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Dynamics of morphological changes for mitochondrial fission and fusion

Science china(Physics,Mechanics & Astronomy) 2010年第4期53卷 680-689页

作者： WANG ShiQi 1,2,4 , FU ChangLiang 1,2 , ZHANG Yan 1,2 , CHEN Quan 3 & LONG Mian 1,2 1 Key Laboratory of Microgravity (National Microgravity Laboratory), beijing 100190, china 2 Center for Biomechanics and Bioengineering, Institute of Mechanics, Chinese Academy of Sciences, beijing 100190, china 3 State Key Laboratory of Biomembrane and Membrane biotechnology, Institute of Zoology, Chinese Academy of Sciences, beijing 100101, china 4 Institute of Vertebrate Paleontology and Paleoanthropology, Chinese Academy of Sciences, beijing 100044, china Key Laboratory of Microgravity (National Microgravity Laboratory) Beijing China Center for Biomechanics and Bioengineering Institute of Mechanics Chinese Academy of Sciences Beijing China Institute of Vertebrate Paleontology and Paleoanthropology Chinese Academy of Sciences Beijing China State Key Laboratory of Biomembrane and Membrane biotechnology Institute of Zoology Chinese Academy of Sciences Beijing China

Mitochondria experience continuous fusion and fission in a living cell, but their dynamics remains poorly quantified. Here a theoretical model was developed, upon a simplified population balance equation (PBE), to predict the morphological changes induced by mitochondrial fission and fusion. Assuming that both fission and fusion events are statistically independent, the survival probability of mitochondria staying in the fission or fusion state was formulated as an exponentially-decayed function with time, which depended on the time-dependent distribution of the mitochondrial volume and the fission and fusion rates. Parametric analysis was done for two typical volume distributions. One was Gamma distribution and the other was Gaussian distribution, derived from the measurements of volume distribution for individual mitochondria in a living cell and purified mitochondria in vitro. The predictions indicated that the survival probability strongly depended on morphological changes of individual mitochondria and was inversely correlated to the fission and fusion rates. This work provided a new insight into quantifying the mitochondrial dynamics via monitoring the evolution of the mitochondrial volume.

关键词： mitochondria morphology fission fusion dynamics modeling

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A Human t-PA Mutant cDNA Cassette Knocked in the Murine fgfr-4 Locus Targeting for Mammary Gland Expression

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Acta Biochimica et Biophysica Sinica 2004年第7期36卷 450-456页

作者： Fu-Yu LIN#, Hong-Xing CHEN#, Xuan CHENG, Xiao YANG*, Ji-Xian DENG*, and Pei-Tang HUANG*beijing Institute of biotechnology, No.20 Dongdajie Street, Fengtai District, beijing 100071, china beijing Institute of biotechnology No.20 Dongdajie StreetFengtai District Beijing 100071 China

The expression of foreign gene in transgenic animals produced by pronuclear microinjection is often confounded by the position effects caused by not only the nature of chromosomal integration site but also the number and arrangement of multiple transgene copies. Gene targeting provides a new way to overcome these inhibitions by introducing single-copy transgene into a chosen site. The choice of a good chromosomal site will favor transgene expression in a predictable fashion. In this study, we tested a new site (fgfr-4) for foreign gene integration and expression. A t-PA mutant (t-PAm) expression cassette under bovine αs1-casein regulatory sequences was efficiently knocked-in fgfr-4 site through homologous recombination. The t-PAm was expressed in the milk of all targeted mice. Our experiment indicates that the fgfr-4 may be a candidate site for knocking foreign gene to make transgenic animals.

关键词： gene targeting fgfr-4 transgenic mammary gland bioreactor integration site t-PA mutant

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A Genome-wide Functional Characterization of Arabidopsis Regulatory Calcium Sensors in Pollen Tubes

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Journal of Integrative Plant Biology 2009年第8期51卷 751-761页

作者： Liming Zhou1,2, Ying Fu1,2 and Zhenbiao Yang2,3 (1State Key Laboratory of Plant Physiology and Biochemistry, Department of Plant Sciences, College of Biological Sciences, china Agricultural University, beijing 100193, china 2china Agricultural University (CAU)-University of California, Riverside (UCR) Joint Center for Biological Sciences and biotechnology, College of Biological Sciences, china Agricultural University, beijing 100193, china 3Center for Plant Cell Biology and Department of Botany and Plant Sciences, University of California, Riverside, CA 92521, USA) State Key Laboratory of Plant Physiology and Biochemistry Department of Plant Sciences College of Biological Sciences China Agricultural University Beijing 100193 China china Agricultural University (CAU)-University of California Riverside (UCR) Joint Center for Biological Sciences and Biotechnology College of Biological Sciences China Agricultural University Beijing 100193 China Center for Plant Cell Biology and Department of Botany and Plant Sciences University of California Riverside CA 92521 USA)

Calcium, an ubiquitous second messenger, plays an essential and versatile role in cellular signaling. The diverse function of calcium signals is achieved by an excess of calcium sensors. Plants possess large numbers of calcium sensors, most of which have not been functionally characterized. To identify physiologically relevant calcium sensors in a specific cell type, we conducted a genome-wide functional survey in pollen tubes, for which spatiotemporal calcium signals are well-characterized and required for polarized tip growth. *** members of calrnodulin （CAM）, CaM-like （CML）, calcium-dependent protein kinase （CDPK） and calcineurin B-like protein （CBL） families were tagged with green fluorescence protein （GFP） and their localization patterns and overexpression phenotypes were characterized in tobacco pollen tubes. We found that several fusion proteins showed distinct overexpression phenotypes and subcellular localization patterns. CDPK24-GFP was localized to the vegetative nucleus and the generative cell/sperms. CDPK32-GFP caused severe growth depolarization. CBL2-GFP and CBL3-GFP exhibited dynamic patterns of subcellular localization, including several endomembrane compartments, the apical plasma membrane （PM）, and cytoskeleton-like structures in pollen tubes. Their overexpression also inhibited pollen tube elongation and induced growth depolarization. These putative calcium sensors are excellent candidates for the calcium sensors responsible for the regulation of calcium homeostasis and calciumdependent tip growth and growth oscillation in pollen tubes.

关键词： Arabidopsis calcium sensor calcium signaling Rho-related GTPase from plants subcellular localization tip growth

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Analysis of the essential DNA region for OsEBP-89 promoter in response to methyl jasmonic acid

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Science china(Life Sciences) 2008年第3期51卷 280-285页

作者： LI Ang1,2, CHEN LiangLiang1,3, REN HaiYun3, WANG XueChen2, ZHANG HaiWen1,4 & HUANG Rong- Feng1,4 1 biotechnology Research Institute, Chinese Academy of Agricultural Sciences, beijing 100081, china 2 National Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, china Agricultural University, beijing 100094, china 3 College of Life Sciences, beijing Normal University, beijing 100875, china 4 National Key Facility of Crop Gene Resources and Genetic Improvement, beijing 100081, china biotechnology Research Institute Chinese Academy of Agricultural Sciences Beijing China National Laboratory of Plant Physiology and Biochemistry College of Biological Sciences China Agricultural University Beijing China College of Life Sciences Beijing Normal University Beijing China National Key Facility of Crop Gene Resources and Genetic Improvement Beijing China

In rice, the characterization of OsEBP-89 is inducible by various stress- or hormone-stimuli, including ethylene, abscisic acid (ABA), jasmonate acid (JA), drought and cold. Here, we report the investigation of essential DNA region within OsEBP-89 promoter for methyl jasmonic acid (MeJA) induction. PLACE analysis indicates that this promoter sequence contains multiple potential elements in response to various stimuli. First, we fused this promoter with GUS gene and analyzed its expression under MeJA treatment through Agrobacterium infiltration mediating transient expression in tobacco leaves. Our results revealed that this chimeric gene could be inducible by MeJA in tobacco leaves. To further de- termine the crucial sequences responsible for MeJA induction, we generated a series of deletion pro- moters which were fused with GUS reporter gene respectively. The results of transient expression of GUS gene driven by these mutant promoters show that the essential region for MeJA induction is po- sitioned in the region between -1200 and -800 in OsEBP-89 promoter containing a G-box (?1127), which is distinct from the essential region containing ERE (?562) for ACC induction. In all, our finding is helpful in understanding the molecular mechanism of OsEBP-89 expression under different stimuli.

关键词： OsEBP-89 essential DNA region methyl jasmonic acid transient assay promoter tobacco leaves

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Functions and mechanisms of the CBL-CIPK signaling system in plant response to abiotic stress

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Progress in Natural Science:Materials International 2009年第6期19卷 667-676页

作者： Ruifen Li a,1, Junwen Zhang a,b,1, Jianhua Wei a, Hongzhi Wanga, Yanzhen Wanga, Rongcai Ma a, a beijing Agro-biotechnology Research Center, beijing Academy of Agriculture and Forestry Sciences, beijing 100097, china b Life Science College, Capital Normal University, beijing 100037, china beijing Agro-biotechnology Research Center Beijing Academy of Agriculture and Forestry Sciences Beijing 100097 China Life Science College Capital Normal University Beijing 100037 China

To cope with environmental stimuli, plants have evolved precise regulatory mechanisms to perceive, transduce and respond to abiotic stresses that can negatively affect growth and development. The CBL-CIPK signaling system is a newly emerging plant-specific and Ca2+-dependent network mediating abiotic stress tolerance. CBLs may sense a Ca2+ signature triggered by abiotic stresses, and have specific interactions with novel CIPK-type kinases after binding Ca2+. The CBL/CIPK complexes may post-translationally phosphory-late downstream target proteins to regulate abiotic stress tolerance in a cell or tissue-specific manner. In some cases transcription factors are induced to activate stress-responsive genes that control adaptation reactions. The CBL-CIPK signaling system exhibits specificity, diversity and complexity. Meanwhile, cross talk also exists in the CBL-CIPK signaling. To date, significant progress has been made in the role of the CBL-CIPK signaling system in responding to salt, low K+ and to high pH, which will provide a fast and efficient method of molecular design breeding combined with the CBL/CIPK engineering of crop plants, for enhanced tolerance to abiotic stresses. However, more CBL/CIPK components remain to be identified, particularly from specific plants that grow in conditions with abiotic stress, and the specificity of their abiotic stress signaling will need to be dissected.

关键词： Calcium signal CBL CIPK Signal transduction Abiotic stress

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Anti-nociceptive role of neuropeptide Y in the nucleus accumbens in rats with inflammation, an effect modulated by mu- and kappa-opioid receptors

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Progress in Natural Science:Materials International 2006年第2期16卷 133-141页

作者： LI Jinju and YU Longchuan (Laboratory of Neurobiology, College of Life Sciences, and National Laboratory of Biomembrane and Membrane biotechnology, Peking University, beijing 100871, china) Laboratory of Neurobiology College of Life Sciences National Laboratory of Biomembrane and Membrane Biotechnology Peking University Beijing 100871 China

Recent study in our laboratory showed that neuropeptide Y (NPY) plays an antinociceptive role in the nucleus accum-bens (NAc) in intact rats. The present study was performed to further investigate the effect of NPY in nociceptive modulation in the NAc of rats with inflammation, and the possible interaction between NPY and the opioid systems. Experimental inflammation was induced by subcutaneous injection of carrageenan into the left hindpaw of rats. Intra-NAc administration of NPY induced a dose-dependent increase of hindpaw withdrawal latencies (HWLs) to thermal and mechanical stimulations in rats with inflammation. The anti-nociceptive effect of NPY was significantly blocked by subsequent intra-NAc injection of the Y1 receptor antagonist NPY28-36, suggesting an involvement of Y1 receptor in the NPY-induced anti-nociception. Furthermore, intra-NAc administration of the opioid antagonist naloxone significantly antagonized the increased HWLs induced by preceding intra-NAc injection of NPY, suggesting an involvement of the endogenous opioid system in the NPY-induced anti-nociception in the NAc during inflammation. Moreover, the NPY-induced anti-nociception was attenuated by following intra-NAc injection of the μ-opioid antagonist β-funaltrexamine (β-FNA), and κ-opioid antagonist nor-binaltorphimine (nor-BNI), but not by S-opioid antagonist naltrindole, indicating that μ- and K-opioid receptors, not S-opioid receptor, are involved in the NPY-induced anti-nociception in the NAc in rats with inflammation.

关键词： nucleus accumbens, neuropeptide Y, Y1 receptor, carrageenan anti-nociception, μ- and κ-opioid receptors.

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Survival of Human Metallothionein-2 Transplastomic Chlamydomonas reinhardtii to Ultraviolet B Exposure

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Acta Biochimica et Biophysica Sinica 2006年第3期38卷 187-193页

作者： Yu-Kun ZHANG~1 Gui-Fang SHEN~2 Bing-Gen RU~1*1 National Laboratory of Protein Engineering and Plant Genetic Engineering,Peking University,beijing 100871,china 2 biotechnology Research Institute,the Chinese Academy of Agricultural Sciences,beijing 100081,china National Laboratory of Protein Engineering and Plant Genetic Engineering Peking UniversityBeijing 100871 China biotechnology Research Institute the Chinese Academy of Agricultural SciencesBeijing 100081 China

Solar ultraviolet (UV) radiation has a great influence on green organisms,especially planktonlike Chlamydomonas.A human metallothionein-2 gene,which is generally considered to have an anti-radiationfunction by its coding product,was transferred into the chloroplast genome of Chlamydomonas *** dynamically measure the UV effects on Chlamydomonas cells grown in liquid tris-acetate-phosphatemedium,a new method was developed based on the relationship between the chlorophyll content of an algalculture and its absorbance at 570 nm after the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium *** this experiment,both the wild-type and the transplastomic *** ceils were cultivated in 96-well microplates containing liquid tris-acetate-phosphate medium in the absence or presence of zinc,copper,cadmium and *** transgenic *** showed a higher resistance than wild-type to UV-Bexposure under all the examined *** in the medium had positive impacts on both types of cells,but had significant influence only on the transplastomic ***,the high cell viability of the trans-genie alga at the end of the 8 h UV-B treatment disappeared after a 20-h recovery *** did notprotect cells from UV-B damage,but clearly enhanced the growth of both wild-type and transgenic ***.

关键词： ultraviolet B MTT transplastomic Chlamydomonas metallothionein

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Expression of two insect-resistant genes cryIA (b&c)/GNA in transgenic tobacco plants results in added protection against both cotton bollworm and aphids

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Chinese Science Bulletin 1999年第22期44卷 2051-2058页

作者： WANG Zhibin and QUO SanduiLaboratory of Biochemistry and Molecular Biology , biotechnology Research Center, Chinese Academy of Agricultural Sciences , beijing 100081, china Laboratory of Biochemistry and Molecular Biology Biotechnology Research Center Chinese Academy of Agricultural Sciences Beijing China

The synthesized Bacillus thuringiensis insecticidal protein gene crylA(b&c) and the synthesized gene GNA, (the mannose specific lectin from snowdrop ( Galanthus nivalis)), tumefaciens have been inserted into plant expression vector pGW4BAI. Leave stripes of Nico-tiana tabacum var. K326 have been transformed with Agrobacterium tumefaciens strain LBA4404 harboring the plant expression vector. 28 kanamycin resistant tobacco plants

关键词： crylA(b&c) snowdrop lectin insect-resistance transgenic plants cotton bollworm aphids pyramiding.

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Assembly and structure of protein phosphatase 2A

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Science china(Life Sciences) 2009年第2期52卷 135-146页

作者： SHI Yi Gong Center for Structural Biology, Department of Biological Sciences and biotechnology, and School of Medicine, Tsinghua University, beijing 100084, china Center for Structural Biology Department of Biological Sciences and Biotechnology and School of Medicine Tsinghua University Beijing China

Protein phosphatase 2A (PP2A) represents a conserved family of important protein serine/threonine phosphatases in species ranging from yeast to human. The PP2A core enzyme comprises a scaffold subunit and a catalytic subunit. The heterotrimeric PP2A holoenzyme consists of the core enzyme and a variable regulatory subunit. The catalytic subunit of PP2A is subject to reversible methylation, medi-ated by two conserved enzymes. Both the PP2A core and holoenzymes are regulated through interac-tion with a large number of cellular cofactors. Recent biochemical and structural investigation reveals critical insights into the assembly and function of the PP2A core enzyme as well as two families of holoenzyme. This review focuses on the molecular mechanisms revealed by these latest advances.

关键词： protein phosphorylation dephosphorylation PP2A structure mechanism

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Electrophysiological research of synapse formation from beginning to maturity

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Chinese Science Bulletin 1998年第1期43卷 55-59页

作者： XIE Zuoping and ZHENG Yufang State Key Laboratory of Biomembrane and Membrane Biotechnique Department of Biological Sciences and biotechnology, Tsinghua University, beijing 100084, china State Key Laboratory of Biomembrane and Membrane Biotechnique Tsinghua University Beijing China Department of Biological Sciences and biotechnology Tsinghua University Beijing China

The development of synapses is an important question in neurobiology. We employ patchclamp technique to record MEPCs and EPCs of myoballs in Xenopus cell culture. The rate of EPC/MEPC is related with synapses maturation. Accordingly, we define the mature synapses as those whose rate of presynaptic membrane’s EPC to MEPC is more than 8. The process of maturation needs about 2-5 h after neurites contact with myoballs. We divide the development of synapses into three stages: young, developing and mature. The neurotransmitters stack gradually and increase by hundreds of times along with young synapses growing into maturity. So the process of synapses maturation should include presynaptic membrane specialization, postsynaptic membrane specialization and neurotransmitters stack.

关键词： definition of mature synapse α latrotoxin (α LTX) neurotransmitter stack.

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