2 department of neurobiology,Tongji Medical college,Huazhong university of Science and Technology,Wuhan 430030,China 3 department of Anesthesiology,4 department of Nephrology,Union Hospital,Tongji Medical college,Huazhong university of Science and Technology,Wuhan 430022.China department of neurobiology Tongji Medical College Huazhong University of Science and Technology Wuhan China department of Anesthesiology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China department of Nephrology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:To observe the effects of tanshinone IIA(Tan IIA)on the neurotoxicityinduced by ethanol in PC 12 cells and to explore its protective ***:PC 12cell survival was measured by MTT *** formation of reactive oxygenspecies(ROS)and lactate dehydrogenase(LDH)release were detected by 2′,7′-dichlorofluorescin(DCF)fluorescence and calorimetric method,*** of cell apoptosis was monitored by flow *** expression ofp53 was detected by immuno-fluorescence and flow ***:Ethanolsignificantly impaired the survival of PC12 cells as demonstrated by MTT *** also induced significant ROS formation and increased LDH ***-incubation with Tan IIA in the culture medium significantly reversed these *** caused cell apoptosis and the upregulation of p53 *** anti-apoptosis effects of Tan IIA on ethanol-induced toxicity were accompanied by thedownregulation of pro-apoptotic p53 protein ***:Tan IIAcan protect neurons from apoptosis and might serve as a potential therapeuticdrug for neurological disorders induced by ethanol.

关键词： apoptosis ethanol PC12 cells p53 tanshinone ⅡA

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